U.S. patent application number 13/253536 was filed with the patent office on 2012-04-05 for methods and compounds for the targeted delivery of agents to bone for interaction therewith.
This patent application is currently assigned to Pradama, Inc.. Invention is credited to Kevyn Merten, William M. Pierce, JR., K. Grant Taylor, Leonard C. Waite.
Application Number | 20120083440 13/253536 |
Document ID | / |
Family ID | 39710536 |
Filed Date | 2012-04-05 |
United States Patent
Application |
20120083440 |
Kind Code |
A1 |
Pierce, JR.; William M. ; et
al. |
April 5, 2012 |
METHODS AND COMPOUNDS FOR THE TARGETED DELIVERY OF AGENTS TO BONE
FOR INTERACTION THEREWITH
Abstract
Bone targeted compounds and methods are provided. Compounds can
include a Bone Targeting Portion (R.sub.T), having an affinity for
bone; a Bone Active Portion (R.sub.A) for interacting with and
affecting bone; and a Linking Portion (R.sub.L) connecting the Bone
Targeting Portion and the Bone Active Portion.
Inventors: |
Pierce, JR.; William M.;
(Louisville, KY) ; Taylor; K. Grant; (Louisville,
KY) ; Waite; Leonard C.; (Corydon, IN) ;
Merten; Kevyn; (Louisville, KY) |
Assignee: |
Pradama, Inc.
University of Louisville Research Foundation, Inc.
|
Family ID: |
39710536 |
Appl. No.: |
13/253536 |
Filed: |
October 5, 2011 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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12036057 |
Feb 22, 2008 |
8071575 |
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13253536 |
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60891299 |
Feb 23, 2007 |
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60893375 |
Mar 7, 2007 |
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Current U.S.
Class: |
514/2.3 ;
514/178; 514/182; 514/252.19; 514/34; 514/363; 514/456; 514/616;
530/322; 536/6.4; 544/295; 548/139; 549/403; 552/626; 552/638;
564/153; 564/157 |
Current CPC
Class: |
C07D 311/36 20130101;
C07C 2603/40 20170501; A61P 19/08 20180101; A61P 21/06 20180101;
A61P 35/00 20180101; C07C 2603/44 20170501; C07D 285/125 20130101;
C07C 237/44 20130101; A61P 19/00 20180101; A61P 19/10 20180101;
A61K 31/70 20130101; A61P 31/00 20180101; A61P 31/04 20180101 |
Class at
Publication: |
514/2.3 ;
564/157; 514/616; 552/626; 514/182; 549/403; 514/456; 564/153;
552/638; 514/178; 548/139; 514/363; 536/6.4; 514/34; 544/295;
514/252.19; 530/322 |
International
Class: |
A61K 31/167 20060101
A61K031/167; C07J 41/00 20060101 C07J041/00; A61K 31/567 20060101
A61K031/567; C07D 311/34 20060101 C07D311/34; A61K 31/353 20060101
A61K031/353; A61K 31/569 20060101 A61K031/569; C07D 285/125
20060101 C07D285/125; A61K 31/433 20060101 A61K031/433; C07H 15/252
20060101 C07H015/252; A61K 31/704 20060101 A61K031/704; C07D 417/14
20060101 C07D417/14; A61K 31/506 20060101 A61K031/506; C07K 9/00
20060101 C07K009/00; A61K 38/14 20060101 A61K038/14; A61P 19/00
20060101 A61P019/00; A61P 19/10 20060101 A61P019/10; A61P 35/00
20060101 A61P035/00; A61P 31/00 20060101 A61P031/00; C07C 237/40
20060101 C07C237/40 |
Claims
1. A compound of the formula: a) ##STR00319## b) ##STR00320## or c)
##STR00321## or pharmaceutically acceptable salts or solvates
thereof, wherein R.sub.T is ##STR00322## wherein R.sub.T is
connected to the compound at R.sub.1, R.sub.2, or R.sub.4; wherein
R.sub.1 is hydrogen, lower alkyl, alkyl, aryl lower alkyl, aryl, or
a covalent bond when R.sub.T is connected to the compound at
R.sub.1; R.sub.2 is hydrogen, lower alkyl, alkyl, aryl lower alkyl,
aryl, or a covalent bond when R.sub.T is connected to the compound
at R.sub.2; R.sub.3 is hydrogen, lower alkyl, alkyl, aryl lower
alkyl, aryl, or carbonyl-containing; R.sub.4 is hydrogen, lower
alkyl, alkyl, aryl lower alkyl, aryl, carbonyl-containing, or a
covalent bond when R.sub.T is connected to the compound at R.sub.4;
R.sub.5 and R.sub.6 are independently hydrogen, lower alkyl, or
alkyl, or R.sub.5 and R.sub.6, taken together with the carbon atoms
to which they are bonded, form a ring containing about 6 to about
14 ring carbon atoms and up to a total of about 18 carbon atoms,
which formed ring can be monocyclic, bicyclic, or tricyclic,
wherein the ring can optionally have substituents, including
heteroatoms; R.sub.7 is hydroxy, lower alkoxy, or NR.sub.8,
R.sub.9; R.sub.8 and R.sub.9 are independently hydrogen, or lower
alkyl; wherein i is 0-3, k is 0-3, and p is 1-4; wherein each
R.sub.q is independently hydrogen or hydroxy; wherein X is O, NH,
S, or covalent bond; wherein X' is O, NH, S, or covalent bond;
wherein m is 1-3, n is 1-4, and when m>1, each n is
independently 1-4; wherein each R.sub.S is independently hydrogen,
hydroxy, lower alkyl, or lower alkyl with heteroatoms; wherein D
and G are independently covalent bond, carbonyl, epoxy, or
anhydride; wherein E is covalent bond, (CT.sub.2).sub.r, where T is
hydrogen, hydroxy, or lower alkyl, and where r is 0-8, or
(C).sub.r, where r is 2-8, and where the carbons are unsaturated or
partially saturated with hydrogen; so long as n is 2-4 when D, E,
and G are all covalent bond, X is NH or O, and R.sub.S is H, or D
is carbonyl, E and G are covalent bond, and R.sub.S is H; and
wherein R.sub.A is hydrogen, hydroxyl, a protecting group, or a
Bone Active Portion derived from a bone active agent.
2. The compound of claim 1, wherein R.sub.5 and R.sub.6 are
hydrogen.
3. The compound of claim 1, wherein R.sub.7 is NR.sub.8R.sub.9.
4. The compound of claim 3, wherein R.sub.8 and R.sub.9 are both
hydrogen.
5. The compound of claim 1, wherein R.sub.3 is hydrogen.
6. The compound of claim 1, wherein R.sub.T is ##STR00323## and
R.sub.T is connected to the compound at R.sub.1.
7. The compound of claim 1, wherein R.sub.T is ##STR00324## and
R.sub.T is connected to the compound at R.sub.1.
8. The compound of claim 1, wherein R.sub.T is ##STR00325## and
R.sub.T is connected to the compound at R.sub.1.
9. The compound of claim 1, according to the formula ##STR00326##
where n' and n'' are independently 1-4, and X'' is O, NH, S, or
covalent bond.
10. The compound of claim 1, according to the formula
##STR00327##
11. The compound of claim 1, according to the formula
##STR00328##
12. The compound of claim 1, according to the formula
##STR00329##
13. The compound of claim 12, wherein R.sub.T is: ##STR00330## and
R.sub.T is connected to the compound at R.sub.1 or R.sub.4.
14. The compound of claim 13, according to the formula
##STR00331##
15. The compound of claim 14, wherein the compound is provided as a
salt.
16. The compound of claim 15, wherein the salt is a chloride
salt.
17. The compound of claim 1, according to the formula
##STR00332##
18. The compound of claim 17, wherein R.sub.T is: ##STR00333## and
R.sub.T is connected to the compound at R.sub.1 or R.sub.4.
19. The compound of claim 1, according to the formula
##STR00334##
20. The compound of claim 19, wherein R.sub.A is hydrogen or
hydroxyl.
21. The compound of claim 19, wherein R.sub.A is a protecting
group.
22. The compound of claim 19, wherein R.sub.A is a bone active
portion derived from a bone active agent selected from the bone
active agents set forth in Tables A-D.
23. The compound of claim 19, wherein R.sub.A is a bone active
agent selected from a steroid, a non-steroidal estrogenic agent, a
nitric oxide agent, an androgen, a carbonic anhydrase inhibitor, an
anti-cancer agent, and an antimicrobial agent.
24. The compound of claim 19, wherein the bone active agent is an
anti-cancer agent.
25. The compound of claim 19, wherein the bone active agent is a
tyrosine kinase inhibitor.
26. The compound of claim 1, according to the formula
##STR00335##
27. The compound of claim 1, according to the formula
##STR00336##
28. The compound of claim 1, according to the formula ##STR00337##
wherein E is CH.sub.2 or (CH.sub.2).sub.2.
29. The compound of claim 1, according to the formula
##STR00338##
30. The compound of claim 1, according to the formula ##STR00339##
wherein E is CH.sub.2 or (CH.sub.2).sub.2.
31. The compound of claim 1, according to the formula
##STR00340##
32. The compound of claim 31, wherein R.sub.A is hydrogen or
hydroxyl.
33. The compound of claim 31, wherein R.sub.A is a protecting
group.
34. The compound of claim 31, wherein R.sub.A is a bone active
portion derived from a bone active agent selected from the bone
active agents set forth in Tables A-D.
35. The compound of claim 34, wherein R.sub.A is a bone active
portion derived from a steroid.
36. The compound of claim 34, wherein R.sub.A is a bone active
portion derived from an estrogenic agent.
37. The compound of claim 36, wherein R.sub.A is a bone active
portion derived from a steroidal estrogenic agent.
38. The compound of claim 37, where the steroidal estrogenic agent
is estradiol.
39. The compound of claim 36, wherein R.sub.A is a bone active
portion derived from a non-steroidal estrogenic agent.
40. The compound of claim 39, wherein the non-steroidal estrogenic
agent is genistein.
41. The compound of claim 31, wherein R.sub.A is a bone active
portion derived from a nitric oxide agent.
42. The compound of claim 41, wherein the nitric oxide agent is
alkoxy-(NO.sub.2).sub.2.
43. The compound of claim 31, wherein R.sub.A is a bone active
portion derived from an androgen.
44. The compound of claim 43, wherein the androgen is DHEA.
45. The compound of claim 43, wherein the androgen is
Testosterone.
46. The compound of claim 31, wherein R.sub.A is a bone active
portion derived from a carbonic anhydrase inhibitor
47. The compound of claim 46, wherein R.sub.A is a bone active
portion derived from 2-amino-1,3,4-thiadiazole-5-sulfonamide.
48. The compound of claim 31, wherein R.sub.A is a bone active
portion derived from an anti-cancer agent.
49. The compound of claim 48, wherein the anti-cancer agent is
doxorubicin.
50. The compound of claim 31, wherein R.sub.A is a tyrosine kinase
inhibitor.
51. The compound of claim 50, wherein the tyrosine kinase inhibitor
is dasatinib.
52. The compound of claim 31, wherein R.sub.A is a bone active
portion derived from an antimicrobial agent
53. The compound of claim 52, wherein the antimicrobial agent is
vancomycin.
54. A method for treating a bone condition in a subject in need
thereof, comprising: administering to the subject an effective
amount of the compound of claim 1.
55. The method of claim 54, wherein the bone condition is a
metabolic bone disease.
56. The method of claim 55, wherein the metabolic bone disease is
osteoporosis, and wherein R.sub.A is a bone active portion derived
from a bone active agent selected from: an androgen, a steroidal
estrogenic agent, a non-steroidal estrogenic agent, a nitric oxide
agent, and a carbonic anhydrase inhibitor.
57. The method of claim 56, wherein the subject has a primary
condition associated with osteoporosis.
58. The method of claim 56, wherein administration of the compound
has an anabolic effect on the bone of the subject.
59. The method of claim 54, wherein the bone condition is a primary
or a secondary bone cancer, and wherein R.sub.A is a bone active
portion derived from an anti-cancer agent.
60. The method of claim 59, wherein the bone condition is a
secondary bone cancer.
61. The method of claim 54, wherein the subject has a primary
cancer associated with a secondary bone cancer.
62. The method of claim 61, wherein the primary cancer is breast,
lung, prostate, kidney, thyroid cancer, or multiple myeloma.
63. The method of claim 54, wherein the bone condition is a
microbial infection, and wherein R.sub.A is a bone active portion
derived from an antimicrobial agent.
64. The method of claim 63, wherein the bone condition is
osteomyelitis, and wherein R.sub.A is a bone active portion derived
from an antimicrobial agent.
65. The method of claim 64, wherein the osteomyelitis is associated
with a prosthetic joint infection.
66. The method of claim 54, wherein the subject has a primary
infection associated with osteomyelitis.
Description
RELATED APPLICATIONS
[0001] This application is a continuation of U.S. patent
application Ser. No. 12/036,057 filed Feb. 22, 2008, which claims
priority from U.S. Provisional Application Ser. No. 60/891,299
filed Feb. 23, 2007, and U.S. Provisional Application Ser. No.
60/893,375 filed Mar. 7, 2007, the entire disclosures of which are
incorporated herein by this reference.
TECHNICAL FIELD
[0002] The presently-disclosed subject matter relates to prevention
and treatment of bone disorders and conditions, and, more
particularly, to the targeted delivery of prophylactic and
therapeutic agents to bone.
BACKGROUND
[0003] Bone is a dynamic tissue, consisting of cells in a protein
matrix, upon which is superimposed a crystalline structure of
various calcium salts. Because bone is the primary structural
support system for the body of an animal, bone disorders can create
substantial problems. Bone disorders include, for example,
fractures, suboptimal mechanical competence, suboptimal bone blood
perfusion, suboptimal bone healing ability, cancerous
transformation (primary and bone metastasis), and infection.
[0004] Bone disorders can occur in a variety of manners. For
example, bone disorders can result from excessive forces being
exerted onto the bone, primary bone conditions, and secondary bone
conditions associated with other conditions. Bone conditions
include, for example, metabolic bone diseases (MBDs). MBDs are
conditions characterized by weakening of bones, which weakening is
associated with suboptimal mechanical competence and an increased
likelihood of fracturing. Osteoporosis is an example of a MBD.
Osteoporosis is characterized by bone degeneration caused by a
relative excess of bone resorption. Clinical osteoporosis is found
in approximately 25% of postmenopausal women, and subclinical
osteoporosis, which is responsible for untold numbers of bone
fractures, is far more widespread. Other examples of MBDs include,
but are not limited to: Paget's disease, characterized by an
abnormal growth of bone such that the bone is larger and weaker
than normal bone; and osteogenesis imperfecta, characterized by
bones that are abnormally brittle.
[0005] In addition to serving as a rigid support for the body of an
animal, bone is an organ that responds to various agents. To the
extent that bone has the ability to interact with and respond to
certain agents, disorders associated with bone conditions can be
prevented, diagnosed, or treated using appropriate agents having
the ability to interact with and affect a desired response in bone.
For example, with regard to osteoporosis, there are certain agents,
which are thought to interact with bone and are currently available
for the treatment or prevention of the condition. Such agents
include: bisphosphonates (e.g., alendronate, risedronate);
calcitonin; selective estrogen receptor modulators (SERMs) (e.g.,
raloxifene); selective androgen receptor modulators (SARMs); growth
factors; cytokines; agents used for estrogen or hormone replacement
therapy (ET/HRT); and parathyroid hormone (PTH) (e.g.,
teriparatide).
[0006] There are a variety of disadvantages associated with
treatment using these known agents. For example, although PTH has
some anabolic activity, biphosphonates, calcitonin, SERMs, and
ET/RHT are primarily anti-catabolic, operating to limit bone
resportion. In this regard, the anti-catabolic compounds only treat
osteoporosis in so much as they attempt to keep bone density from
further decreasing. There are also various side effects associated
with such agents; for example, bisphosphonate treatment is
associated with gastrointestinal and esophageal erosion, and has
been implicated in osteonecrosis of the jaw; SERM treatment has
been associated with deep vein thrombosis and hot flashes; ET/HRT
has been implicated in increased risk of breast cancer and
cardiovascular disease; and PTH therapy has been suggested to
potentially increase risk of osteosarcoma (osteogenic sarcoma), a
type of cancer that develops in bone, is characterized by formation
of a bone matrix having decreased strength relative to normal
non-malignant bone matrix, and which can metastasize to other bones
and other organs. See e.g., Bilezikian J P (2006) N Engl J Med
355:2278-2281; Cranney A, Adachi J D (2005) Drug Saf 28:721-730;
Marshall J K (2002) Expert Opin Drug Saf 1:71-78; Rossouw J E, et
al., Writing Group for the Women's Health Initiative Investigators
(2002) Risks and benefits of estrogen plus progestin in healthy
postmenopausal women: principal results From the Women's Health
Initiative randomized controlled trial. JAMA 288:321-333; Vahle J
L, et al. (2002) Toxicol Pathol 30:312-321, which are incorporated
herein by this reference. There are also various drawbacks
associated with the delivery of such known agents to an animal, for
example, bisphosphonates demonstrate poor oral bioavailability,
calcitonin is not orally deliverable, and PTH must be injected.
Additionally, some known agents have a limited capacity to affect
bone because they lack a specific affinity for bone. That is to say
that, when some of the known agents are delivered to an animal,
they are not specifically directed to the bone. In this regard,
when some of the known agents are delivered to an animal, they are
delivered to non-specific locations in the body of the animal, such
that they fail to interact with the bone or require a large dose to
affect a response in bone. Also in this regard, when such agents
are delivered to an animal, they can be directed to undesirable
locations in the body of the animal, resulting in undesirable side
effects.
[0007] Accordingly, there remains a need in the art for compounds,
systems, and methods for treating bone disorders and conditions
that satisfactorily address some or all of the above-identified
disadvantages.
SUMMARY
[0008] The presently-disclosed subject matter meets some or all of
the above-identified needs, as will become evident to those of
ordinary skill in the art after a study of information provided in
this document.
[0009] This Summary describes several embodiments of the
presently-disclosed subject matter, and in many cases lists
variations and permutations of these embodiments. This Summary is
merely exemplary of the numerous and varied embodiments. Mention of
one or more representative features of a given embodiment is
likewise exemplary. Such an embodiment can typically exist with or
without the feature(s) mentioned; likewise, those features can be
applied to other embodiments of the presently-disclosed subject
matter, whether listed in this Summary or not. To avoid excessive
repetition, this Summary does not list or suggest all possible
combinations of such features.
[0010] The presently-disclosed subject matter includes compounds,
or pharmaceutically acceptable compositions thereof, having an
affinity for bone, or "bone targeted compounds." The
presently-disclosed subject matter includes bone targeted compounds
and methods useful for treating conditions of interest, e.g.,
conditions affecting bone. The presently-disclosed subject matter
further includes methods for delivering an agent of interest to
bone.
[0011] The bone targeted compounds of the presently-disclosed
subject matter can in some embodiments generally include three
units. The three units of the compounds are: a Bone Targeting
Portion (R.sub.T), having an affinity for bone; a Linking Portion
(R.sub.L) that is capable of connecting the Bone Targeting Portion
to a third unit; and the third unit (R.sub.A). As such, the
compounds of the presently-disclosed subject matter can be
represented by the following formula:
##STR00001##
[0012] The third unit (R.sub.A) can be a Bone Active Portion, a
protecting group, or hydrogen or hydroxyl. In embodiments where
R.sub.A is a Bone Active Portion, the Bone Active Portion interacts
with and affects the bone. The Bone Active Portion can be derived
from a Bone Active Agent, which can be selected for its efficacy in
treating a condition of interest. In embodiments where R.sub.A is a
protecting group, the protecting group can assist with maintaining
the stability of the compound, for example, by keeping an adjacent
group on the linking portion from reacting with other portions of
the compound, e.g., cyclizing. Compounds including a protecting
group can be stably stored until it becomes desirable to associate
the compound with a Bone Active Portion derived from a Bone Active
Agent of interest. In this regard, the compounds including a
protecting group are useful for preparing compounds for treating
conditions associated with bone. In some embodiments, R.sub.A is
hydrogen or hydroxyl, depending on the embodiment of the Linking
Portion, R.sub.L, being used, as will be described below. The
presently-disclosed subject matter includes salts derived from
compounds where R.sub.A is hydrogen or hydroxyl, which salts can be
stably stored until it becomes desirable to associate the compound
with a Bone Active Portion. In this regard, the compounds in which
R.sub.A is hydrogen or hydroxyl are useful for preparing compounds
for treating conditions associated with bone.
[0013] In some embodiments, the compound of the presently-disclosed
subject matter can be represented by the formula:
##STR00002##
where R.sub.T is
##STR00003##
and where R.sub.T is connected to the compound at R.sub.1, R.sub.2,
or R.sub.4.
[0014] In some embodiments, R.sub.1 can be hydrogen, lower alkyl,
alkyl, aryl lower alkyl, aryl, or a covalent bond when R.sub.T is
connected to the compound at R.sub.1. R.sub.2 can be hydrogen,
lower alkyl, alkyl, aryl lower alkyl, aryl, or a covalent bond when
R.sub.T is connected to the compound at R.sub.2. R.sub.3 can be
hydrogen, lower alkyl, alkyl, aryl lower alkyl, aryl, or
carbonyl-containing R.sub.4 can be hydrogen, lower alkyl, alkyl,
aryl lower alkyl, aryl, carbonyl-containing, or a covalent bond
when R.sub.T is connected to the compound at R.sub.4. R.sub.5 and
R.sub.6 can be independently hydrogen, lower alkyl, or alkyl, or
R.sub.5 and R.sub.6, taken together with the carbon atoms to which
they are bonded, can form a ring containing about 6 to about 14
ring carbon atoms and up to a total of about 18 carbon atoms, which
formed ring can be monocyclic, bicyclic, or tricyclic, wherein the
ring can optionally have substituents, including heteroatoms.
R.sub.7 can be hydroxy, lower alkoxy, or NR.sub.8, R.sub.9; and
R.sub.8 and R.sub.9 can be independently hydrogen, or lower alkyl.
In some embodiments of the compounds, i can be 0, 1, 2, or 3, k can
be 0, 1, 2, or 3, p can be 0, 1, 2, 3, or 4, and each R.sub.q is
independently hydrogen or hydroxyl. In some embodiments, X can be
O, NH, S, or a covalent bond, and X' can be O, NH, S, or a covalent
bond. In some embodiments of the compounds, m can be 1, 2, or 3, n
can be 1, 2, 3, or 4, and when m>1, each n is independently 1,
2, 3, or 4; each R.sub.S can independently be hydrogen, hydroxy,
lower alkyl, or lower alkyl with heteroatoms; D and G can be
independently a covalent bond, carbonyl, epoxy, or anhydride; and E
can be a covalent bond, (CT.sub.2).sub.r, where T is hydrogen,
hydroxy, or lower alkyl, and where r is 0, 1, 2, 3, 4, 5, 6, 7, or
8, or (C).sub.r, where r is 2, 3, 4, 5, 6, 7, or 8, and where the
carbons are unsaturated or partially saturated with hydrogen.
R.sub.A can be hydrogen, hydroxyl, a protecting group, or a Bone
Active Portion.
[0015] In some embodiments, R.sub.5 and R.sub.6 are hydrogen. In
some embodiments, R.sub.7 is NR.sub.9R.sub.9. In some embodiments,
R.sub.8 and R.sub.9 are both hydrogen. In some embodiments, R.sub.3
is hydrogen.
[0016] In some embodiments, R.sub.T is
##STR00004##
and R.sub.T is connected to the compound at R.sub.1. In some
embodiments, R.sub.T is
##STR00005##
and R.sub.T is connected to the compound at R.sub.1. In some
embodiments, R.sub.T is
##STR00006##
and R.sub.T is connected to the compound at R.sub.1. In some
embodiments, R.sub.T is
##STR00007##
and R.sub.T is connected to the compound at R.sub.4.
[0017] In some embodiments, the compounds of the
presently-disclosed subject matter can be represented by the
formula
##STR00008##
where n' and n'' are independently 1, 2, 3, or 4, and X'' is O, NH,
S, or a covalent bond.
[0018] In some embodiments, the compounds of the
presently-disclosed subject matter can be represented by the
formula
##STR00009##
In some embodiments, the compounds of the presently-disclosed
subject matter can be represented by the formula
##STR00010##
In some embodiments, the compounds of the presently-disclosed
subject matter can be represented by the formula
##STR00011##
In some embodiments, the compounds of the presently-disclosed
subject matter can be represented by the formula
##STR00012##
In some embodiments, the compounds of the presently-disclosed
subject matter can be represented by the formula
##STR00013##
In some embodiments, the compounds of the presently-disclosed
subject matter can be represented by the formula
##STR00014##
[0019] In some embodiments, the compounds of the
presently-disclosed subject matter can be represented by the
formula
##STR00015##
[0020] In some embodiments, the compounds of the
presently-disclosed subject matter can be represented by the
formula
##STR00016##
[0021] In some embodiments, the compounds of the
presently-disclosed subject matter can be represented by the
formula
##STR00017##
In some embodiments, R.sub.A is hydrogen. In some embodiments,
wherein R.sub.A is hydrogen, the compound can be represented by the
formula
##STR00018##
In some embodiments, the compound is provided as a salt. In some
embodiments, the salt is a chloride salt. In some embodiments,
R.sub.A is a protecting group. In some embodiments, R.sub.A is a
bone active portion derived from a bone active agent selected from
the bone active agents set forth in Tables A-D. In some
embodiments, R.sub.A is a bone active portion derived from a
steroid. In some embodiments, R.sub.A is a bone active portion
derived from an estrogenic agent. In some embodiments, R.sub.A is a
bone active portion derived from a steroidal estrogenic agent. In
some embodiments, the steroidal estrogenic agent is estradiol. In
some embodiments, the compound can be represented by the
formula
##STR00019##
In some embodiments, the compound can be represented by the
formula
##STR00020##
In some embodiments, the compound can be represented by the
formula
##STR00021##
In some embodiments, the compound can be represented by the
formula
##STR00022##
In some embodiments, the compound can be represented by the
formula
##STR00023##
In some embodiments, R.sub.A is a bone active portion derived from
a non-steroidal estrogenic agent. In some embodiments, the
non-steroidal estrogenic agent is genistein. In some embodiments,
the compound can be represented by the formula
##STR00024##
In some embodiments, the compound can be represented by the
formula
##STR00025##
In some embodiments, R.sub.A is a bone active portion derived from
a nitric oxide agent. In some embodiments, the nitric oxide agent
is alkoxy-(NO.sub.2).sub.2. In some embodiments, the compound can
be represented by the formula
##STR00026##
In some embodiments, the compound can be represented by the
formula
##STR00027##
In some embodiments, R.sub.A is a bone active portion derived from
an androgen. In some embodiments, androgen is testosterone. In some
embodiments, the androgen is DHEA. In some embodiments, the
compound can be represented by the formula
##STR00028##
In some embodiments, the compound can be represented by the
formula
##STR00029##
In some embodiments, R.sub.A is a bone active portion derived from
a carbonic anhydrase inhibitor. In some embodiments, R.sub.A is a
bone active portion derived from a
2-amino-1,3,4-thiadiazole-5-sulfonamide. In some embodiments,
R.sub.A is a bone active portion derived from an anti-cancer agent.
In some embodiments, the anti-cancer agent is doxorubicin. In some
embodiments, the compound can be represented by the formula
##STR00030##
In some embodiments, the compound can be represented by the
formula
##STR00031##
In some embodiments, the anti-cancer agent is dasatinib. In some
embodiments, the compound can be represented by the formula
##STR00032##
In some embodiments, the compound can be represented by the
formula
##STR00033##
In some embodiments, the compound can be represented by the
formula
##STR00034##
wherein E can be CH.sub.2 or (CH.sub.2).sub.2. In some embodiments,
R.sub.A is a bone active portion derived from an antimicrobial
agent. In some embodiments, the antimicrobial agent is vancomycin.
In some embodiments, the compound can be represented by the
formula
##STR00035##
In some embodiments, the compound can be represented by the
formula
##STR00036##
In some embodiments, the compounds of the presently-disclosed
subject matter can be represented by the formula
##STR00037##
In some embodiments, R.sub.A is hydroxyl. In some embodiments,
R.sub.A is a protecting group. In some embodiments, R.sub.A is a
bone active portion derived from a bone active agent selected from
the bone active agents set forth in Tables A-D. In some
embodiments, R.sub.A is a bone active portion derived from a
steroid. In some embodiments, R.sub.A is a bone active portion
derived from an estrogenic agent. In some embodiments, R.sub.A is a
bone active portion derived from a steroidal estrogenic agent. In
some embodiments, the steroidal estrogenic agent is estradiol. In
some embodiments, R.sub.A is a bone active portion derived from a
non-steroidal estrogenic agent. In some embodiments, the
non-steroidal estrogenic agent is genistein. In some embodiments,
R.sub.A is a bone active portion derived from a nitric oxide agent.
In some embodiments, the nitric oxide agent is
alkoxy-(NO.sub.2).sub.2. In some embodiments, R.sub.A is a bone
active portion derived from an androgen. In some embodiments, the
androgen is DHEA. In some embodiments, the androgen is
Testosterone. In some embodiments, the compound can be represented
by the formula
##STR00038##
In some embodiments, the compound can be represented by the
formula
##STR00039##
In some embodiments, R.sub.A is a bone active portion derived from
a carbonic anhydrase inhibitor. In some embodiments, R.sub.A is a
bone active portion derived from
2-amino-1,3,4-thiadiazole-5-sulfonamide. In some embodiments, the
compound can be represented by the formula
##STR00040##
In some embodiments, the compound can be represented by the
formula
##STR00041##
In some embodiments, R.sub.A is a bone active portion derived from
an anti-cancer agent. In some embodiments, the anti-cancer agent is
doxorubicin. In some embodiments, the anti-cancer agent is
dasatinib. In some embodiments, R.sub.A is a bone active portion
derived from an antimicrobial agent. In some embodiments, the
antimicrobial agent is vancomycin.
[0022] In some embodiments, the method for treating a bone
condition in a subject in need thereof includes, administering to
the subject an effective amount of a compound of the
presently-disclosed subject matter. In some embodiments, the bone
condition is a metabolic bone disease. In some embodiments, the
metabolic bone disease is osteoporosis, and R.sub.A is a bone
active portion derived from a bone active agent selected from: an
androgen, a steroidal estrogenic agent, a non-steroidal estrogenic
agent, a nitric-oxide-targeted/generating agent, and a carbonic
anhydrase inhibitor. In some embodiments, the subject has a primary
condition associated with osteoporosis. In some embodiments,
administration of the compound has an anabolic effect on the bone
of the subject. In some embodiments, the bone condition is a
primary or a secondary bone cancer, and wherein R.sub.A is a bone
active portion derived from an anti-cancer agent. In some
embodiments, the bone condition is a secondary bone cancer. In some
embodiments, the subject has a primary cancer associated with a
secondary bone cancer. In some embodiments, the primary cancer is
breast, lung, prostate, kidney, or thyroid cancer. In some
embodiments, the bone condition is a microbial infection, and
wherein R.sub.A is a bone active portion derived from an
antimicrobial agent. In some embodiments, the bone condition is
osteomyelitis, and R.sub.A is a bone active portion derived from an
antimicrobial agent. In some embodiments, the osteomyelitis is
associated with a prosthetic joint infection. In some embodiments,
the subject has a primary infection associated with
osteomyelitis.
BRIEF DESCRIPTION OF THE DRAWINGS
[0023] FIG. 1 is a line graph depicting body weight as a function
of time for animals administered 17-ethinyl estradiol, alendronate,
parathyroid hormone, the compound of Formula 175 (BTA-2), or the
compound of Formula 176 (BTA-3).
[0024] FIG. 2 is a bar graph depicting the uterine mass of animals
administered 17-ethinyl estradiol, alendronate, parathyroid
hormone, the compound of Formula 175 (BTA-2), or the compound of
Formula 176 (BTA-3).
[0025] FIG. 3 is a bar graph depicting the ratio of uterine mass to
body weight of animals administered 17-ethinyl estradiol,
alendronate, parathyroid hormone, the compound of Formula 175
(BTA-2), or the compound of Formula 176 (BTA-3).
[0026] FIG. 4 is a bar graph depicting the whole bone density of
animals administered 17-ethinyl estradiol, alendronate, parathyroid
hormone, the compound of Formula 175 (BTA-2), or the compound of
Formula 176 (BTA-3).
[0027] FIG. 5 is a bar graph depicting the regional bone density of
the proximal left femur of animals administered 17-ethinyl
estradiol, alendronate, parathyroid hormone, the compound of
Formula 175 (BTA-2), or the compound of Formula 176 (BTA-3).
[0028] FIG. 6 is a bar graph depicting the regional bone density of
the distal left femur of animals administered 17-ethinyl estradiol,
alendronate, parathyroid hormone, the compound of Formula 175
(BTA-2), or the compound of Formula 176 (BTA-3).
[0029] FIG. 7 is a line graph depicting body weight as a function
of time for animals administered 17-ethinyl estradiol, alendronate,
parathyroid hormone, or the compound of Formula 131 (BTE2-pg2-D2)
at doses of 10, 100, or 1000 .mu.g/kg.
[0030] FIG. 8 is a bar graph depicting the uterine mass of animals
administered 17-ethinyl estradiol, alendronate, parathyroid
hormone, or the compound of Formula 131 (BTE2-pg2-D2) at doses of
10, 100, or 1000 .mu.g/kg.
[0031] FIG. 9 is a bar graph depicting the ratio of uterine mass to
body weight of animals administered 17-ethinyl estradiol,
alendronate, parathyroid hormone, or the compound of Formula 131
(BTE2-pg2-D2) at doses of 10, 100, or 1000 .mu.g/kg.
[0032] FIG. 10 is a bar graph depicting the whole bone density of
animals administered 17-ethinyl estradiol, alendronate, parathyroid
hormone, or the compound of Formula 131 (BTE2-pg2-D2) at doses of
10, 100, or 1000 .mu.g/kg.
[0033] FIG. 11 is a bar graph depicting the regional bone density
of the proximal left femur of animals administered 17-ethinyl
estradiol, alendronate, parathyroid hormone, or the compound of
Formula 131 (BTE2-pg2-D2) at doses of 10, 100, or 1000
.mu.g/kg.
[0034] FIG. 12 is a bar graph depicting the regional bone density
of the distal left femur of animals administered 17-ethinyl
estradiol, alendronate, parathyroid hormone, or the compound of
Formula 131 (BTE2-pg2-D2) at doses of 10, 100, or 1000
.mu.g/kg.
[0035] FIG. 13 is a bar graph depicting the regional bone density
of the left femoral diaphysis of animals administered 17-ethinyl
estradiol, alendronate, parathyroid hormone, or the compound of
Formula 131 (BTE2-pg2-D2) at doses of 10, 100, or 1000
.mu.g/kg.
[0036] FIG. 14 is a bar graph depicting serum osteocalcin levels
for animals administered 17-ethinyl estradiol, alendronate,
parathyroid hormone, or the compound of Formula 131 (BTE2-pg2-D2)
at doses of 100 or 1000 .mu.g/kg.
[0037] FIG. 15 is a line graph depicting body weight as a function
of time for animals administered 17-ethinyl estradiol, alendronate,
parathyroid hormone, or the compound of Formula 132 (BTE2-pg3-D2)
at doses of 10, 100, or 1000 .mu.g/kg.
[0038] FIG. 16 is a bar graph depicting the uterine mass of animals
administered 17-ethinyl estradiol, alendronate, parathyroid
hormone, or the compound of Formula 132 (BTE2-pg3-D2) at doses of
10, 100, or 1000 .mu.g/kg.
[0039] FIG. 17 is a bar graph depicting the ratio of uterine mass
to body weight of animals administered 17-ethinyl estradiol,
alendronate, parathyroid hormone, or the compound of Formula 132
(BTE2-pg3-D2) at doses of 10, 100, or 1000 .mu.g/kg.
[0040] FIG. 18 is a bar graph depicting the whole bone density of
animals administered 17-ethinyl estradiol, alendronate, parathyroid
hormone, or the compound of Formula 132 (BTE2-pg3-D2) at doses of
10, 100, or 1000 .mu.g/kg.
[0041] FIG. 19 is a bar graph depicting the regional bone density
of the proximal left femur of animals administered 17-ethinyl
estradiol, alendronate, parathyroid hormone, or the compound of
Formula 132 (BTE2-pg3-D2) at doses of 10, 100, or 1000
.mu.g/kg.
[0042] FIG. 20 is a bar graph depicting the regional bone density
of the distal left femur of animals administered 17-ethinyl
estradiol, alendronate, parathyroid hormone, or the compound of
Formula 132 (BTE2-pg3-D2) at doses of 10, 100, or 1000
.mu.g/kg.
[0043] FIG. 21 is a bar graph depicting the regional bone density
of the left femoral diaphysis of animals administered 17-ethinyl
estradiol, alendronate, parathyroid hormone, or the compound of
Formula 132 (BTE2-pg3-D2) at doses of 10, 100, or 1000
.mu.g/kg.
[0044] FIG. 22 is a bar graph illustrating trabecular volume
fraction data for animals administered 17-ethinyl estradiol,
alendronate, parathyroid hormone, or the compound of Formula 132
(BTE2-pg3-D2) at doses of 10, 100, or 1000 .mu.g/kg.
[0045] FIG. 23 includes three-dimensional images of bone that were
constructed using data collected by a customized micro-CT system,
for animals administered 17-ethinyl estradiol, alendronate,
parathyroid hormone, or the compound of Formula 132 (BTE2-pg3-D2)
at doses of 10, 100, or 1000 .mu.g/kg.
[0046] FIG. 24 is a bar graph illustrating tibial bone strength of
animals administered 17-ethinyl estradiol, alendronate, parathyroid
hormone, or the compound of Formula 132 (BTE2-pg3-D2) at doses of
10, 100, or 1000 .mu.g/kg.
[0047] FIG. 25 is a line graph depicting body weight as a function
of time for animals administered 17-ethinyl estradiol, alendronate,
parathyroid hormone, or the compound of Formula 133 (BTE2-pg2-D3)
at doses of 10, 100, or 1000 .mu.g/kg.
[0048] FIG. 26 is a bar graph depicting the uterine mass of animals
administered 17-ethinyl estradiol, alendronate, parathyroid
hormone, or the compound of Formula 133 (BTE2-pg2-D3) at doses of
10, 100, or 1000 .mu.g/kg.
[0049] FIG. 27 is a bar graph depicting the ratio of uterine mass
to body weight of animals administered 17-ethinyl estradiol,
alendronate, parathyroid hormone, or the compound of Formula 133
(BTE2-pg2-D3) at doses of 10, 100, or 1000 .mu.g/kg.
[0050] FIG. 28 is a bar graph depicting the regional bone density
of animals administered 17-ethinyl estradiol, alendronate,
parathyroid hormone, or the compound of Formula 133 (BTE2-pg2-D3)
at doses of 10, 100, or 1000 .mu.g/kg.
[0051] FIG. 29 is a bar graph depicting the regional bone density
of the proximal left femur of animals administered 17-ethinyl
estradiol, alendronate, parathyroid hormone, or the compound of
Formula 133 (BTE2-pg2-D3) at doses of 10, 100, or 1000
.mu.g/kg.
[0052] FIG. 30 is a bar graph depicting the regional bone density
of the distal left femur of animals administered 17-ethinyl
estradiol, alendronate, parathyroid hormone, or the compound of
Formula 133 (BTE2-pg2-D3) at doses of 10, 100, or 1000
.mu.g/kg.
[0053] FIG. 31 is a bar graph depicting the regional bone density
of the left femoral diaphysis of animals administered 17-ethinyl
estradiol, alendronate, parathyroid hormone, or the compound of
Formula 133 (BTE2-pg2-D3) at doses of 10, 100, or 1000
.mu.g/kg.
[0054] FIG. 32 is a bar graph illustrating trabecular volume
fraction data for animals administered 17-ethinyl estradiol,
alendronate, parathyroid hormone, or the compound of Formula 133
(BTE2-pg2-D3) at doses of 10, 100, or 1000 .mu.g/kg.
[0055] FIG. 33 includes three-dimensional images of bone that were
constructed using data collected by a customized micro-CT system,
for animals administered 17-ethinyl estradiol, alendronate,
parathyroid hormone, or the compound of Formula 133 (BTE2-pg2-D3)
at doses of 10, 100, or 1000 .mu.g/kg.
[0056] FIG. 34 is a bar graph depicting bone resorption or
osteoclast-mediated breakdown of collagen type I in bone by
measuring the C-telopeptide fragment of collagen type I (CTX-I) in
animals administered 17-ethinyl estradiol, alendronate, parathyroid
hormone, or the compound of Formula 133 (BTE2-pg2-D3) at a dose of
1000 .mu.g/kg.
[0057] FIG. 35 is a line graph depicting body weight as a function
of time for animals administered 17-ethinyl estradiol, alendronate,
parathyroid hormone, or the compound of Formula 141
(BT-Testosterone) at a dose of 3 mg/kg.
[0058] FIG. 36 is a bar graph depicting the uterine mass of animals
administered 17-ethinyl estradiol, alendronate, parathyroid
hormone, or the compound of Formula 141 (BT-Testosterone) at a dose
of 3 mg/kg.
[0059] FIG. 37 is a bar graph depicting the ratio of uterine mass
to body weight of animals administered 17-ethinyl estradiol,
alendronate, parathyroid hormone, or the compound of Formula 141
(BT-Testosterone) at a dose of 3 mg/kg.
[0060] FIG. 38 is a bar graph depicting the whole bone density of
animals administered 17-ethinyl estradiol, alendronate, parathyroid
hormone, or the compound of Formula 141 (BT-Testosterone) at a dose
of 3 mg/kg.
[0061] FIG. 39 is a bar graph depicting the regional bone density
of the proximal left femur of animals administered 17-ethinyl
estradiol, alendronate, parathyroid hormone, or the compound of
Formula 141 (BT-Testosterone) at a dose of 3 mg/kg.
[0062] FIG. 40 is a bar graph depicting the regional bone density
of the distal left femur of animals administered 17-ethinyl
estradiol, alendronate, parathyroid hormone, or the compound of
Formula 141 (BT-Testosterone) at a dose of 3 mg/kg.
[0063] FIG. 41A includes photographs of a colony formation assay
for cancer cells treated with increasing concentrations of the
compound of Formula 146 (BT2-pg2-doxorubicin (Nbr-VI)) or
increasing concentrations of doxorubicin.
[0064] FIG. 41B is a bar graph depicting the results of a colony
formation assay for cancer cells treated with increasing
concentrations of the compound of Formula 146 (BT2-pg2-doxorubicin
(Nbr-VI)) or increasing concentrations of doxorubicin.
[0065] FIG. 42 is a bar graph illustrating cell proliferation data
for day 5 and day 6 for cells treated with increasing
concentrations of doxorubicin.
[0066] FIG. 43 is a bar graph illustrating cell proliferation data
for day 5 and day 6 for cells treated with increasing
concentrations of the compound of Formula 146 (BT2-pg2-doxorubicin
(Nbr-VI)).
[0067] FIG. 44 is a bar graph illustrating cell proliferation data
for day 1 for cells treated with increasing concentrations of the
compound of Formula 146 (BT2-pg2-doxorubicin (Nbr-VI)) or
increasing concentrations of doxorubicin.
[0068] FIG. 45 is a bar graph illustrating cell proliferation data
for day 2 for cells treated with increasing concentrations of the
compound of Formula 146 (BT2-pg2-doxorubicin (Nbr-VI)) or
increasing concentrations of doxorubicin.
[0069] FIG. 46 is a bar graph illustrating cell proliferation data
for day 3 for cells treated with increasing concentrations of the
compound of Formula 146 (BT2-pg2-doxorubicin (Nbr-VI)) or
increasing concentrations of doxorubicin.
[0070] FIG. 47 is a bar graph illustrating cell proliferation data
for day 4 for cells treated with increasing concentrations of the
compound of Formula 146 (BT2-pg2-doxorubicin (Nbr-VI)) or
increasing concentrations of doxorubicin.
[0071] FIG. 48 is a bar graph illustrating cell proliferation data
for day 5 for cells treated with increasing concentrations of the
compound of Formula 146 (BT2-pg2-doxorubicin (Nbr-VI)) or
increasing concentrations of doxorubicin.
[0072] FIG. 49 is a bar graph illustrating cell proliferation data
for day 6 for cells treated with increasing concentrations of the
compound of Formula 146 (BT2-pg2-doxorubicin (Nbr-VI)) or
increasing concentrations of doxorubicin.
DESCRIPTION OF EXEMPLARY EMBODIMENTS
[0073] The details of one or more embodiments of the
presently-disclosed subject matter are set forth in this document.
Modifications to embodiments described herein, and other
embodiments, will be evident to those of ordinary skill in the art
after a study of the information provided in this document. The
information provided herein, and particularly the specific details
of the described exemplary embodiments, is provided primarily for
clearness of understanding and no unnecessary limitations are to be
understood therefrom. In case of conflict, the specification of
this document, including definitions, will control.
[0074] Unless defined otherwise, all technical and scientific terms
used herein have the same meaning as commonly understood by one of
ordinary skill in the art to which the presently-disclosed subject
matter belongs. Although any methods, devices, and materials
similar or equivalent to those described herein can be used in the
practice or testing of the presently-disclosed subject matter,
representative methods, devices, and materials are now
described.
[0075] Following long-standing patent law convention, the terms
"a," "an," and "the" refer to "one or more" when used in this
application, including the claims. Thus, for example, reference to
"a cell" includes a plurality of such cells, and so forth.
[0076] Unless otherwise indicated, all numbers expressing
quantities of ingredients, properties such as reaction conditions,
and so forth used in the specification and claims are to be
understood as being modified in all instances by the term "about".
Accordingly, unless indicated to the contrary, the numerical
parameters set forth in this specification and claims are
approximations that can vary depending upon the desired properties
sought to be obtained by the presently-disclosed subject
matter.
[0077] As used herein, the term "about," when referring to a value
or to an amount of mass, weight, time, volume, concentration or
percentage is meant to encompass variations of in some embodiments
.+-.20%, in some embodiments .+-.10%, in some embodiments .+-.5%,
in some embodiments .+-.1%, in some embodiments .+-.0.5%, and in
some embodiments .+-.0.1% from the specified amount, as such
variations are appropriate to perform the disclosed method.
[0078] As used in the present specification, the following words
and phrases are generally intended to have the meanings as set
forth below, except to the extent that the context in which they
are used indicates otherwise.
[0079] "Lower alkyl," refers to alkyl groups with the general
formula C.sub.nH.sub.2n+1, where n=1 to about 6. In some
embodiments, n=1 to about 3. The groups can be straight-chained or
branched. Examples include methyl, ethyl, propyl, isopropyl,
n-butyl, sec-butyl, t-butyl, isobutyl, n-pentyl, isopentyl,
neopentyl, n-hexyl, and the like.
[0080] "Lower alkyl with heteroatoms," refers to groups with the
general formula C.sub.nX.sub.mH.sub.r, where X is a heteroatom, and
n+m=2 to about 6. In some embodiments, n+m=2 to about 3. The
heteroatom can be selected from: nitrogen, oxygen, sulfur,
phosphorus, boron, chlorine, bromine, iodine, and other
heteroatoms. In some embodiments, the heteroatom is selected from:
nitrogen, oxygen, and sulfur. r=a positive whole number (integer)
that is appropriate in light of n, X, and m, as will be understood
by one of ordinary skill in the art. For example, if n=2, X is
nitrogen, and m=1, then r=6, such that the group is C.sub.2H.sub.6.
The groups can be straight-chained or branched.
[0081] "Alkyl," refers to alkyl groups with the general formula
C.sub.nH.sub.2n+1, where n=about 6 to about 18. The groups can be
straight-chained or branched.
[0082] "Alkyl with heteroatoms," when used alone or in combination
with other groups, refers to groups with the general formula
C.sub.nX.sub.mH.sub.r, where X is a heteroatom, and n+m=about 6 to
about 18. The heteroatom can be selected from: nitrogen, oxygen,
sulfur, phosphorus, boron, chlorine, bromine, iodine, and other
heteroatoms. In some embodiments, the heteroatom is selected from:
nitrogen, oxygen, and sulfur. r=a positive whole number (integer)
that is appropriate in light of n, X, and m, as will be understood
by one of ordinary skill in the art. For example, if n=5, X is
nitrogen, and m=2, then r=13, such that the group is
C.sub.5N.sub.2H.sub.13. The groups can be straight-chained or
branched.
[0083] "Carbonyl-containing," refers to a group containing a
carbonyl, for example, an aldehyde, a ketone, an ester, an amide, a
carboxylic acid, or an acyl group. The groups can include 1 to
about 6 carbon atoms, and at least one oxygen atom.
[0084] "Aryl," refers to an aromatic group containing ring carbon
atoms and having about 5 to about 14 ring carbon atoms and up to a
total of about 18 ring or pendant carbon atoms. Examples include
phenyl, .alpha.-naphthyl, .beta.-naphthyl, tolyl, xylyl, and the
like.
[0085] "Aryl lower alkyl" refers to an aryl group bonded to a
bridging lower alkyl group, as defined herein. Examples include
benzyl, phenethyl, naphthylethyl, and the like.
[0086] Each of the aforementioned groups could be substituted or
unsubstituted. For example, "alkyl" can include substituted alkyl,
substituted with hydroxyl, heteroatoms, or lower alkyl groups.
[0087] The presently-disclosed subject matter includes compounds,
or pharmaceutically acceptable compositions thereof, having an
affinity for bone, or "bone targeted compounds." The
presently-disclosed subject matter includes bone targeted compounds
and methods useful for treating conditions of interest, e.g.,
conditions affecting bone. The presently-disclosed subject matter
further includes methods for delivering an agent of interest to
bone.
[0088] The bone targeted compounds of the presently-disclosed
subject matter can in some embodiments generally include three
units. The three units of the compounds are: a Bone Targeting
Portion, having an affinity for bone; a Linking Portion that is
capable of connecting the Bone Targeting Portion to a third unit;
and the third unit. In some embodiments, the third unit is a Bone
Active Portion, capable of interacting with bone. For example, the
Bone Active Portion could be derived from a Bone Active Agent
having an effect on bone. In other embodiments, the third unit is a
protecting group that assists with maintaining the stability of the
bone targeted compound. In other embodiments, the third unit is a
hydrogen or a hydroxyl group. In some embodiments, the compound is
a salt, derived from a compound wherein the third unit is hydrogen
or hydroxyl, which salt can be maintained stably. Appropriate salts
can be selected, for example, from the Handbook of Pharmaceutical
Salts (Stahl P H, Wermuth C G, editors. 2002. Handbook of
pharmaceutical salts: Properties, selection and use.
Weinheim/Zurich: Wiley-VCH/VHCA), Haynes, et al. 2005. J.
Pharmaceutical Sciences 94, 10:2111-2120, or references cited
thereby, which are incorporated herein by this reference. Examples
of salts include, but are not limited to: ammonium, bromide,
chloride, ethylenediamine, maleate, phosphate, sulfate, tartrate,
and tosylate salts.
[0089] The compounds of the presently-disclosed subject matter can
be represented by the following formula:
##STR00042##
where, R.sub.T represents the Bone Targeting Portion, R.sub.L,
represents the linking portion, and R.sub.A represents the Bone
Active Portion, the protecting group, or the hydrogen or hydroxyl.
In some embodiments, the compounds can be provided as a salt or
solvate, e.g., a pharmaceutically acceptable salt or solvate.
Bone Targeting Portion
[0090] The Bone Targeting Portion (R.sub.T) of the compound has an
affinity for the extracellular inorganic matrix of bone. The Bone
Targeting Portion can be represented by the following formula:
##STR00043##
wherein
[0091] R.sub.1 is hydrogen, lower alkyl, alkyl, aryl lower alkyl,
or aryl;
[0092] R.sub.2 is hydrogen, lower alkyl, alkyl, aryl lower alkyl,
or aryl;
[0093] R.sub.3 is hydrogen, lower alkyl, alkyl, aryl lower alkyl,
aryl, or carbonyl-containing;
[0094] R.sub.4 is hydrogen, lower alkyl, alkyl, aryl lower alkyl,
aryl, or carbonyl-containing;
[0095] R.sub.5 and R.sub.6 are independently hydrogen, lower alkyl,
or alkyl, or R.sub.5 and R.sub.6, taken together with the carbon
atoms to which they are bonded, form a ring containing about 6 to
about 14 ring carbon atoms and up to a total of about 18 carbon
atoms, which formed ring can be monocyclic, bicyclic, or tricyclic,
wherein the ring can optionally have substituents, including
heteroatoms;
[0096] R.sub.7 is hydroxy, lower alkoxy, or NR.sub.8, R.sub.9
and
[0097] R.sub.8 and R.sub.9 are independently hydrogen, or lower
alkyl.
[0098] An exemplary Bone Targeting Portion of the
presently-disclosed subject matter can be represented by the
following formula:
##STR00044##
where R.sub.1, R.sub.3, R.sub.5, and R.sub.6 are each hydrogen;
R.sub.4 is methyl; and R.sub.7 is amino.
[0099] Another exemplary Bone Targeting Portion of the
presently-disclosed subject matter can be represented by the
following formula:
##STR00045##
where R.sub.1, R.sub.3, R.sub.4, R.sub.5, and R.sub.6 are each
hydrogen; and R.sub.7 is amino.
[0100] Another exemplary Bone Targeting Portion of the
presently-disclosed subject matter can be represented by the
following formula:
##STR00046##
[0101] where R.sub.1, R.sub.3, R.sub.5, and R.sub.6 are each
hydrogen; R.sub.4 is benzyl; and R.sub.7 is amino.
[0102] Another exemplary Bone Targeting Portion of the
presently-disclosed subject matter can be represented by the
following formula:
##STR00047##
where R.sub.1, R.sub.2, R.sub.3, R.sub.5, and R.sub.6 are each
hydrogen; and R.sub.7 is amino.
[0103] The linking portion is attached to the Bone Targeting
Portion at R.sub.1, R.sub.2, or R.sub.4. For example, when the
linking portion is attached to the Bone Targeting Portion at
R.sub.1, the compound has the following formula:
##STR00048##
[0104] As another non-limiting example, when the linking portion is
attached to the Bone Targeting Portion at R.sub.7, the compound has
the following formula:
##STR00049##
Bone Active Portion (R.sub.A)
[0105] In some embodiments of the compounds of the
presently-disclosed subject matter, R.sub.A is a Bone Active
Portion of the compound. The Bone Active Portion interacts with and
affects the bone. The Bone Active Portion can be derived from a
Bone Active Agent, which can be selected for its efficacy in
treating a condition of interest. A Bone Active Portion that is
derived from a Bone Active Agent can be modified relative to the
Bone Active Agent as is necessary to be connected to the remainder
of the compound, while maintaining some or all of the activity
associated with the Bone Active Agent, or while obtaining enhanced
activity relative to the Bone Active Agent. For example, a Bone
Active Portion derived from a Bone Active Agent after being linked
to the compound can have the structure of the Bone Active Agent,
less a leaving group (e.g., less a hydrogen, less a hydroxyl, less
a covalent bond, or less another leaving group) or including a
connecting group, as will be apparent to one of ordinary skill in
the art.
[0106] Without wishing to be bound by theory or mechanism, once
embodiments of the compound are delivered to bone for interaction
therewith, the Bone Active Portion could be cleaved from the
compound, becoming a free Bone Active Agent capable of interacting
with adjacent bone. Alternatively, once embodiments of the compound
are delivered to bone, the Bone Active Portion, as part of the
bone-targeted compound, could interact with bone.
[0107] Exemplary Bone Active Portions of the compounds of the
presently-disclosed subject matter can be derived from Bone Active
Agents, including but not limited to steroids, including but not
limited to androgens, steroidal estrogenic agents, and other sex
hormones; estrogenic agents, including but not limited to steroidal
estrogenic agents, estrogen precursors, estrogen analogues and
metabolites, non-steroidal estrogenic agents, including
plant-derived estrogens; carbonic anhydrase inhibitors; nitric
oxide agents; antineoplastic or anti-cancer agents; antimicrobial
agents; and other Bone Active Agents. Examples of Bone Active
Agents from which the Bone Active Portion of the compounds of the
presently-disclosed subject matter can be derived are set forth in
Tables A-D.
TABLE-US-00001 TABLE A Examples of Steroids from which the Bone
Active Portion can be Derived Bone Active Agent (BAA) adrogens,
including but not limited estrogens, including but not limited to
the following: to the following testosterone estradiol
dehydropiandrosterone (DHEA) estrone 5.alpha.-dihydrotestosterone
estriol androstenedione estrogen precursors etiocholanolone
estrogen analogues and metabolites epiandrosterone tibolone
androsterone 2-Methoxyestradiol (2-ME) 17 .alpha.-methyl
testosterone fluoxymesterone 17 .alpha.-ethyl testosterone 17
.alpha.-methylandrostan-3.beta., 17 .beta.-diol androstan-3a, 17
.beta.-diol androstan-3 .alpha.-17 .alpha.-diol androstan-17
.beta.-ol 3-one androstane-17 .alpha.-ol-3 one D5-androsten-3
.alpha., 17 .beta.-diol D5-androstene-3.beta., 17.beta.-diol
androstane-3-17-dione D4-androstenedione Selective androgen
receptor modulators (SARMs)
TABLE-US-00002 TABLE B Examples of Estrogenic Agents from which the
Bone Active Portion can be Derived Bone Active Agent (BAA)
steroidal estrogenic agents, including non-steroidal estrogenic
agents, but not limited to the following: including but not limited
to the estradiol following: estrone genistein estriol resveratrol
estrogen precursors daidzein estrogen analogues and metabolites
glycitein tibolone formononetin 2-Methoxyestradiol (2-ME) biochanin
A diethylstilbestrol hexestrol xenoestrogens phytoestrogens &
mycoestrogens coumestans isoflavonoids ipriflavone lignan
phytoestrogens (including but not limited to: secoisolariciresinol
diglycoside)
TABLE-US-00003 TABLE C Examples of other Bone Active Portions from
which the Bone Active Portion can be Derived Bone Active Agent
(BAA) prostaglandins, including but not carbonic anhydrase
inhibitors, includ- limited to the following ing but not limited to
the following prostaglandin D2 azetazolamide prostaglandin EP4
agonist ONO- 2-amino-1,3,4-thiadiazole-5-sulfon- 4819 amide
prostaglandin E2 6-hydroxy-2-benzothiazole sulfon- prostaglandin E1
amide prostaglandin F2a 6-ethylsuccinyloxy-2-benzothiazole
15-methyl-PGE2 sulfonamide 15-methyl-11-deoxy-PGE1
succinylazolamide cyclooxygenase products derived oxaloylazolamide
from eicosatetraeneoic (22:4) acid etholazolamide and the
corresponding 22:5 and methazolamide 22:6 analogs benzolamide
carbonic anhydrase inhibitors (e.g., as described in U.S. Pat. Nos.
5,641,762; 5,242,937; 5,055,480; and 5,059,613, which are
incorporated herein by this reference) parathyroid hormones,
including cathepsin K Inhibitors, including but but not limited to
the following: not limited to the following: intact PTH (PTH
[1-84]) OST-4077 [furan-2-carboxylic acid (1- teriparatide
(recombinant human {1-[4-fluoro-2-(2-oxo-pyrrolidin-1-yl)- PTH
[1-34]) phenyl]-3-oxo-piperidin-4- PTH fragment (1-31)
ylcarbamoyl}-cyclohexyl)-amide] PTH-related protein (PTHrP) thyroid
hormones, including but nitric oxide agents, including but not not
limited to the following: limited to the following: thyroxine (T4)
nitroglycerine liothyronin (T3) isosorbide mononitrate erythrityl
tetranitriate alkoxy-(NO.sub.2).sub.2 HMG CoA reductase inhibitors,
vitamin D molecules, including but not including but not limited to
the limited to the floowing: following: ergocalciferol, (vitamin
D2) lovastatin cholecalciferol, (vitamin D3) compactin
25-hydroxy-ergocalciferol simvastatin 1,25-dihydroxyergocalciferol
pevastatin 25-hydroxy-cholecalciferol mevastatin
1,25-dihydroxycholecalciferol cerivastatin 24,25-dihydroxy vitamin
D3 fluvastatin pitavastatin atorvastatin selective estrogen
receptor matrix metalloproteinase (MMP) modulators (SERMS),
including inhibitors, including but not limited but not limited to
the following: to the following: raloxifene batimastat arzoxifene
marimastat lasofoxifene prinomastat bazedoxiene tanomastat
droloxifene trocade ispemifene interleukin-6 Receptor Antagonists,
toremifene including but not limited to the tamoxifene floowing:
ormeloxifene 20S,21-epoxy-resibufogenin-3-formate esterns (ERBF)
estren-.alpha. (4-estren-3 .alpha.,17.beta.-diol) integrin
alphavbeta3 inhibitors, estren-.beta.
(4-estren-3.beta.,17.beta.-diol) including but not limited to the
following: 3,4-dichloro-phenylbiguanide
3,5-dichloro-phenylbiguanide Tyrosine kinase inhibitors, CB1- and
CB2-selective cannabinoid including but not limited to the receptor
antagonists following: calcium-Sensing Receptor Antagonists AP23451
chloride channel inhibitors 2-(4-aminocyclohexyl)-9-ethyl-
non-steroidal anti-inflammatory agents N-phenyl-9H-purin-6-amine
(NSAIDs) (See Boyce, et al., Clin. Cancer growth Factors Res. 12,
6291s-6925s (2006), strontium ranelate which is incorporated herein
isotaxiresinol by this reference (N-(5-Chloro- clomiphene
1,3-benzodioxol-4-yl)-7- reveromycin A [2-(4-methylpiperazin-1-
autocoids yl)ethoxy]-5-(tetrahydro- RANK-L antagonists
2H-pyran-4-yloxy)quina- thiazides zolin-4-amine) (a/k/a AZD0530,
ferulic acid See Estell, et al., J. Clinical Oncology, 23, 16S, p.
3041 (2005) (Abstract); and Hennequin, et al., J. Medicinal
Chemistry 49, 22, 6465- 6488 (2006) ,which is incorporated herein
by this reference) [N-(2- chloro-6-methylphenyl)-2-(6-
(4-(2-hydroxyethyl)piperazin- 1-yl)-2-methylpyrimidin-4-
ylamino)thiazole-5-carboxamide (dasatinib) beta-blockers femarelle
(ST56a)
TABLE-US-00004 TABLE D Examples of other Bone Active Portions from
which the Bone Active Portion can be Derived Bone Active Agent
(BAA) anti-caner agents, including but not limited to the follwing:
doxorubicin methoxtrexate cyclophosphamide/cytoxan capecitabine
isosfamide carboplatin vincristine 5-fluorouracil cisplatin
epirubicin etoposide topotecan methotrexate irinotecan taxanes,
e.g., docetaxel, paclitaxel dasatinib vinorelbine erlotinib
gemcitabine gefitinib capecitabine bexarotene carboplatin
vinblastine free radical scavengers antimicrobial agents, including
but not limited to the following: vancomycin
[0108] In some embodiments, anti-cancer agent refers not only to
bone active agents that directly affect cancers cells, but also to
bone active agents that indirectly affect cancer cells, for
example, bone active agents that target the microenvironment of the
cancer cells. In a specific nonlimiting example, the bone active
agent could act upon osteoclasts or osteoblasts, thereby affecting
cancer cell growth, e.g., a RANK-L antagonist could be used as the
bone active agent.
[0109] As noted above, the Bone Active Portion of some embodiments
of the compounds of the presently-disclosed subject matter
interacts with and affect bone, having a desired effect on bone.
The desired effect can vary, for example, based on the bone
condition being treated. Compounds of the presently-disclosed
subject matter can affect bone to treat a variety of bone
conditions, including those set forth in Tables E and F.
TABLE-US-00005 TABLE E Primary Bone Conditions Category (ies) of
Primary Bone Condition Bone Active Agent(s) Metabolic Osteoporosis
Anabolic Agent and/or Bone Diseases Paget's Disease Anti-catabolic
Agent (MBD) Osteogenesis imperfecta Primary hyperparathyroidism
Fibrous dysplacia (McCune-Albright syndrome) Osteopetrosis
Tumor-induced osteomalacia Rickets (nutritional, genetic,
drug-induced) Renal osteodystrophy Fanconi syndrome
Hypophosphatasia Fracture Fracture resulting from a MBD, another
disease or Anabolic Agent disorder, or an external physical force
Cancer Primery bone cancer (e.g., primary bone sarcoma) Anti-cancer
agents
TABLE-US-00006 TABLE F Primary Conditions, with which another
Secondary Bone Conditions is Associated Secondary Category (ies)
Bone Bone Active Primary Condition Condition Agent(s) Alcoholism
Osteoporosis Anabolic Anorexia Nervosa (and other eating disorders)
Agent and/or Asthma, certain treatment programs for; and
Anti-catabolic bone loss associated with rheumatoid arthritis Agent
Autoimmune Diseases, e.g., lupus Celiac Disease (Gluten allergy)
Diabetes Inflammatory Bowel Disease (Crohn's Disease, ulcerative
colitis) Cancer (other than primary bone cancer, Bone Anti-cancer
including: breast, lung, prostate, kidney, metastasis agents
thyroid, multiple myeloma, and other cancers) Infection
Osteomyelitis Antimicrobial agents
[0110] With reference to Table E, in some embodiments, compounds
wherein the Bone Active Portion is derived from a Bone Active Agent
set forth in Tables A-C can be used to treat bone conditions
including metabolic bone diseases. In some embodiments, when a
metabolic bone disease is being treated, an anti-catabolic effect,
an anabolic effect, or a combination thereof is desired. In some
embodiments, compounds wherein the Bone Active Portion is derived
from a Bone Active Agent set forth in Tables A-C can be used to
treat bone conditions including bone fracture. In some embodiments,
when a bone fracture is being treated, an anabolic effect is
desired. In some embodiments, compounds wherein the Bone Active
Portion is derived from a Bone Active Agent that is a nitric oxide
agent (e.g., a nitric oxide agent as set forth in Table C) can be
used to treat bone conditions including bone fracture. In some
embodiments, compounds wherein the Bone Active Portion is derived
from a Bone Active Agent that is a vasodilator can be used to treat
bone conditions including bone fracture.
[0111] With continued reference to Table E, in some embodiments,
compounds wherein the Bone Active Portion is derived from a Bone
Active Agent that is an anti-cancer agent (e.g., anti-cancer agent
set forth in Table D) can be used to treat a primary or a
metastatic bone cancer, where an anti-cancer effect is desired. In
some embodiments, compounds wherein the Bone Active Portion is
derived from a Bone Active Agent that is an anti-microbial agent
can be used to treat a bone infection. For example, compounds
wherein the Bone Active Portion is derived from an antimicrobial
agent (e.g., antimicrobial agent set forth in Table D) can be used
to treat osteomyelitis, where an antimicrobial effect is
desired.
[0112] With reference to Table F, it can sometimes be desirable to
administer to a subject having a primary condition a compound
useful for treating a secondary condition. In some embodiments, a
subject can be identified as having one or more primary conditions
associated with a secondary bone condition that is a metabolic bone
disease, such as osteoporosis, as identified in Table F. The
subject can then be administered a compound for treating
osteoporosis. In some embodiments, such a treatment includes a
prophylactic treatment, e.g., arresting or preventing the
development of osteoporosis. In some embodiments, an anti-catabolic
effect and/or an anabolic effect is desired. In some embodiments, a
subject can be identified as having a primary cancer capable of
metastasizing to bone. The subject can then be administered a
compound for treating bone cancer, wherein an anti-cancer effect is
desired. In this regard, in some embodiments, such a treatment
includes a prophylactic treatment, e.g., arresting or preventing
the development of bone cancer. In some embodiments, a subject can
be identified as having a primary infection capable of leading to a
secondary bone condition, osteomyelitis. The subject can then be
administered a compound for treating osteomyelitis, wherein an
antimicrobial effect is desired. In this regard, in some
embodiments, such a treatment includes a prophylactic treatment,
e.g., arresting or preventing the development of osteomyelitis. In
some embodiments, the osteomyelitis is associated with a prosthetic
joint infection.
[0113] In some embodiments, the compounds including Bone Active
Portions of the presently-disclosed subject matter can have an
anti-catabolic effect on bone. In some embodiments, the compounds
including Bone Active Portions of the presently-disclosed subject
matter can have an anabolic effect on bone. In some embodiments,
the compounds including Bone Active Portions of the
presently-disclosed subject matter can have an anti-catabolic
effect and an anabolic effect on bone. In some embodiments, the
compounds including Bone Active Portions of the presently-disclosed
subject matter can have an anti-cancer effect on bone. In some
embodiments, the compounds including Bone Active Portions of the
presently-disclosed subject matter can have an anti-microbial
effect on bone. In some embodiments, the compounds including Bone
Active Portions of the presently-disclosed subject matter can have
an anti-biotic effect on bone. In some embodiments, the compounds
can be provided in synergistic compositions containing other
compounds useful for treating a primary and/or secondary bone
condition.
[0114] As used herein, a catabolic effect is an effect that results
in a net reduction in bone mass, bone density, and/or bone
strength. As used herein, an anti-catabolic effect is an effect
that results in a decrease in the magnitude of a catabolic effect.
Reduction in bone mass, density, and/or strength can be identified
by comparing a first bone measurement (e.g., control or earlier
time), to a second bone measurement (e.g., treated or later time).
Bone mass, density, and strength can be measured using methods
known to those skilled in the art.
[0115] As used herein, an anabolic effect is an effect that results
in increased bone strength; or increased bone mass or density, and
increased bone strength. Increases bone mass or density, and
increases in bone strength provide evidence that net bone formation
is being promoted. Increases in bone mass or density, and increases
in bone strength can be measured by comparing a first bone
measurement (e.g., control or earlier time), to a second bone
measurement (e.g., treated or later time.) Bone mass or density can
be measured using methods known to those skilled in the art. In
some embodiments, requisite increased bone mass or density affected
by treatment with a compound of interest is an increase of at least
about 5%, at least about 10%, at least about 15%, at least about
20%, at least about 25%, at least about 30%, or at least about 35%,
when a first bone measurement and a second bone measurement are
compared.
[0116] Increased bone strength can be measured by comparing a first
bone strength measurement to a second bone strength measurement. In
some embodiments, the increased bone strength can be measured by
comparing the bone strength of an untreated control (first bone
strength measurement), to the bone strength of a bone sample after
treatment with a compound of interest (second bone strength
measurement). In some embodiments, increased bone strength can be
measured by comparing the bone strength of a bone sample before
treatment with a compound of interest (first bone strength
measurement), to the bone strength of a bone sample after treatment
with the compound of interest (second bone strength measurement).
Mechanical competence of bone can be determined using methods known
to those skilled in the art, for example, a blunt indentation
force, a three point bending to failure test or a torsional
analysis on bone samples from appropriate test subject, e.g.,
mouse, rat. Percent (%) change in bone strength can be calculated
using the following formula:
% change=[(BS.sub.2-BS.sub.1)/BS.sub.1].times.100
where BS.sub.1 is the first bone strength measurement, and BS.sub.2
is the second bone strength measurement. An increase in bone
strength is identified where the change in bone strength is greater
than 0, i.e., a positive % change. In some embodiments, increased
bone strength affected by treatment with the compound is an
increase in bone strength of at least about 1%. In other
embodiments, requisite increased bone strength affected by
treatment with the compound is an increase in bone strength of at
least about 5%, at least about 10%, at least about 15%, at least
about 20%, at least about 25%, at least about 30%, at least about
40%, at least about 50%, at least about 75%, at least about 100%,
or at least about 200%, when a first bone strength measurement and
a second bone strength measurement are compared.
[0117] In some embodiments, for example, where bone strength is
being assessed in a human subject, fracture incidence can be
recorded, and increased bone strength can be identified where there
is a trend of decreased incidence of bone fracture.
[0118] Although not necessary to establish anabolic effect,
additional information to establish promotion of net bone formation
can be obtained. For example, assays can be conducted for certain
biomarkers of bone formation (See, e.g., M J Seibel Clin Biochem
Rev 26:97 (2005) or "The Use of Biochemical Markers of Bone
Turnover in Osteoporosis" by PD Delmas et al. Osteoporosis Int.
suppl. 6S2-17 (2000), which are incorporated herein by these
references). As another example, information can be collected as
described in Riggs B L, and Parfitt A M, "Drugs Used to Treat
Osteoporosis: The Critical Need for a Uniform Nomenclature Based on
Their Action on Bone Remodeling," J. Bone and Mineral Res. 20:2
(2005), which is incorporated herein by this reference. In this
regard, in some embodiments, anabolic effect can be identified
where a biomarker of bone formation is found in an appropriate test
sample, e.g., osteocalcin, collagen type I, as described in the
Examples herein. In some embodiments, anabolic effect can be
identified pursuant to an assay to evaluate stimulation of bone
formation, e.g., calvarial injection, as described in the Examples
herein.
[0119] As used herein, an antimicrobial effect is an effect
resulting in a treatment (as defined herein) of a microbial
infection, including a bacterial infection. In some embodiments,
the compound can interact with and affect bone by treating a
microbial infection associated with bone. In some embodiments, an
anti-microbial effect includes preventing infection by a
microorganism, or inhibiting the growth of a microorganism, such as
a bacteria, or by exerting a direct killing effect on a
microorganism.
[0120] As used herein, an anti-cancer effect is an effect resulting
in a treatment (as defined herein) of a cancer. In some
embodiments, an anti-cancer effect includes slowing the growth of
or killing cancerous cells. In some embodiments, an anti-cancer
effect includes preventing the metastasis of a primary cancer to
bone. In some embodiments, an anti-cancer effect includes enhancing
the killing of cancerous cells associated with the bone (e.g.,
acting as a chemo sensitizer or radiosensitizer). With regard to
cancer treatment, compounds including an anti-cancer agent can be
effective against primary bone cancer, (e.g., primary bone sarcoma)
and/or against secondary bone cancer, i.e., metastatic bone cancer.
In the case of secondary bone cancer, breast, lung, prostate,
kidney, and thyroid cancers are the types of primary cancers that
most commonly metastasize to bone. It is contemplated that an
anti-cancer agent from which a Bone Active Portion of a compound of
the presently-disclosed subject matter is derived can be selected
based on the primary tumor site. For example, when compounds of the
presently-disclosed subject matter are used for treating secondary
cancer, the efficacy can be enhanced by selecting a Bone Active
Portion that is particularly effective against the primary cancer
type that has metastasized, or has the potential to metastasize, to
bone.
[0121] The foregoing paragraphs include information about certain
conditions or interest and/or desired effects; however, other
conditions of interest and/or desired effects are contemplated by
the presently-disclosed subject matter. For example, in some
embodiments, compounds of the presently-disclosed subject matter
can be useful for treating muscle atrophy, or loss of muscle mass,
of muscles surrounding bones of the axial skeleton. In some
embodiments, compounds wherein the Bone Active Portion is derived
from a steroid, such as testosterone, can be useful for treating
muscle atrophy (e.g., the compound of Formula 33, set forth below,
which includes a Bone Active Portion derived from testosterone, can
be useful for treating muscle atrophy). For another example, in
some embodiments, compounds of the presently-disclosed subject
matter can be useful to facilitate delivery of a second compound of
interest. For example, in some embodiments, the second compound can
be administered substantially concurrently with a compound of the
presently-disclosed subject matter that include a Bone Active
Portion that is a vasodilator, or a Bone Active Portion that is
derived from a Bone Active Agent that is a vasodilator. Without
wishing to be bound by theory or mechanism, it is believed that an
increased delivery of blood to bone affected by a bone-targeted
vasodilator of the presently-disclosed subject matter can
facilitate delivery to bone of a second compound of interest. In
some embodiments, a bone-targeted vasodilator of the
presently-disclosed subject matter includes a Bone Active Portion
that is derived from a nitric oxide agent (NO donor), such as a
nitric oxide agent as set forth in Table C.
Protecting Group (R.sub.A)
[0122] With reference to Formula 1, in some embodiments of the
compound, R.sub.A is a protecting group. The protecting group can
assist with maintaining the stability of the compound, for example,
by keeping an adjacent group on the linking portion from reacting
with other portions of the compound, e.g., cyclizing. Compounds
including a protecting group can be stably stored until it becomes
desirable to associate the compound with a Bone Active Portion. In
this regard, the compounds including a protecting group are useful
for preparing compounds for treating conditions associated with
bone.
[0123] Exemplary protecting groups that can be used include,
t-butoxycarbonyl (t-BOC) or t-butoxy, fluorenylmethoxycarbonyl
(FMOC) or a fluorenylmethoxy, and other appropriate protecting
groups, such as those described in Greene's Protective Groups in
Organic Synthesis, 4th ed., by Peter G. M. Wuts, and Theodora W.
Greene, John Wiley & Sons, inc., Hoboken, N.J., 2006, which is
incorporated herein by this reference. As will be understood by
those of ordinary skill in the art, appropriate protecting groups
can be selected for use with the compounds disclosed herein, which
will allow the compound to be stably stored, and which will allow
the compound to be used to prepare compounds for treating
conditions associated with bone, i.e., allow for the protecting
group to be removed and for a bone targeting portion to be
associated with the compound. In this regard, as will be understood
by those of ordinary skill in the art, while t-butoxycarbonyl
(t-BOC) or t-butoxy, or fluorenylmethoxycarbonyl (FMOC) or a
fluorenylmethoxy can be appropriate protecting groups in some
embodiments, they are not appropriate in other embodiments.
[0124] In some embodiments of the presently-disclosed subject
matter where the protecting group is t-BOC, the compound can be
represented by the following formula:
##STR00050##
[0125] In some embodiments of the presently-disclosed subject
matter where the protecting group is FMOC, the compound can be
represented by the following formula:
##STR00051##
Hydrogen, or Hydroxyl (R.sub.A)
[0126] With reference to Formula 1, in some embodiments of the
compound, R.sub.A is hydrogen or hydroxyl, depending on the
embodiment of the Linking Portion, R.sub.L, being used, as will be
described below. For example, in some embodiments, the compound can
be represented by the formula
##STR00052##
[0127] The presently-disclosed subject matter includes salts
derived from compounds where R.sub.A is hydrogen or hydroxyl, which
salts can be stably stored until it becomes desirable to associate
the compound with a Bone Active Portion. In this regard, the
compounds in which R.sub.A is Hydrogen or Hydroxyl are useful for
preparing compounds for treating conditions associated with
bone.
[0128] In embodiments wherein the compound is provided as a salt,
any appropriate salt known to those skilled in the art can be used.
Examples of salts include, but are not limited to: ammonium,
bromide, chloride, ethylenediamine, maleate, phosphate, sulfate,
tartrate, and tosylate salts.
Linking Portion
[0129] The Linking Portion (R.sub.L) of the compound connects and
separates the Bone Targeting Portion (R.sub.T) and a Bone Active
Portion, the protecting group, or the hydrogen or hydroxyl
(R.sub.A). Without wishing to be bound by theory, in embodiments
including a Bone Active Portion, it is believed that the Linking
Portion separates the Bone Targeting Portion and the Bone Active
Portion to limit steric interference of Bone Active Portion when
interacting with bone.
[0130] The Linking Portion can be described with reference to the
following formulas:
##STR00053##
where the Linking Portion extends between R.sub.T and R.sub.A.
[0131] With regard to the linking portion of Formula 11, i can be
0, 1, 2, or 3, k can be 0, 1, 2, or 3, and p can be 0, 1, 2, 3, or
4, where i, k, and p can vary independently of one another. For
example, in an exemplary embodiment, i can be 1, k can be 2, and p
can be 0, as represented by the following formulas:
##STR00054##
[0132] In another exemplary embodiment, i can be 2, k can be 2, and
p can be 2, as represented by the following formulas:
##STR00055##
[0133] In another exemplary embodiment, i can be 3, k can be 3, and
p can be 1, as represented by the following formulas:
##STR00056##
[0134] In another exemplary embodiment, i can be 0, k can be 0, and
p can be 0, as represented by the following formulas:
##STR00057##
[0135] The groups of the linking portion identified as R.sub.q can
be hydrogen or hydroxy, and can vary independently of one another.
For example, every R.sub.q group can be hydroxy, as shown in the
following formula, where i, k, and p are each 0:
##STR00058##
[0136] The identity of each R.sub.q group is independent. For
example, one R.sub.q group can be hydrogen, while the other R.sub.q
group can hydroxy, as shown in the following formula, where i, k,
and p are each 0:
##STR00059##
[0137] The group of the linking portion identified as X can be O,
NH, S, or a covalent bond; and the group identified as X' can
likewise be O, NH, S, or a covalent bond; where X and X' can vary
independently of one another. In some embodiments, at least one of
X' and X is a covalent bond. For example, when X is O, X' is a
covalent bond, one R.sub.q group is hydrogen, the other R.sub.q
group is hydroxy, and i, k, and p are each 0, then an exemplary
compound can be represented by the following formula:
##STR00060##
[0138] For another example, when X is a covalent bond, X' is NH,
both R.sub.q groups are hydroxy, i is 2, k is 2, and p is 0, then
an exemplary compound can be represented by the following
formula:
##STR00061##
[0139] The Linking Portion can be connected to the Bone Targeting
Portion (R.sub.T) at R.sub.1, R.sub.2, or R.sub.4. For example,
when the Linking Portion is connected to the Bone Targeting Portion
(R.sub.T) at R.sub.1, the compound can be represented by the
following formula:
##STR00062##
[0140] As mentioned above, a third unit R.sub.A of the compound can
be selected from: a Bone Targeting Portion (See e.g., Tables A-D);
a protecting group; or a hydrogen. When R.sub.A is a Bone Active
Portion, it is contemplated that the Linker Portion can be bound to
the Bone Active Portion to minimize the susceptibility to
hydrolysis, e.g., ether linkage, to increase the bioavailability of
the compound. That is to say, without wishing to be bound by theory
or mechanism, if susceptibility to hydrolysis is minimized, the
compound can be delivered to and affect bone.
[0141] In an exemplary embodiment, R.sub.A can be a Bone Active
Portion derived from estradiol, as represented by the following
formula:
##STR00063##
[0142] As shown in Formula 23, the Bone Active Portion derived from
estradiol is estradiol less a hydrogen, allowing the Bone Active
Portion to be connected to the remainder of the compound, while
maintaining one or more activities generally associated with free
estradiol. In some embodiments, when the Bone Active Portion of the
compound is derived from estradiol, it is derived from the
17-.beta.-enantiomer of estradiol. Without wishing to be bound by
theory or mechanism, it is believed that the 17-.beta.-enantiomer
of estradiol is the active isomer.
[0143] Another exemplary compound can be represented by the
following formula:
##STR00064##
where the Linking Portion is connected to the Bone Targeting
Portion (R.sub.T) at R.sub.1; where R.sub.2, R.sub.3, R.sub.5, and
R.sub.6 are each H, R.sub.4 is CH.sub.3, and R.sub.7 is NH.sub.2;
where R.sub.q are each OH, i is 2, k is 2, p is 0, and X and X' are
each a covalent bond; and where R.sub.A is derived from
estradiol
[0144] Another exemplary compound can be represented by the
following formula:
##STR00065##
where the Linking Portion is connected to the Bone Targeting
Portion (R.sub.T) at R.sub.1; where R.sub.2, R.sub.3, R.sub.5, and
R.sub.6 are each H, R.sub.4 is CH.sub.3, and R.sub.7 is NH.sub.2;
where R.sub.q are each OH, i is 2, k is 2, p is 2, X' is a covalent
bond, and X is NH; and where R.sub.A is derived from Estradiol.
[0145] In some embodiments, R.sub.A can be derived from a
non-steroidal estrogenic agent. In some embodiments, R.sub.A can be
a Bone Active Portion derived from the non-steroidal estrogenic
agent, genistein, as represented by the following formula:
##STR00066##
The Bone Active Portion derived from genistein is genistein less a
hydrogen, allowing the Bone Active Portion to be connected to the
remainder of the compound, while maintaining one or more activities
generally associated with free genistein.
[0146] Another exemplary compound can be represented by the
following formula:
##STR00067##
where the Linking Portion is connected to the Bone Targeting
Portion (R.sub.T) at R.sub.1; where R.sub.2, R.sub.3, R.sub.5, and
R.sub.6 are each H, R.sub.4 is CH.sub.3, and R.sub.7 is NH.sub.2;
where R.sub.q are each OH, i is 2, k is 2, p is 0, and X' and X are
each a covalent bond; and where R.sub.A is derived from
genistein.
[0147] In some embodiments, R.sub.A can be derived from a nitric
oxide agent. In some embodiments, R.sub.A can be a Bone Active
Portion derived from the nitric-oxide-targeted/generating agent,
alkoxy-NO.sub.2, as represented by the following formula:
##STR00068##
[0148] The Bone Active Portion derived from alkoxy-NO.sub.2 is
alkoxy-NO.sub.2 less a hydrogen, allowing the Bone Active Portion
to be connected to the remainder of the compound, while maintaining
one or more activities generally associated with free
alkoxy-NO.sub.2.
[0149] Another exemplary compound can be represented by the
following formula:
##STR00069##
where the Linking Portion is connected to the Bone Targeting
Portion (R.sub.T) at R.sub.1; where R.sub.2, R.sub.3, R.sub.5, and
R.sub.6 are each H, R.sub.4 is CH.sub.3, and R.sub.7 is NH.sub.2;
where R.sub.q are each OH, i is 2, k is 2, p is 0, and X' and X are
each a covalent bond; and where R.sub.A is derived from
alkoxy-NO.sub.2.
[0150] In some embodiments, R.sub.A can be derived from an
androgen. In some embodiments, R.sub.A can be a Bone Active Portion
derived from the androgen, dehydroepiandrosterone (DHEA), as
represented by the following formula:
##STR00070##
The Bone Active Portion derived from DHEA is DHEA singly bonded to
oxygen at carbon 17, allowing the Bone Active Portion to be
connected to the remainder of the compound, while maintaining one
or more activities generally associated with free DHEA.
[0151] Another exemplary compound can be represented by the
following formula:
##STR00071##
where the Linking Portion is connected to the Bone Targeting
Portion (R.sub.T) at R.sub.1; where R.sub.2, R.sub.3, R.sub.5, and
R.sub.6 are each H, R.sub.4 is CH.sub.3, and R.sub.7 is NH.sub.2;
where R.sub.q are each OH, i is 2, k is 2, p is 2, X and X' are
each a covalent bond; and where R.sub.A is derived from DHEA.
[0152] In some embodiments, R.sub.A can be a Bone Active Portion
derived from the androgen, testosterone, as represented by the
following formula:
##STR00072##
The Bone Active Portion derived from testosterone is testosterone
less a hydrogen, allowing the Bone Active Portion to be connected
to the remainder of the compound, while maintaining one or more
activities generally associated with free testosterone.
[0153] Another exemplary compound can be represented by the
following formula:
##STR00073##
where the Linking Portion is connected to the Bone Targeting
Portion (R.sub.T) at R.sub.1; where R.sub.2, R.sub.3, R.sub.5, and
R.sub.6 are each H, R.sub.4 is CH.sub.3, and R.sub.7 is NH.sub.2;
where R.sub.q are each H, i is 0, k is 0, p is 0, and X' and X are
each a covalent bond; and where R.sub.A is derived from
testosterone.
[0154] In some embodiments, R.sub.A can be derived from a carbonic
anhydrase inhibitor. In some embodiments, R.sub.A can be a Bone
Active Portion derived from the carbonic anhydrase inhibitor,
2-amino-1,3,4-thiadiazole-5-sulfonamide, as represented by the
following formula:
##STR00074##
The Bone Active Portion derived from
2-amino-1,3,4-thiadiazole-5-sulfonamide less a hydrogen, allowing
the Bone Active Portion to be connected to the remainder of the
compound, while maintaining one or more activities generally
associated with free sulfonamide.
[0155] Another exemplary compound can be represented by the
following formula:
##STR00075##
where the Linking Portion is connected to the Bone Targeting
Portion (R.sub.T) at R.sub.1; where R.sub.2, R.sub.3, R.sub.5, and
R.sub.6 are each H, R.sub.4 is CH.sub.3, and R.sub.7 is NH.sub.2;
where R.sub.q are each H, i is 0, k is 0, p is 0, and X' and X are
each a covalent bond; and where R.sub.A is derived from
2-aminothiadiazole-5-sulfonamide.
[0156] In some embodiments, R.sub.A can be derived from an
anti-cancer agent or an antineoplastic agent. In some embodiments,
R.sub.A can be a Bone Active Portion derived from doxorubicin, as
represented by the following formula:
##STR00076##
The Bone Active Portion derived from doxorubicin is doxorubicin
less a hydrogen, allowing the Bone Active Portion to be connected
to the remainder of the compound, while maintaining one or more
activities generally associated with free doxorubicin.
[0157] Another exemplary compound can be represented by the
following formula:
##STR00077##
where the Linking Portion is connected to the Bone Targeting
Portion (R.sub.T) at R.sub.1; where R.sub.2, R.sub.3, R.sub.5, and
R.sub.6 are each H, R.sub.4 is CH.sub.3, and R.sub.7 is NH.sub.2;
where R.sub.q are each OH, i is 2, k is 2, p is 0, and X' and X are
each a covalent bond; and where R.sub.A is derived from
doxorubicin.
[0158] In some embodiments, R.sub.A can be a Bone Active Portion
derived from dasatinib, as represented by the following
formula:
##STR00078##
The Bone Active Portion derived from dasatinib is dasatinib less a
hydrogen, allowing the Bone Active Portion to be connected to the
remainder of the compound, while maintaining one or more activities
generally associated with free dasatinib.
[0159] Another exemplary compound can be represented by the
following formula:
##STR00079##
where the Linking Portion is connected to the Bone Targeting
Portion (R.sub.T) at R.sub.1; where R.sub.2, R.sub.3, R.sub.5, and
R.sub.6 are each H, R.sub.4 is CH.sub.3, and R.sub.7 is NH.sub.2;
where R.sub.q are each OH, i is 2, k is 2, p is 0, and X' and X are
each a covalent bond; and where R.sub.A is derived from
dasatinib.
[0160] In some embodiments, R.sub.A can be derived from an
antimicrobial agent. In some embodiments, R.sub.A can be a Bone
Active Portion derived from the antimicrobial agent, vancomycin, as
represented by the following formula:
##STR00080##
The Bone Active Portion derived from vancomycin is vancomycin less
a hydroxyl, allowing the Bone Active Portion to be connected to the
remainder of the compound, while maintaining one or more activities
generally associated with free vancomycin.
[0161] It is noted that when some agents are joined to the compound
as a Bone Active Portion, they donate an oxygen atom to an amide or
ester bond of the carbonyl group between X' and X of the linking
portion. Vancomycin is such an agent. When Bone Active Portions are
derived from such agents, as will be understood by those of
ordinary skill in the art, X' can be O, NH, or S; X is a covalent
bond; and p is 0.
[0162] In this regard, an exemplary compound can be represented by
the following formula:
##STR00081##
where the Linking Portion is connected to the Bone Targeting
Portion (R.sub.T) at R.sub.1; where R.sub.2, R.sub.3, R.sub.5, and
R.sub.6 are each H, R.sub.4 is CH.sub.3, and R.sub.7 is NH.sub.2;
where R.sub.q are each OH, i is 2, k is 2, p is 0, X' is NH, and X
is a covalent bond; and where R.sub.A is derived from
vancomycin.
[0163] Another exemplary compound of the presently-disclosed
subject matter can be represented by the following formula:
##STR00082##
where R.sub.A is a protecting group, which selected protecting
group is fluorenylmethoxycarbonyl (FMOC); where the Linking Portion
is connected to the Bone Targeting Portion (R.sub.T) at R.sub.1;
where R.sub.2, R.sub.3, R.sub.5, and R.sub.6 are each H, R.sub.4 is
CH.sub.3, and R.sub.7 is NH.sub.2; and where R.sub.q are each OH, i
is 2, k is 2, p is 0, X' is NH, and X is a covalent bond.
[0164] Another exemplary compound of the presently-disclosed
subject matter can be represented by the following formula:
##STR00083##
where R.sub.A is a protecting group, which selected protecting
group is t-butoxycarbonyl (t-BOC); where the Linking Portion is
connected to the Bone Targeting Portion (R.sub.T) at R.sub.1; where
R.sub.2, R.sub.3, R.sub.5, and R.sub.6 are each H, R.sub.4 is
CH.sub.3, and R.sub.7 is NH.sub.2; and where R.sub.q are each OH, i
is 2, k is 2, p is 0, X' is NH, and X is a covalent bond.
[0165] As noted above, when some agents are joined to the compound
as an R.sub.A group, they donate an oxygen atom to an amide or
ester bond of the carbonyl group between X' and X of the linking
portion. The protecting groups FMOC and t-BOC are such agents. When
R.sub.A is derived from such an agent, as will be understood by
those of ordinary skill in the art, X' can be O, NH, or S; X is a
covalent bond; and p is 0. As such, it is noted that, in Formulas
40 and 41, X' is NH, X is a covalent bond, and p is 0.
[0166] Another exemplary compound of the presently-disclosed
subject matter can be represented by the following formula:
##STR00084##
where R.sub.A is OH; where the Linking Portion is connected to the
Bone Targeting Portion (R.sub.T) at R.sub.1; where R.sub.2,
R.sub.3, R.sub.5, and R.sub.6 are each H, R.sub.4 is CH.sub.3, and
R.sub.7 is NH.sub.2; and where R.sub.q are each OH, i is 2, k is 2,
p is 0, and X' and X are each a covalent bond.
[0167] In some exemplary embodiments, the Linking Portion can be
connected to the Bone Targeting Portion (R.sub.T) at R.sub.4, as
shown in the following formula:
##STR00085##
[0168] As mentioned above, a third unit R.sub.A of the compound can
be selected from: a Bone Targeting Portion (See e.g., Tables A-D);
a protecting group; or a hydrogen. When R.sub.A is a Bone Active
Portion, it is contemplated that the Linker Portion can be bound to
the Bone Active Portion to minimize the susceptibility to
hydrolysis, e.g., ether linkage, to increase the bioavailability of
the compound. That is to say, if susceptibility to hydrolysis is
minimized, without wishing to be bound by theory or mechanism, the
compound can be delivered to and affect bone, before the Bone
Active Portion can be cleaved from the compound.
[0169] In an exemplary embodiment, R.sub.A can be a Bone Active
Portion derived from estradiol, as represented by the following
formula:
##STR00086##
[0170] The Bone Active Portion derived from estradiol is estradiol
less a hydrogen, allowing the Bone Active Portion to be connected
to the remainder of the compound, while maintaining one or more
activities generally associated with free estradiol. In some
embodiments, when the Bone Active Portion of the compound is
derived from estradiol, it is derived from the 17-.beta.-enantiomer
of estradiol. Without wishing to be bound by theory or mechanism,
it is believed that the 17-.beta.-enantiomer of estradiol is the
active isomer.
[0171] Another exemplary compound can be represented by the
following formula:
##STR00087##
where the Linking Portion is connected to the Bone Targeting
Portion (R.sub.T) at R.sub.4; where R.sub.1, R.sub.2, R.sub.3,
R.sub.5, and R.sub.6 are each H, and R.sub.7 is NH.sub.2; where
R.sub.q are each OH, i is 2, k is 2, p is 0, and X' and X are each
a covalent bond; and where R.sub.A is derived from estradiol
[0172] Another exemplary compound can be represented by the
following formula:
##STR00088##
where the Linking Portion is connected to the Bone Targeting
Portion (R.sub.T) at R.sub.4; where R.sub.1, R.sub.2, R.sub.3,
R.sub.5, and R.sub.6 are each H, and R.sub.7 is NH.sub.2; where
R.sub.q are each OH, i is 2, k is 2, p is 2, X' is a covalent bond,
and X is NH; and where R.sub.A is derived from Estradiol.
[0173] In some embodiments, R.sub.A can be a Bone Active Portion
derived from a non-steroidal estrogenic agent. In some embodiments,
R.sub.A can be a Bone Active Portion derived from the non-steroidal
estrogenic agent, genistein, as represented by the following
formula:
##STR00089##
The Bone Active Portion derived from genistein is genistein less a
hydrogen, allowing the Bone Active Portion to be connected to the
remainder of the compound, while maintaining one or more activities
generally associated with free genistein.
[0174] Another exemplary compound can be represented by the
following formula:
##STR00090##
where the Linking Portion is connected to the Bone Targeting
Portion (R.sub.T) at R.sub.4; where R.sub.1, R.sub.2, R.sub.3,
R.sub.5, and R.sub.6 are each H, and R.sub.7 is NH.sub.2; where
R.sub.q are each OH, i is 2, k is 2, p is 0, and X' and X are each
a covalent bond; and where R.sub.A is derived from genistein.
[0175] In some embodiments, R.sub.A can be a Bone Active Portion
derived from a nitric oxide agent. In some embodiments, R.sub.A can
be a Bone Active Portion derived from the nitric oxide agent,
alkoxy-NO.sub.2, as represented by the following formula:
##STR00091##
The Bone Active Portion derived from alkoxy-NO.sub.2 is
alkoxy-NO.sub.2 less a hydrogen, allowing the Bone Active Portion
to be connected to the remainder of the compound, while maintaining
one or more activities generally associated with free
alkoxy-NO.sub.2.
[0176] Another exemplary compound can be represented by the
following formula:
##STR00092##
where the Linking Portion is connected to the Bone Targeting
Portion (R.sub.T) at R.sub.4; where R.sub.1, R.sub.2, R.sub.3,
R.sub.5, and R.sub.6 are each H, and R.sub.7 is NH.sub.2; where
R.sub.q are each OH, i is 2, k is 2, p is 0, and X' and X are each
a covalent bond; and where R.sub.A is derived from
alkoxy-NO.sub.2.
[0177] In some embodiments, R.sub.A can be a Bone Active Portion
derived from an androgen. In some embodiments, R.sub.A can be a
Bone Active Portion derived from the androgen, DHEA, as represented
by the following formula:
##STR00093##
The Bone Active Portion derived from DHEA is DHEA singly bonded to
oxygen at carbon 17, allowing the Bone Active Portion to be
connected to the remainder of the compound, while maintaining one
or more activities generally associated with free DHEA.
[0178] Another exemplary compound can be represented by the
following formula:
##STR00094##
where the Linking Portion is connected to the Bone Targeting
Portion (R.sub.T) at R.sub.4; where R.sub.1, R.sub.2, R.sub.3,
R.sub.5, and R.sub.6 are each H, and R.sub.7 is NH.sub.2; where
R.sub.q are each OH, i is 2, k is 2, p is 2, and X' and X are each
a covalent bond; and where R.sub.A is derived from DHEA.
[0179] In some embodiments, R.sub.A can be a Bone Active Portion
derived from the androgen, testosterone, as represented by the
following formula:
##STR00095##
The Bone Active Portion derived from testosterone is testosterone
less a hydrogen, allowing the Bone Active Portion to be connected
to the remainder of the compound, while maintaining one or more
activities generally associated with free testosterone.
[0180] Another exemplary compound can be represented by the
following formula:
##STR00096##
where the Linking Portion is connected to the Bone Targeting
Portion (R.sub.T) at R.sub.4; where R.sub.1, R.sub.2, R.sub.3,
R.sub.5, and R.sub.6 are each H, and R.sub.7 is NH.sub.2; where
R.sub.q are each H, i is 0, k is 0, p is 0, and X' and X are each a
covalent bond; and where R.sub.A is derived from testosterone.
[0181] In some embodiments, R.sub.A can be a Bone Active Portion
derived from a carbonic anhydrase inhibitor. In some embodiments,
R.sub.A can be a Bone Active Portion derived from the carbonic
anhydrase inhibitor, 2-aminothiadiazole-5-sulfonamide, as
represented by the following formula:
##STR00097##
The Bone Active Portion derived from
2-amino-1,3,4-thiadiazole-5-sulfonamide less a hydrogen, allowing
the Bone Active Portion to be connected to the remainder of the
compound, while maintaining one or more activities generally
associated with free sulfonamide.
[0182] Another exemplary compound can be represented by the
following formula:
##STR00098##
where the Linking Portion is connected to the Bone Targeting
Portion (R.sub.T) at R.sub.4; where R.sub.1, R.sub.2, R.sub.3,
R.sub.5, and R.sub.6 are each H, and R.sub.7 is NH.sub.2; where
R.sub.q are each H, i is 0, k is 0, p is 0, and X' and X are each a
covalent bond; and where R.sub.A is derived from
2-aminothiadiazole-5-sulfonamide.
[0183] In some embodiments, R.sub.A can be a Bone Active Portion
derived from an anti-cancer agent or an antineoplastic agent. In
some embodiments, R.sub.A can be a Bone Active Portion derived from
doxorubicin, as represented by the following formula:
##STR00099##
The Bone Active Portion derived from doxorubicin is doxorubicin
less a hydrogen, allowing the Bone Active Portion to be connected
to the remainder of the compound, while maintaining one or more
activities generally associated with free doxorubicin.
[0184] Another exemplary compound can be represented by the
following formula:
##STR00100##
where the Linking Portion is connected to the Bone Targeting
Portion (R.sub.T) at R.sub.4; where R.sub.1, R.sub.2, R.sub.3,
R.sub.5, and R.sub.6 are each H, and R.sub.7 is NH.sub.2; where
R.sub.q are each OH, i is 2, k is 2, p is 0, and X' and X are each
a covalent bond; and where R.sub.A is derived from doxorubicin.
[0185] In some embodiments, R.sub.A can be a Bone Active Portion
derived from dasatinib, as represented by the following
formula:
##STR00101##
The Bone Active Portion derived from dasatinib is dasatinib less a
hydrogen, allowing the Bone Active Portion to be connected to the
remainder of the compound, while maintaining one or more activities
generally associated with free dasatinib.
[0186] Another exemplary compound can be represented by the
following formula:
##STR00102##
[0187] where the Linking Portion is connected to the Bone Targeting
Portion (R.sub.T) at R.sub.4; where R.sub.1, R.sub.2, R.sub.3,
R.sub.5, and R.sub.6 are each H, and R.sub.7 is NH.sub.2; where
R.sub.q are each OH, i is 2, k is 2, p is 0, and X' and X are each
a covalent bond; and where R.sub.A is derived from dasatinib.
[0188] In some embodiments, R.sub.A can be a Bone Active Portion
derived from an antimicrobial agent. In some embodiments, R.sub.A
can be a Bone Active Portion derived from the antimicrobial agent,
vancomycin, as represented by the following formula:
##STR00103##
The Bone Active Portion derived from vancomycin is vancomycin less
a hydroxyl, allowing the Bone Active Portion to be connected to the
remainder of the compound, while maintaining one or more activities
generally associated with free vancomycin.
[0189] As noted herein, when some agents are joined to the compound
as a Bone Active Portion, they donate an oxygen atom to an amide or
ester bond of the carbonyl group between X' and X of the linking
portion. Vancomycin is such an agent. When Bone Active Portions are
derived from such agents, as will be understood by those of
ordinary skill in the art, X' can be O, NH, or S; X is a covalent
bond; and p is 0.
[0190] In this regard, an exemplary compound can be represented by
the following formula:
##STR00104##
where the Linking Portion is connected to the Bone Targeting
Portion (R.sub.T) at R.sub.4; where R.sub.1, R.sub.2, R.sub.3,
R.sub.5, and R.sub.6 are each H, and R.sub.7 is NH.sub.2; where
R.sub.q are each OH, i is 2, k is 2, p is 0, X' is NH, and X is a
covalent bond; and where R.sub.A is derived from vancomycin.
[0191] In some embodiments, R.sub.A is a protecting group. In some
embodiments, the compound can be represented by the following
formula:
##STR00105##
where R.sub.A is a protecting group, which selected protecting
group is fluorenylmethoxycarbonyl (FMOC); where the Linking Portion
is connected to the Bone Targeting Portion (R.sub.T) at R.sub.4;
where R.sub.1, R.sub.2, R.sub.3, R.sub.5, and R.sub.6 are each H,
and R.sub.7 is NH.sub.2; where R.sub.q are each OH, i is 2, k is 2,
p is 0, X' is NH, and X is a covalent bond.
[0192] In some embodiments, the compound can be represented by the
following formula:
##STR00106##
where R.sub.A is a protecting group, which selected protecting
group is t-butoxycarbonyl (t-BOC); where the Linking Portion is
connected to the Bone Targeting Portion (R.sub.T) at R.sub.4; where
R.sub.1, R.sub.2, R.sub.3, R.sub.5, and R.sub.6 are each H, and
R.sub.7 is NH.sub.2; where R.sub.q are each OH, i is 2, k is 2, p
is 0, X' is NH, and X is a covalent bond.
[0193] As noted above, when some agents are joined to the compound
as an R.sub.A group, they donate an oxygen atom to an amide or
ester bond of the carbonyl group between X' and X of the linking
portion. The protecting groups FMOC and t-BOC are such agents. When
R.sub.A is derived from such an agent, as will be understood by
those of ordinary skill in the art, X' can be O, NH, or S; X is a
covalent bond; and p is 0. As such, it is noted that, in Formulas
61 and 62, X' is NH, X is a covalent bond, and p is 0.
[0194] In some embodiments, R.sub.A can be a hydroxyl. In some
embodiments, the compound can be represented by the following
formula:
##STR00107##
where R.sub.A is OH; where the Linking Portion is connected to the
Bone Targeting Portion (R.sub.T) at R.sub.4; where R.sub.1,
R.sub.2, R.sub.3, R.sub.5, and R.sub.6 are each H, and R.sub.7 is
NH.sub.2; where R.sub.q are each OH, i is 2, k is 2, p is 0, and X
is a covalent bond.
[0195] Embodiments of the linking portion of Formula 12 and Formula
13 will now be described. The length of the linking portion can
vary, depending on the embodiment of the presently-disclosed
subject matter. With regard to the embodiments of the linking
portion of Formula 12 and Formula 13, m can be 1 to about 3, and n
can be 1 to about 4. For example, when m=1 and n=2, the compounds
can be represented by the following formulas:
##STR00108##
[0196] For another example, when m=1 and n=3, the compounds can be
represented by the following formulas:
##STR00109##
[0197] When m>1, multiple n-groups are provided. When multiple
n-groups are provided, each n is independently 1, 2, 3, or 4. For
example, when m=3, three n-groups are provided, n', n'', and n''',
which are each independently 1, 2, 3, or 4 4. With regard to the
linking portion of Formula 12, when m=2, two n-groups are provided,
n' and n'', as shown in the following formula:
##STR00110##
In the compound of Formula 72, n' and n'' can each independently be
1, 2, 3, or 4. For example, when m=2, n'=1, and n''=2, a compound
according to the following formula is provided:
##STR00111##
[0198] With regard to the linking portion of Formula 13, X can be
O, NH, S, or covalent bond. When m>1, there will be more than
one X; however, in such cases, only the X of the m-group adjacent
R.sub.A can be something other than a covalent bond. In the
following formula where m=2, two n-groups are provided, n' and n'',
and two X are provided, X' and X'':
##STR00112##
Each n-group, n' and n'', of Formula 74 is independently 1 to about
4. However, only the X of the m-group adjacent R.sub.A can be
something other than a covalent bond. As such, X' must be a
covalent bond, but X'', which is the X of the m-group adjacent
R.sub.A, can be O, NH, S, or covalent bond. For example, in the
compound of the following formula, m=2, n'=1, and n''=2, X' is a
covalent bond, and X'' is NH:
##STR00113##
[0199] The groups of the linking portion identified as R.sub.S can
be hydrogen, hydroxy, or lower alkyl. For example, every R.sub.S
group could be hydrogen, as shown in the following formulas, where
m is 1, and n is 2:
##STR00114##
[0200] The identity of each R.sub.S group is independent. For
example, certain of the R.sub.S groups could be hydrogen, while
others could be hydroxy, as shown in the following formula, where m
is 1 and n is 2; and where X is a covalent bond for Formula 79:
##STR00115##
[0201] Other exemplary compounds of the presently-disclosed subject
matter can be represented by the following formulas:
##STR00116##
where certain of the R.sub.S groups are hydrogen, other R.sub.S
groups are hydroxy, another of the R.sub.S groups is methyl, m is
1, and n is 2; and where X is a covalent bond for Formula 81.
[0202] The groups of the linking portion identified as D, E, and G
are as follows.
[0203] D and G are independently selected from: covalent bond;
##STR00117##
other functional groups capable of reacting with an amine, less a
leaving group. That is to say, for example, an acyl halide
(Y--C.dbd.O) is a functional group capable of reacting with an
amine, and a carbonyl
##STR00118##
is an acyl halide, less a halogen atom (Y).
[0204] E is selected from: covalent bond; --(CT.sub.2).sub.n--,
where T is H, OH, or lower alkyl, and r=1 to about 8; and
--(C).sub.r--, where r=2 to about 8, and where the carbons are
unsaturated or partially saturated with H.
[0205] In some embodiments of the linking portion of Formula 13 and
Formula 14, when D, E, and G are all covalent bond, X is NH or O,
and R.sub.S is H; or when D is carbonyl, E and G are covalent bond,
and R.sub.S is H; then n is 2, 3, or 4.
[0206] Exemplary compounds of the presently-disclosed subject
matter can be represented by the following formulas:
##STR00119##
where D is
##STR00120##
E is --(CH.sub.2).sub.2--, and G is a covalent bond.
[0207] D, E, and G can be selected, for example, based on the
portion of the compound to which the linking group will be bound.
For example, when the Linking Portion of Formula 13 is used, the
-D-E-G- segment of the Linking Portion is adjacent the Bone
Targeting Portion of the compound. The Linking Portion can be
connected to the Bone Targeting Portion (R.sub.T) at R.sub.1, as
shown in the following formula:
##STR00121##
[0208] When the Linking Portion of Formula 13 is connected to the
Bone Targeting Portion (R.sub.T) at R.sub.1, as will be understood
by those skilled in the art, it can be beneficial for G to be
selected to be a functional group capable of reacting with an
amine, less a leaving group, for example,
##STR00122##
In this regard, an exemplary compound of the presently-disclosed
subject matter can be represented by the following formula:
##STR00123##
where the Linking Portion of Formula 13 is connected to the Bone
Targeting Portion (R.sub.T) at R.sub.1; where D and G are each
##STR00124##
and where E is --(CH.sub.2).sub.2--.
[0209] As mentioned above, a third unit R.sub.A of the compound can
be selected from: a Bone Targeting Portion (See e.g., Tables A-D);
a protecting group; or a hydroxyl, when Linking Portion of Formula
13 is selected.
[0210] In some embodiments, R.sub.A can be a Bone Active Portion
derived from a steroidal estrogenic agent. In some embodiments,
R.sub.A can be a Bone Active Portion derived from the estradiol, as
represented by the following formula, where D and G are each
##STR00125##
E is --(CH.sub.2).sub.2--, and X is a covalent bond:
##STR00126##
[0211] The Bone Active Portion derived from estradiol is estradiol
less a hydrogen, allowing the Bone Active Portion to be connected
to the remainder of the compound, while maintaining one or more
activities generally associated with free estradiol. In some
embodiments, when the Bone Active Portion of the compound is
derived from estradiol, it is derived from the 17-.beta.-enantiomer
of estradiol. Without wishing to be bound by theory or mechanism,
it is believed that the 17-.beta.-enantiomer of estradiol is the
active isomer.
[0212] Another exemplary compound of the presently-disclosed
subject matter can be represented by the following formula:
##STR00127##
where the Linking Portion is connected to the Bone Targeting
Portion (R.sub.T) at R.sub.1; D and G are each
##STR00128##
E is --(CH.sub.2).sub.2--; m is 1; n is 2; X is a covalent bond;
each R.sub.S is hydrogen; R.sub.2, R.sub.3, R.sub.5, R.sub.6 are
each H; R.sub.4 is methyl; R.sub.7 is amino; and where R.sub.A is
derived from estradiol.
[0213] In some embodiments, R.sub.A can be a Bone Active Portion
derived from a non-steroidal estrogenic agent. In some embodiments,
R.sub.A can be a Bone Active Portion derived from the non-steroidal
estrogenic agent, genistein, as represented by the following
formula, where D and G are each
##STR00129##
E is --(CH.sub.2).sub.2--, and X is a covalent bond:
##STR00130##
The Bone Active Portion derived from genistein is genistein less a
hydrogen, allowing the Bone Active Portion to be connected to the
remainder of the compound, while maintaining one or more activities
generally associated with free genistein.
[0214] Another exemplary compound of the presently-disclosed
subject matter can be represented by the following formula:
##STR00131##
where the Linking Portion is connected to the Bone Targeting
Portion (R.sub.T) at R.sub.1; D and G are each
##STR00132##
E is --(CH.sub.2).sub.2--; m is 1; n is 2; X is a covalent bond;
each R.sub.S is hydrogen; R.sub.2, R.sub.3, R.sub.5, R.sub.6 are
each H; R.sub.4 is methyl; R.sub.7 is amino; and where R.sub.A is
derived from genistein.
[0215] In some embodiments, R.sub.A can be a Bone Active Portion
derived from a nitric oxide agent. In some embodiments, R.sub.A can
be a Bone Active Portion derived from the nitric oxide agent,
alkoxy-NO.sub.2, as represented by the following formula, where D
and G are each
##STR00133##
E is --(CH.sub.2).sub.2--, and X is a covalent bond:
[0216] :
##STR00134##
The Bone Active Portion derived from alkoxy-NO.sub.2 is
alkoxy-NO.sub.2 less a hydrogen, allowing the Bone Active Portion
to be connected to the remainder of the compound, while maintaining
one or more activities generally associated with free
alkoxy-NO.sub.2.
[0217] Another exemplary compound of the presently-disclosed
subject matter can be represented by the following formula:
##STR00135##
where the Linking Portion is connected to the Bone Targeting
Portion (R.sub.T) at R.sub.1; D and G are each
##STR00136##
E is --(CH.sub.2).sub.2--; m is 1; n is 2; X is a covalent bond;
each R.sub.S is hydrogen; R.sub.2, R.sub.3, R.sub.5, R.sub.6 are
each H; R.sub.4 is methyl; R.sub.7 is amino; and where R.sub.A is
derived from alkoxy-NO.sub.2.
[0218] In some embodiments, R.sub.A can be a Bone Active Portion
derived from an androgen. In some embodiments, R.sub.A can be a
Bone Active Portion derived from the androgen, DHEA, as represented
by the following formula, where D and G are each
##STR00137##
E is --(CH.sub.2).sub.2--, and X is a covalent bond:
##STR00138##
The Bone Active Portion derived from DHEA is DHEA singly bonded to
oxygen at carbon 17, allowing the Bone Active Portion to be
connected to the remainder of the compound, while maintaining one
or more activities generally associated with free DHEA.
[0219] Another exemplary compound of the presently-disclosed
subject matter can be represented by the following formula:
##STR00139##
[0220] where the Linking Portion is connected to the Bone Targeting
Portion (R.sub.T) at R.sub.1; D and G are each
##STR00140##
E is --(CH.sub.2).sub.2--; m is 1; n is 2; X is a covalent bond;
each R.sub.S is hydrogen; R.sub.2, R.sub.3, R.sub.5, R.sub.6 are
each H; R.sub.4 is methyl; R.sub.7 is amino; and where R.sub.A is
derived from DHEA.
[0221] In some embodiments, R.sub.A can be a Bone Active Portion
derived from the androgen, testosterone, as represented by the
following formula, where D and G are each
##STR00141##
E is --(CH.sub.2).sub.2--, and X is a covalent bond:
##STR00142##
The Bone Active Portion derived from testosterone is testosterone
less a hydrogen, allowing the Bone Active Portion to be connected
to the remainder of the compound, while maintaining one or more
activities generally associated with free testosterone.
[0222] Another exemplary compound of the presently-disclosed
subject matter can be represented by the following formula:
##STR00143##
where the Linking Portion is connected to the Bone Targeting
Portion (R.sub.T) at R.sub.1; where: D and G are each
##STR00144##
E is --(CH.sub.2).sub.2--; m is 1; n is 2; X is a covalent bond;
each R.sub.S is hydrogen; R.sub.2, R.sub.3, R.sub.5, R.sub.6 are
each H; R.sub.4 is methyl; R.sub.7 is amino; and where R.sub.A is
derived from testosterone.
[0223] In some embodiments, R.sub.A can be a Bone Active Portion
derived from a carbonic anhydrase inhibitor. In some embodiments,
R.sub.A can be a Bone Active Portion derived from the carbonic
anhydrase inhibitor, 2-aminothiadiazole-5-sulfonamide, as
represented by the following formula, where D and G are each
##STR00145##
E is --(CH.sub.2).sub.2--, and X is a covalent bond:
##STR00146##
The Bone Active Portion derived from
2-amino-1,3,4-thiadiazole-5-sulfonamide is 2-amino-1,
3,4-thiadiazole-5-sulfonamide less a hydrogen, allowing the Bone
Active Portion to be connected to the remainder of the compound,
while maintaining one or more activities generally associated with
free sulfonamide.
[0224] Another exemplary compound of the presently-disclosed
subject matter can be represented by the following formula:
##STR00147##
where the Linking Portion is connected to the Bone Targeting
Portion (R.sub.T) at R.sub.1; where: D and G are each
##STR00148##
E is --(CH.sub.2).sub.2--; m is 1; n is 2; X is a covalent bond;
each R.sub.S is hydrogen; R.sub.2, R.sub.3, R.sub.5, R.sub.6 are
each H; R.sub.4 is methyl; R.sub.7 is amino; and where R.sub.A is
derived from 2-aminothiadiazole-5-sulfonamide.
[0225] In some embodiments, R.sub.A can be a Bone Active Portion
derived from an anti-cancer agent or an antineoplastic agent. In
some embodiments, R.sub.A can be a Bone Active Portion derived from
doxorubicin, as represented by the following formula, where D and G
are each
##STR00149##
E is --(CH.sub.2).sub.2--, and X is a covalent bond:
##STR00150##
The Bone Active Portion derived from doxorubicin is doxorubicin
less a hydrogen, allowing the Bone Active Portion to be connected
to the remainder of the compound, while maintaining one or more
activities generally associated with free doxorubicin.
[0226] Another exemplary compound of the presently-disclosed
subject matter can be represented by the following formula:
##STR00151##
where the Linking Portion is connected to the Bone Targeting
Portion (R.sub.T) at R.sub.1; where: D and G are each
##STR00152##
E is --(CH.sub.2).sub.2--; m is 1; n is 2; X is a covalent bond;
each R.sub.S is hydrogen; R.sub.2, R.sub.3, R.sub.5, R.sub.6 are
each H; R.sub.4 is methyl; R.sub.7 is amino; and where R.sub.A is
derived from doxorubicin.
[0227] In some embodiments, R.sub.A can be a Bone Active Portion
derived from dasatinib, as represented by the following formula,
where D and G are each
##STR00153##
E is --(CH.sub.2).sub.2--, and X is a covalent bond:
##STR00154##
The Bone Active Portion derived from dasatinib is dasatinib less a
hydrogen, allowing the Bone Active Portion to be connected to the
remainder of the compound, while maintaining one or more activities
generally associated with free dasatinib.
[0228] Another exemplary compound of the presently-disclosed
subject matter can be represented by the following formula:
##STR00155##
[0229] where the Linking Portion is connected to the Bone Targeting
Portion (R.sub.T) at R.sub.1; where: D and G are each
##STR00156##
E is --(CH.sub.2).sub.2--; m is 1; n is 2; X is a covalent bond;
each R.sub.S is hydrogen; R.sub.2, R.sub.3, R.sub.5, R.sub.6 are
each H; R.sub.4 is methyl; R.sub.7 is amino; and where R.sub.A is
derived from dasatinib.
[0230] In some embodiments, R.sub.A can be a Bone Active Portion
derived from an antimicrobial agent. In some embodiments, R.sub.A
can be a Bone Active Portion derived from the antimicrobial agent,
vancomycin, as represented by the following formula, where D and G
are each
##STR00157##
and E is --(CH.sub.2).sub.2--:
##STR00158##
[0231] The Bone Active Portion derived from vancomycin is
vancomycin less a hydroxyl, allowing the Bone Active Portion to be
connected to the remainder of the compound, while maintaining one
or more activities generally associated with free vancomycin.
[0232] It is noted that, when some agents are joined to the
compound as a Bone Active Portion, they donate an oxygen atom to an
amide or ester bond of the carbonyl group adjacent X of the linking
portion. Vancomycin is such an agent. When Bone Active Portions are
derived from such agents, as will be understood by those of
ordinary skill in the art, the X of the m-group adjacent R.sub.A is
something other than a covalent bond, i.e., O, NH, or S. As noted
above with reference to Formula 74, when m>1, there will be more
than one X; however only the X of the m-group adjacent R.sub.A can
be something other than a covalent bond, any other X must be
covalent bond.
[0233] As such, in some embodiments, the compound of the
presently-disclosed subject matter can be represented by the
following formula:
##STR00159##
where the Linking Portion is connected to the Bone Targeting
Portion (R.sub.T) at R.sub.1; where: D and G are each
##STR00160##
E is --(CH.sub.2).sub.2--; m is 1; n is 2; X is NH; each R.sub.S is
hydrogen; R.sub.2, R.sub.3, R.sub.5, R.sub.6 are each H; R.sub.4 is
methyl; R.sub.7 is amino; and where R.sub.A is derived from
vancomycin.
[0234] In some embodiments, R.sub.A is a protecting group. In some
embodiments, the compound can be represented by the following
formula:
##STR00161##
where D and G are each
##STR00162##
E is --(CH.sub.2).sub.2--, and R.sub.A is a protecting group, which
selected protecting group is fluorenylmethoxycarbonyl (FMOC). As
noted above, when some agents are joined to the compound as an
R.sub.A group, they donate an oxygen atom to an amide or ester bond
of the carbonyl group adjacent X of the linking portion. The
protecting group FMOC is such an agent. When R.sub.A is derived
from such an agent, as will be understood by those of ordinary
skill in the art, the X of the m-group adjacent R.sub.A is
something other than a covalent bond, i.e., O, NH, or S, while any
other X must be covalent bond.
[0235] In some embodiments, R.sub.A can be a hydroxyl, as
represented by the following formula, where D and G are each
##STR00163##
E is --(CH.sub.2).sub.2--, and X is a covalent bond:
##STR00164##
[0236] When the Linking Portion of Formula 13 is used, it can also
be connected to the Bone Targeting Portion (R.sub.T) at R.sub.4, as
shown in the following formula:
##STR00165##
[0237] When the Linking Portion of Formula 13 is connected to the
Bone Targeting Portion (R.sub.T) at R.sub.4, as will be understood
by those skilled in the art, the following can be beneficial: G is
a covalent bond; and D is a functional group capable of reacting
with an amine, less a leaving group. An exemplary compound of the
presently-disclosed subject matter can be represented by the
following formula, where D and E are each a covalent bond, and G
is
##STR00166##
##STR00167##
[0238] As mentioned above, the third unit R.sub.A of the compound
can be selected from: a Bone Targeting Portion (See e.g., Tables
A-D); a protecting group; or a hydroxyl, when Linking Portion of
Formula 13 is selected.
[0239] In some embodiments, R.sub.A can be a Bone Active Portion
derived from a steroidal estrogenic agent. In some embodiments,
R.sub.A can be a Bone Active Portion derived from the steroidal
estrogenic agent, estradiol, as represented by the following
formula, where D and E are each a covalent bond, G is
##STR00168##
and X is a covalent bond:
##STR00169##
[0240] The Bone Active Portion derived from estradiol is estradiol
less a hydrogen, allowing the Bone Active Portion to be connected
to the remainder of the compound, while maintaining one or more
activities generally associated with free estradiol. In some
embodiments, when the Bone Active Portion of the compound is
derived from estradiol, it is derived from the 17-.beta.-enantiomer
of estradiol. Without wishing to be bound by theory or mechanism,
it is believed that the 17-.beta.-enantiomer of estradiol is the
active isomer.
[0241] Another exemplary compound of the presently-disclosed
subject matter can be represented by the following formula:
##STR00170##
where the Linking Portion is connected to the Bone Targeting
Portion (R.sub.T) at R.sub.4; where: D and E are each a covalent
bond; G is
##STR00171##
m is 1; n is 2; X is a covalent bond; each R.sub.S is hydrogen;
R.sub.1, R.sub.2, R.sub.3, R.sub.5, and R.sub.6 are each H; R.sub.7
is NO.sub.2; and where R.sub.A is derived from estradiol.
[0242] In some embodiments, R.sub.A can be a Bone Active Portion
derived from a non-steroidal estrogenic agent. In some embodiments,
R.sub.A can be a Bone Active Portion derived from the non-steroidal
estrogenic agent, genistein, as represented by the following
formula, where D and E are each a covalent bond, G is
##STR00172##
and X is a covalent bond:
##STR00173##
The Bone Active Portion derived from genistein is genistein less a
hydrogen, allowing the Bone Active Portion to be connected to the
remainder of the compound, while maintaining one or more activities
generally associated with free genistein.
[0243] Another exemplary compound of the presently-disclosed
subject matter can be represented by the following formula:
##STR00174##
where the Linking Portion is connected to the Bone Targeting
Portion (R.sub.T) at R.sub.4; where: D and E are each a covalent
bond; G is
##STR00175##
m is 1; n is 2; X is a covalent bond; each R.sub.S is hydrogen;
R.sub.1, R.sub.2, R.sub.3, R.sub.5, and R.sub.6 are each H; R.sub.7
is NO.sub.2; and where R.sub.A is derived from genistein.
[0244] In some embodiments, R.sub.A can be a Bone Active Portion
derived from a nitric oxide agent agent. In some embodiments,
R.sub.A can be a Bone Active Portion derived from the nitric oxide
agent, alkoxy-NO.sub.2, as represented by the following formula,
where D and E are each a covalent bond, G is
##STR00176##
and X is a covalent bond:
##STR00177##
The Bone Active Portion derived from alkoxy-NO.sub.2 is
alkoxy-NO.sub.2 less a hydrogen, allowing the Bone Active Portion
to be connected to the remainder of the compound, while maintaining
one or more activities generally associated with free
alkoxy-NO.sub.2.
[0245] Another exemplary compound of the presently-disclosed
subject matter can be represented by the following formula:
##STR00178##
where the Linking Portion is connected to the Bone Targeting
Portion (R.sub.T) at R.sub.4; where: D and E are each a covalent
bond; G is
##STR00179##
m is 1; n is 2; X is a covalent bond; each R.sub.S is hydrogen;
R.sub.1, R.sub.2, R.sub.3, R.sub.5, and R.sub.6 are each H; R.sub.7
is NO.sub.2; and where R.sub.A is derived from alkoxy-NO.sub.2.
[0246] In some embodiments, R.sub.A can be a Bone Active Portion
derived from an androgen agent. In some embodiments, R.sub.A can be
a Bone Active Portion derived from the androgen, DHEA, as
represented by the following formula, where D and E are each a
covalent bond, G is
##STR00180##
and X is a covalent bond:
##STR00181##
The Bone Active Portion derived from DHEA is DHEA singly bonded to
oxygen at carbon 17, allowing the Bone Active Portion to be
connected to the remainder of the compound, while maintaining one
or more activities generally associated with free DHEA.
[0247] Another exemplary compound of the presently-disclosed
subject matter can be represented by the following formula:
##STR00182##
where the Linking Portion is connected to the Bone Targeting
Portion (R.sub.T) at R.sub.4; where: D and E are each a covalent
bond; G is
##STR00183##
m is 1; n is 2; X is a covalent bond; each R.sub.S is hydrogen;
R.sub.1, R.sub.2, R.sub.3, R.sub.5, and R.sub.6 are each H; R.sub.7
is NO.sub.2; and where R.sub.A is derived from DHEA.
[0248] In some embodiments, R.sub.A can be a Bone Active Portion
derived from the androgen, testosterone, as represented by the
following formula, where D and E are each a covalent bond, G is
##STR00184##
and X is a covalent bond:
##STR00185##
The Bone Active Portion derived from testosterone is testosterone
less a hydrogen, allowing the Bone Active Portion to be connected
to the remainder of the compound, while maintaining one or more
activities generally associated with free testosterone.
[0249] Another exemplary compound of the presently-disclosed
subject matter can be represented by the following formula:
##STR00186##
where the Linking Portion is connected to the Bone Targeting
Portion (R.sub.T) at R.sub.4; where: D and E are each a covalent
bond; G is
##STR00187##
m is 1; n is 2; X is a covalent bond; each R.sub.S is hydrogen;
R.sub.1, R.sub.2, R.sub.3, R.sub.5, and R.sub.6 are each H; R.sub.7
is NO.sub.2; and where R.sub.A is derived from testosterone.
[0250] In some embodiments, R.sub.A can be a Bone Active Portion
derived from a carbonic anhydrase inhibitor. In some embodiments,
R.sub.A can be a Bone Active Portion derived from the carbonic
anhydrase inhibitor, 2-aminothiadiazole-5-sulfonamide, as
represented by the following formula, where D and E are each a
covalent bond, G is
##STR00188##
and X is a covalent bond:
##STR00189##
The Bone Active Portion derived from
2-amino-1,3,4-thiadiazole-5-sulfonamide is 2-amino-1,
3,4-thiadiazole-5-sulfonamide less a hydrogen, allowing the Bone
Active Portion to be connected to the remainder of the compound,
while maintaining one or more activities generally associated with
free sulfonamide.
[0251] Another exemplary compound of the presently-disclosed
subject matter can be represented by the following formula:
##STR00190##
where the Linking Portion is connected to the Bone Targeting
Portion (R.sub.T) at R.sub.4; where: D and E are each a covalent
bond; G is
##STR00191##
m is 1; n is 2; X is a covalent bond; each R.sub.S is hydrogen;
R.sub.1, R.sub.2, R.sub.3, R.sub.5, and R.sub.6 are each H; R.sub.7
is NO.sub.2; and where R.sub.A is derived from
2-aminothiadiazole-5-sulfonamide.
[0252] In some embodiments, R.sub.A can be a Bone Active Portion
derived from an anti-cancer agent or an antineoplastic agent. In
some embodiments, R.sub.A can be a Bone Active Portion derived from
doxorubicin, as represented by the following formula, where D and E
are each a covalent bond, G is
##STR00192##
and X is a covalent bond:
##STR00193##
The Bone Active Portion derived from doxorubicin is doxorubicin
less a hydrogen, allowing the Bone Active Portion to be connected
to the remainder of the compound, while maintaining one or more
activities generally associated with free doxorubicin.
[0253] Another exemplary compound of the presently-disclosed
subject matter can be represented by the following formula:
##STR00194##
where the Linking Portion is connected to the Bone Targeting
Portion (R.sub.T) at R.sub.4; where: D and E are each a covalent
bond; G is
##STR00195##
m is 1; n is 2; X is a covalent bond; each R.sub.S is hydrogen;
R.sub.1, R.sub.2, R.sub.3, R.sub.5, and R.sub.6 are each H; R.sub.7
is NO.sub.2; and where R.sub.A is derived from doxorubicin.
[0254] In some embodiments, R.sub.A can be a Bone Active Portion
derived from dasatinib, as represented by the following formula,
where D and E are each a covalent bond, G is
##STR00196##
and X is a covalent bond:
##STR00197##
The Bone Active Portion derived from dasatinib is dasatinib less a
hydrogen, allowing the Bone Active Portion to be connected to the
remainder of the compound, while maintaining one or more activities
generally associated with free dasatinib.
[0255] Another exemplary compound of the presently-disclosed
subject matter can be represented by the following formula:
##STR00198##
where the Linking Portion is connected to the Bone Targeting
Portion (R.sub.T) at R.sub.4; where: D and E are each a covalent
bond; G is
##STR00199##
m is 1; n is 2; X is a covalent bond; each R.sub.S is hydrogen;
R.sub.1, R.sub.2, R.sub.3, R.sub.5, and R.sub.6 are each H; R.sub.7
is NO.sub.2; and where R.sub.A is derived from dasatinib.
[0256] In some embodiments, R.sub.A can be a Bone Active Portion
derived from an antimicrobial agent. In some embodiments, R.sub.A
can be a Bone Active Portion derived from the antimicrobial agent,
vancomycin, as represented by the following formula, where D and E
are each a covalent bond, and G is
##STR00200##
##STR00201##
The Bone Active Portion derived from vancomycin is vancomycin less
a hydroxyl, allowing the Bone Active Portion to be connected to the
remainder of the compound, while maintaining one or more activities
generally associated with free vancomycin.
[0257] As noted above, when some agents are joined to the compound
as a Bone Active Portion, they donate an oxygen atom to an amide or
ester bond of the carbonyl group adjacent X of the linking portion.
Vancomycin is such an agent. When Bone Active Portions are derived
from such agents, as will be understood by those of ordinary skill
in the art, the X of the m-group adjacent R.sub.A is something
other than a covalent bond, i.e., O, NH, or S. As noted above with
reference to Formula 74, when m>1, there will be more than one
X; however only the X of the m-group adjacent R.sub.A can be
something other than a covalent bond, any other X must be covalent
bond.
[0258] Another exemplary compound of the presently-disclosed
subject matter can be represented by the following formula:
##STR00202##
where the Linking Portion is connected to the Bone Targeting
Portion (R.sub.T) at R.sub.4; where: D and E are each a covalent
bond; G is
##STR00203##
m is 1; n is 2; X is NH; each R.sub.S is hydrogen; R.sub.1,
R.sub.2, R.sub.3, R.sub.5, and R.sub.6 are each H; R.sub.7 is
NO.sub.2; and where R.sub.A is derived from vancomycin.
[0259] In some embodiments, R.sub.A is a protecting group. In some
embodiments, the compound can be represented by the following
formula:
##STR00204##
Where D and E are each a covalent bond; G is
##STR00205##
m is 1; n is 2; X is NH; each R.sub.S is hydrogen; and R.sub.1,
R.sub.2, R.sub.3, R.sub.5, and R.sub.6 are each H; R.sub.7 is
NO.sub.2; and R.sub.A is a protecting group, which selected
protecting group is fluorenylmethoxycarbonyl (FMOC). As noted
above, when some agents are joined to the compound as an R.sub.A
group, they donate an oxygen atom to an amide or ester bond of the
carbonyl group adjacent X of the linking portion. The protecting
group FMOC is such an agent. When R.sub.A is derived from such an
agent, as will be understood by those of ordinary skill in the art,
the X of the m-group adjacent R.sub.A is something other than a
covalent bond, i.e., O, NH, or S, while any other X must be
covalent bond.
[0260] In some embodiments, R.sub.A can be a hydroxyl. In some
embodiments, the compound can be represented by the following
formula:
##STR00206##
where D and E are each a covalent bond, G is
##STR00207##
and X is a covalent bond; each R.sub.S is hydrogen; and R.sub.1,
R.sub.2, R.sub.3, R.sub.5, and R.sub.6 are each H; R.sub.7 is
NO.sub.2; and R.sub.A is hydroxyl.
[0261] With reference again to the -D-E-G- segment of the Linking
Portion, when the Linking Portion of Formula 12 is used, the
-D-E-G- segment of the Linking Portion is adjacent the R.sub.A
portion of the compounds, as shown in the following formula, where
the Linking Portion is connected to the Bone Targeting Portion at
R.sub.1:
##STR00208##
Because the -D-E-G- segment is positioned adjacent the third unit,
R.sub.A, it can be beneficial to select -D-E-G- based on the
selected third unit, R.sub.A. For example, when R.sub.A is a Bone
Active Portion, it is contemplated that the Linker Portion can be
bound to the Bone Active Portion to minimize the susceptibility to
hydrolysis, e.g., ester, urido, ether, linkage, to increase the
bioavailablity of the compound. That is to say, if susceptibility
to hydrolysis is minimized, without wishing to be bound by theory
or mechanism, the compound can be delivered to and affect bone.
[0262] As mentioned above, the third unit R.sub.A of the compound
can be selected from: a protecting group; a hydrogen when the
Linking Portion of Formula 12 (R.sub.L Option 1) is selected; or a
Bone Targeting Portion (See e.g., Tables A-D).
[0263] When the protecting group t-butoxycarbonyl (t-BOC) is used,
as will be understood by those skilled in the art, it can be
beneficial in some embodiments for D, E, and G to be a covalent
bond. In this regard, in some embodiments, the compound of the
presently-disclosed subject matter can be represented by the
following formula:
##STR00209##
where D, E, and G are covalent bonds, and R.sub.A is t-BOC.
[0264] In some embodiments, the compound of the presently-disclosed
subject matter can be represented by the following formula:
##STR00210##
where D, E, and G are a covalent bond, and R.sub.A is H.
[0265] In some embodiments, R.sub.A can be derived from a bone
active agent. In some embodiments, R.sub.A can be a Bone Active
Portion derived from a steroidal estrogenic agent. In some
embodiments, R.sub.A can be a Bone Active Portion derived from the
a steroidal estrogenic agent, estradiol. As will be understood by
those skilled in the art, in such cases, it can be beneficial in
some embodiments for D to be
##STR00211##
for E to be a group other than a covalent bond, and for G to be a
covalent bond. In this regard, in some embodiments, the compound
can be represented by the following formula, where D is
##STR00212##
E is --(CH.sub.2).sub.2--, and G is a covalent bond, and R.sub.A is
derived from estradiol:
##STR00213##
[0266] The Bone Active Portion derived from estradiol is estradiol
less a hydrogen, allowing the Bone Active Portion to be connected
to the remainder of the compound, while maintaining one or more
activities generally associated with free estradiol. In some
embodiments, when the Bone Active Portion of the compound is
derived from estradiol, it is derived from the 17-.beta.-enantiomer
of estradiol. Without wishing to be bound by theory or mechanism,
it is believed that the 17-.beta.-enantiomer of estradiol is the
active isomer.
[0267] Another exemplary compound of the presently-disclosed
subject matter can be represented by the following formula:
##STR00214##
where D is
##STR00215##
E is --(CH.sub.2).sub.2--; G is a covalent bond; m is 1; n is 2;
each R.sub.S is hydrogen; R.sub.2, R.sub.3, R.sub.5, R.sub.6 are
each H; R.sub.4 is methyl; R.sub.7 is amino; and where R.sub.A is
derived from estradiol.
[0268] Another exemplary compound of the presently-disclosed
subject matter can be represented by the following formula:
##STR00216##
where D is
##STR00217##
E is --(CH.sub.2).sub.2--; G is a covalent bond; m is 1; n is 3;
each R.sub.S is hydrogen; R.sub.2, R.sub.3, R.sub.5, R.sub.6 are
each H; R.sub.4 is methyl; R.sub.7 is amino; and where R.sub.A is
derived from estradiol.
[0269] Another exemplary compound of the presently-disclosed
subject matter can be represented by the following formula:
##STR00218##
where D is
##STR00219##
E is --(CH.sub.2).sub.2--; G is a covalent bond; m is 1; n is 2;
each R.sub.S is hydrogen; R.sub.2, R.sub.3, R.sub.5, R.sub.6 are
each H; R.sub.4 is benzyl; R.sub.7 is amino; and where R.sub.A is
derived from estradiol.
[0270] Another exemplary compound of the presently-disclosed
subject matter can be represented by the following formula:
##STR00220##
where D is
##STR00221##
E is --(CH.sub.2).sub.2--; G is a covalent bond; m is 1; n is 2;
each R.sub.S is hydrogen; R.sub.2, R.sub.3, R.sub.5, R.sub.4,
R.sub.6 are each H; R.sub.7 is amino; and where R.sub.A is derived
from estradiol.
[0271] In some embodiments, R.sub.A can be a Bone Active Portion
derived from an non-steroidal estrogenic agent. In some
embodiments, R.sub.A can be a Bone Active Portion derived from the
non-steroidal estrogenic agent, genistein, as represented by the
following formulas:
##STR00222##
##STR00223##
The Bone Active Portion derived from genistein is genistein less a
hydrogen, allowing the Bone Active Portion to be connected to the
remainder of the compound, while maintaining one or more activities
generally associated with free genistein.
[0272] Another exemplary compound of the presently-disclosed
subject matter can be represented by the following formulas:
##STR00224##
where the Linking Portion is connected to the Bone Targeting
Portion (R.sub.T) at R.sub.1; where: D is
##STR00225##
E is --(CH.sub.2)--; G is a covalent bond; m is 1; n is 2; each
R.sub.S is hydrogen; R.sub.2, R.sub.3, R.sub.5, R.sub.6 are each H;
R.sub.4 is methyl; R.sub.7 is amino; and where R.sub.A is derived
from genistein; and
##STR00226##
where the Linking Portion is connected to the Bone Targeting
Portion (R.sub.T) at R.sub.1; where: D and G are each
##STR00227##
E is --(CH.sub.2).sub.3--; m is 1; n is 2; each R.sub.S is
hydrogen; R.sub.2, R.sub.3, R.sub.5, R.sub.6 are each H; R.sub.4 is
methyl; R.sub.7 is amino; and where R.sub.A is derived from
genistein.
[0273] In some embodiments, R.sub.A can be a Bone Active Portion
derived from a nitric oxide agent. In some embodiments, R.sub.A can
be a Bone Active Portion derived from the nitric oxide agent,
alkoxy-NO.sub.2, as represented by the following formula, where D
is
##STR00228##
and E and G are covalent bonds:
##STR00229##
The Bone Active Portion derived from alkoxy-NO.sub.2 is
alkoxy-NO.sub.2 less a hydrogen, allowing the Bone Active Portion
to be connected to the remainder of the compound, while maintaining
one or more activities generally associated with free
alkoxy-NO.sub.2.
[0274] Another exemplary compound of the presently-disclosed
subject matter can be represented by the following formula:
##STR00230##
where the Linking Portion is connected to the Bone Targeting
Portion (R.sub.T) at R.sub.1; where: D is
##STR00231##
and E and G are covalent bonds; m is 1; n is 2; each R.sub.S is
hydrogen; R.sub.2, R.sub.3, R.sub.5, R.sub.6 are each H; R.sub.4 is
methyl; R.sub.7 is amino; and where R.sub.A is derived from
alkoxy-NO.sub.2.
[0275] In some embodiments, R.sub.A can be a Bone Active Portion
derived from an androgen. In some embodiments, R.sub.A can be a
Bone Active Portion derived from the androgen,
##STR00232##
[0276] DHEA, as represented by the following formula, where D is E
is --(CH.sub.2).sub.2--, and G is a covalent bond:
##STR00233##
The Bone Active Portion derived from DHEA is DHEA singly bonded to
oxygen at carbon 17, allowing the Bone Active Portion to be
connected to the remainder of the compound, while maintaining one
or more activities generally associated with free DHEA.
[0277] Another exemplary compound of the presently-disclosed
subject matter can be represented by the following formula:
##STR00234##
where the Linking Portion is connected to the Bone Targeting
Portion (R.sub.T) at R.sub.1; where: D is
##STR00235##
E is --(CH.sub.2).sub.2--, and G is a covalent bond; m is 1; n is
2; each R.sub.S is hydrogen; R.sub.2, R.sub.3, R.sub.5, R.sub.6 are
each H; R.sub.4 is methyl; R.sub.7 is amino; and where R.sub.A is
derived from DHEA.
[0278] In some embodiments, R.sub.A can be a Bone Active Portion
derived from the androgen, testosterone, as represented by the
following formula, where D is
##STR00236##
E is --(CH.sub.2).sub.2--, and G is a covalent bond:
##STR00237##
The Bone Active Portion derived from testosterone can be
testosterone less a hydrogen, allowing the Bone Active Portion to
be connected to the remainder of the compound, while maintaining
one or more activities generally associated with free
testosterone.
[0279] Another exemplary compound of the presently-disclosed
subject matter can be represented by the following formula:
##STR00238##
where the Linking Portion is connected to the Bone Targeting
Portion (R.sub.T) at R.sub.1; where: D is
##STR00239##
E is --(CH.sub.2).sub.2--; G is a covalent bond; m is 1; n is 2;
each R.sub.S is hydrogen; R.sub.2, R.sub.3, R.sub.5, R.sub.6 are
each H; R.sub.4 is methyl; R.sub.7 is amino; and where R.sub.A is
derived from testosterone.
[0280] In some embodiments, R.sub.A can be a Bone Active Portion
derived from a carbonic anhydrase inhibitor. In some embodiments,
R.sub.A can be a Bone Active Portion derived from the carbonic
anhydrase inhibitor, 2-aminothiadiazole-5-sulfonamide, as
represented by the following formula, where D and G are
##STR00240##
and E is --(CH.sub.2).sub.3)--:
##STR00241##
[0281] The Bone Active Portion derived from
2-amino-1,3,4-thiadiazole-5-sulfonamide is 2-amino-1,
3,4-thiadiazole-5-sulfonamide less a hydrogen, allowing the Bone
Active Portion to be connected to the remainder of the compound,
while maintaining one or more activities generally associated with
free sulfonamide.
[0282] Another exemplary compound of the presently-disclosed
subject matter can be represented by the following formula:
##STR00242##
where the Linking Portion is connected to the Bone Targeting
Portion (R.sub.T) at R.sub.1; where: D and G are
##STR00243##
and E is --(CH.sub.2).sub.3--; m is 1; n is 2; each R.sub.S is
hydrogen; R.sub.2, R.sub.3, R.sub.5, R.sub.6 are each H; R.sub.4 is
methyl; R.sub.7 is amino; and where R.sub.A is derived from
2-aminothiadiazole-5-sulfonamide.
[0283] In some embodiments, R.sub.A can be a Bone Active Portion
derived from an anti-cancer agent or an antineoplastic agent. In
some embodiments, R.sub.A can be a Bone Active Portion derived from
doxorubicin, as represented by the following formula, where D and G
are
##STR00244##
and E is --(CH.sub.2).sub.3--:
##STR00245##
[0284] The Bone Active Portion derived from doxorubicin is
doxorubicin less a hydrogen, allowing the Bone Active Portion to be
connected to the remainder of the compound, while maintaining one
or more activities generally associated with free doxorubicin.
[0285] Another exemplary compound of the presently-disclosed
subject matter can be represented by the following formula:
##STR00246##
where the Linking Portion is connected to the Bone Targeting
Portion (R.sub.T) at R.sub.1; where: D and G are
##STR00247##
E is --(CH.sub.2).sub.3--; m is 1; n is 2; each R.sub.S is
hydrogen; R.sub.2, R.sub.3, R.sub.5, R.sub.6 are each H; R.sub.4 is
methyl; R.sub.7 is amino; and where R.sub.A is derived from
doxorubicin.
[0286] In some embodiments, R.sub.A can be a Bone Active Portion
derived from dasatinib, as represented by the following formula,
where D is
##STR00248##
E is --(CH.sub.2)--, and G is covalent bond:
##STR00249##
The Bone Active Portion derived from dasatinib is dasatinib less a
hydrogen, allowing the Bone Active Portion to be connected to the
remainder of the compound, while maintaining one or more activities
generally associated with free dasatinib.
[0287] Another exemplary compound of the presently-disclosed
subject matter can be represented by the following formulas:
##STR00250##
where the Linking Portion is connected to the Bone Targeting
Portion (R.sub.T) at R.sub.1; where: D is
##STR00251##
E is --(CH.sub.2)--; G is covalent bond; m is 1; n is 2; each
R.sub.S is hydrogen; R.sub.2, R.sub.3, R.sub.5, R.sub.6 are each H;
R.sub.4 is methyl; R.sub.7 is amino; and where R.sub.A is derived
from dasatinib.
[0288] In some embodiments, R.sub.A can be a Bone Active Portion
derived from an antimicrobial agent. In some embodiments, R.sub.A
can be a Bone Active Portion derived from the antimicrobial agent,
vancomycin, as represented by the following formula, where D is
##STR00252##
and E and G are a covalent bond:
##STR00253##
The Bone Active Portion derived from vancomycin can be vancomycin
less a hydroxyl, allowing the Bone Active Portion to be connected
to the remainder of the compound, while maintaining one or more
activities generally associated with free vancomycin.
[0289] Another exemplary compound of the presently-disclosed
subject matter can be represented by the following formula:
##STR00254##
where the Linking Portion is connected to the Bone Targeting
Portion (R.sub.T) at R.sub.1; where: D is
##STR00255##
E and G are a covalent bond; m is 1; n is 2; each R.sub.S is
hydrogen; R.sub.2, R.sub.3, R.sub.5, R.sub.6 are each H; R.sub.4 is
methyl; R.sub.7 is amino; and where R.sub.A is derived from
vancomycin.
[0290] In some embodiments of the compounds of the
presently-disclosed subject matter, the Linker Portion of Formula
12 is used and the Linker Portion is connected to the Bone
Targeting Portion at R.sub.4, as represented by the following
formula:
##STR00256##
Because the -D-E-G- segment is positioned adjacent the third unit,
R.sub.A, it is beneficial to select -D-E-G-based on the selected
third unit, R.sub.A. For example, when R.sub.A is a Bone Active
Portion, it is contemplated that the Linker Portion can be bound to
the Bone Active Portion to minimize the susceptibility to
hydrolysis, e.g., ester, urido, ether, linkage, to increase the bio
availability of the compound. That is to say, if susceptibility to
hydrolysis is minimized, without wishing to be bound by theory or
mechanism, the compound can be delivered to and affect bone.
[0291] As mentioned above, the third unit R.sub.A of the compound
can be selected from: a protecting group; a hydrogen when the
Linking Portion of Formula 12 is selected; or a Bone Targeting
Portion (See e.g., Tables A-D).
[0292] When the protecting group t-butoxycarbonyl (t-BOC) is used,
as will be understood by those skilled in the art, it can be
beneficial in some embodiments for D, E, and G to be a covalent
bond. In this regard, an exemplary compound of the
presently-disclosed subject matter can be represented by the
following formula:
##STR00257##
which is the exemplary compound of Formula 152, where D, E, and G
are covalent bonds, and R.sub.A is t-BOC.
[0293] Another exemplary compound of the presently-disclosed
subject matter can be represented by the following formula:
##STR00258##
which is the exemplary compound of Formula 152, where D, E, and G
are a covalent bond, and R.sub.A is H.
[0294] In some embodiments, R.sub.A can be derived from a bone
active agent. In some embodiments, R.sub.A can be a Bone Active
Portion derived from a steroidal estrogenic agent. In some
embodiments, R.sub.A can be a Bone Active Portion derived from the
a steroidal estrogenic agent, estradiol. As will be understood by
those skilled in the art, in such cases, it can be beneficial in
some embodiments for D to be
##STR00259##
for E to be a group other than a covalent bond, and for G to be a
covalent bond. In this regard, in some embodiments, the compound
can be represented by the following formula, where D is
##STR00260##
E is --(CH.sub.2).sub.2--, and G is a covalent bond, and R.sub.A is
derived from estradiol:
##STR00261##
[0295] The Bone Active Portion derived from estradiol is estradiol
less a hydrogen, allowing the Bone Active Portion to be connected
to the remainder of the compound, while maintaining one or more
activities generally associated with free estradiol. In some
embodiments, when the Bone Active Portion of the compound is
derived from estradiol, it is derived from the 17-.beta.-enantiomer
of estradiol. Without wishing to be bound by theory or mechanism,
it is believed that the 17-.beta.-enantiomer of estradiol is the
active isomer.
[0296] Another exemplary compound of the presently-disclosed
subject matter can be represented by the following formula:
##STR00262##
where D is
##STR00263##
E is --(CH.sub.2).sub.2--; G is a covalent bond; m is 1; n is 2;
each R.sub.S is H; R.sub.1, R.sub.2, R.sub.3, R.sub.5, and R.sub.6
are each H; R.sub.7 is NO.sub.2; and where R.sub.A is derived from
estradiol.
[0297] In some embodiments, R.sub.A can be a Bone Active Portion
derived from a non-steroidal estrogenic agent. In some embodiments,
R.sub.A can be a Bone Active Portion derived from the non-steroidal
estrogenic agent, genistein, as represented by the following
formula
##STR00264##
The Bone Active Portion derived from genistein is genistein less a
hydrogen, allowing the Bone Active Portion to be connected to the
remainder of the compound, while maintaining one or more activities
generally associated with free genistein.
[0298] Another exemplary compound of the presently-disclosed
subject matter can be represented by the following formula:
##STR00265##
where the Linking Portion is connected to the Bone Targeting
Portion (R.sub.T) at R.sub.4; where: D is
##STR00266##
E is --(CH.sub.2)--; G is a covalent bond; m is 1; n is 2; each
R.sub.S is hydrogen; R.sub.1, R.sub.2, R.sub.3, R.sub.5, and
R.sub.6 are each H; R.sub.7 is NO.sub.2; and where R.sub.A is
derived from genistein.
[0299] In some embodiments, R.sub.A can be a Bone Active Portion
derived from a nitric oxide agent. In some embodiments, R.sub.A can
be a Bone Active Portion derived from the nitric oxide agent,
alkoxy-NO.sub.2, as represented by the following formula, where D
is
##STR00267##
and E and G are covalent bonds:
##STR00268##
The Bone Active Portion derived from alkoxy-NO.sub.2 is
alkoxy-NO.sub.2 less a hydrogen, allowing the Bone Active Portion
to be connected to the remainder of the compound, while maintaining
one or more activities generally associated with free
alkoxy-NO.sub.2.
[0300] Another exemplary compound of the presently-disclosed
subject matter can be represented by the following formula:
##STR00269##
where the Linking Portion is connected to the Bone Targeting
Portion (R.sub.T) at R.sub.4; where: D is
##STR00270##
and E and G are covalent bonds; m is 1; n is 2; each R.sub.S is
hydrogen; R.sub.1, R.sub.2, R.sub.3, R.sub.5, and R.sub.6 are each
H; R.sub.7 is NO.sub.2; and where R.sub.A is derived from
alkoxy-NO.sub.2.
[0301] In some embodiments, R.sub.A can be a Bone Active Portion
derived from an androgen. In some embodiments, R.sub.A can be a
Bone Active Portion derived from the androgen, DHEA, as represented
by the following formula, where D is
##STR00271##
E is --(CH.sub.2).sub.2--, and G is a covalent bond:
##STR00272##
The Bone Active Portion derived from DHEA is DHEA singly bonded to
oxygen at carbon 17, allowing the Bone Active Portion to be
connected to the remainder of the compound, while maintaining one
or more activities generally associated with free DHEA.
[0302] Another exemplary compound of the presently-disclosed
subject matter can be represented by the following formula:
##STR00273##
where the Linking Portion is connected to the Bone Targeting
Portion (R.sub.T) at R.sub.4; where: D is
##STR00274##
E is --(CH.sub.2).sub.2--, and G is a covalent bond; m is 1; n is
2; each R.sub.S is hydrogen; R.sub.1, R.sub.2, R.sub.3, R.sub.5,
and R.sub.6 are each H; R.sub.7 is NO.sub.2; and where R.sub.A is
derived from DHEA.
[0303] In some embodiments, R.sub.A can be a Bone Active Portion
derived from the androgen, testosterone, as represented by the
following formula, where D is
##STR00275##
E is --(CH.sub.2).sub.2--, and G is a covalent bond:
##STR00276##
The Bone Active Portion derived from testosterone is testosterone
less a hydrogen, allowing the Bone Active Portion to be connected
to the remainder of the compound, while maintaining one or more
activities generally associated with free testosterone.
[0304] Another exemplary compound of the presently-disclosed
subject matter can be represented by the following formula:
##STR00277##
where the Linking Portion is connected to the Bone Targeting
Portion (R.sub.T) at R.sub.4; where: D is
##STR00278##
E is --(CH.sub.2).sub.2--; G is a covalent bond; m is 1; n is 2;
each R.sub.S is hydrogen; R.sub.1, R.sub.2, R.sub.3, R.sub.5, and
R.sub.6 are each H; R.sub.7 is NO.sub.2; and where R.sub.A is
derived from testosterone.
[0305] In some embodiments, R.sub.A can be a Bone Active Portion
derived from a carbonic anhydrase inhibitor. In some embodiments,
R.sub.A can be a Bone Active Portion derived from the carbonic
anhydrase inhibitor, 2-aminothiadiazole-5-sulfonamide, as
represented by the following formula, where D and G are
##STR00279##
and E is --(CH.sub.2).sub.3--:
##STR00280##
[0306] The Bone Active Portion derived from
2-amino-1,3,4-thiadiazole-5-sulfonamide is 2-amino-1,
3,4-thiadiazole-5-sulfonamide less a hydrogen, allowing the Bone
Active Portion to be connected to the remainder of the compound,
while maintaining one or more activities generally associated with
free sulfonamide.
[0307] Another exemplary compound of the presently-disclosed
subject matter can be represented by the following formula:
##STR00281##
where the Linking Portion is connected to the Bone Targeting
Portion (R.sub.T) at R.sub.4; where: D and G are
##STR00282##
and E is --(CH.sub.2).sub.3--; m is 1; n is 2; each R.sub.S is
hydrogen; R.sub.1, R.sub.2, R.sub.3, R.sub.5, and R.sub.6 are each
H; R.sub.7 is NO.sub.2; and where R.sub.A is derived from
2-aminothiadiazole-5-sulfonamide.
[0308] In some embodiments, R.sub.A can be a Bone Active Portion
derived from an anti-cancer agent or an antineoplastic agent. In
some embodiments, R.sub.A can be a Bone Active Portion derived from
doxorubicin, as represented by the following formula, where D and G
are
##STR00283##
and E is --(CH.sub.2).sub.3--:
##STR00284##
[0309] The Bone Active Portion derived from doxorubicin is
doxorubicin less a hydrogen, allowing the Bone Active Portion to be
connected to the remainder of the compound, while maintaining one
or more activities generally associated with free doxorubicin.
[0310] Another exemplary compound of the presently-disclosed
subject matter can be represented by the following formula:
##STR00285##
where the Linking Portion is connected to the Bone Targeting
Portion (R.sub.T) at R.sub.4; where: D and G are
##STR00286##
E is --(CH.sub.2).sub.3--; m is 1; n is 2; each R.sub.S is
hydrogen; R.sub.1, R.sub.2, R.sub.3, R.sub.5, and R.sub.6 are each
H; R.sub.7 is NO.sub.2; and where R.sub.A is derived from
doxorubicin.
[0311] In some embodiments, R.sub.A can be a Bone Active Portion
derived from dasatinib, as represented by the following formula,
where D is
##STR00287##
E is --(CH.sub.2)--, and G is covalent bond:
##STR00288##
The Bone Active Portion derived from dasatinib is dasatinib less a
hydrogen, allowing the Bone Active Portion to be connected to the
remainder of the compound, while maintaining one or more activities
generally associated with free dasatinib.
[0312] Another exemplary compound of the presently-disclosed
subject matter can be represented by the following formulas:
##STR00289##
[0313] where the Linking Portion is connected to the Bone Targeting
Portion (R.sub.T) at R.sub.4; where: where D is
##STR00290##
E is --(CH.sub.2)--; G is covalent bond; m is 1; n is 2; each
R.sub.S is hydrogen; R.sub.1, R.sub.2, R.sub.3, R.sub.5, and
R.sub.6 are each H; R.sub.7 is NO.sub.2; and where R.sub.A is
derived from dasatinib.
[0314] In some embodiments, R.sub.A can be a Bone Active Portion
derived from an antimicrobial agent. In some embodiments, R.sub.A
can be a Bone Active Portion derived from the antimicrobial agent,
vancomycin, as represented by the following formula, where D is
##STR00291##
and E and G are a covalent bond:
##STR00292##
The Bone Active Portion derived from vancomycin can be vancomycin
less a hydroxyl, allowing the Bone Active Portion to be connected
to the remainder of the compound, while maintaining one or more
activities generally associated with free vancomycin.
[0315] Another exemplary compound of the presently-disclosed
subject matter can be represented by the following formula:
##STR00293##
where the Linking Portion is connected to the Bone Targeting
Portion (R.sub.T) at R.sub.4; where: D is
##STR00294##
E and G are a covalent bond; m is 1; n is 2; each R.sub.S is
hydrogen; R.sub.1, R.sub.2, R.sub.3, R.sub.5, and R.sub.6 are each
H; R.sub.7 is NO.sub.2; and where R.sub.A is derived from
vancomycin.
[0316] As described above, in some embodiments the compounds of the
presently-disclosed subject matter are characterized by at least
one active portion. The Bone Targeting Portion (R.sub.T) has the
ability to bind to calcium with a tendency to accumulate in bone
and to incorporate into its crystal lattice. In some embodiments,
the compounds include an additional active portion. The Bone Active
Portion (R.sub.A) can be derived from a bone active agent, which
interacts with bone and affects bone metabolism. For example, if
the Bone Active Portion (R.sub.A) is derived from vitamin D, then
its interaction with bone tends to strengthen and increase bone
formation. On the other hand, the steroids contemplated by the
presently-disclosed subject matter exhibit bone activity and can
both inhibit bone resorption and stimulate bone formation. In
addition, the carbonic anhydrase inhibitors contemplated by the
presently-disclosed subject matter inhibit the enzyme carbonic
anhydrase, which catalyzes the reversible hydration of carbon
dioxide to carbonic acid and thus it is an inhibitor of bone
resorption. Other bone active agents contemplated by the
presently-disclosed subject matter, e.g., listed in Tables A-D,
exert their known effects in a manner relatively specific to
bone.
[0317] The performance of the compounds of the presently-disclosed
subject matter can be facilitated, first by the Bone Targeting
Portion (R.sub.T), which localizes the compound at the bone site.
Once anchored at the bone site, the Bone Active Portion (R.sub.A)
of the molecule, i.e., the bone active domain, interacts with and
affects the bone.
[0318] In some embodiments, the compounds of the
presently-disclosed subject matter can be pro-drugs. In this
regard, the compound can be formulated such that the Bone Active
Portion (R.sub.A) exhibits no initial activity; however, when
subjected to the enzymatic or hydrolytic conditions occurring at
the bone site, the Bone Active Portion (R.sub.A) will become
active.
[0319] Without wishing to be bound by theory or mechanism, it is
believed that the compounds of the presently-disclosed subject
matter interact with the calcium in the bone in the following
manner, which is described using an exemplary embodiment of the
presently-disclosed subject matter:
##STR00295##
[0320] As shown by the example, in some embodiments, three
positions of the Bone Targeting Portion (R.sub.T) can interact with
calcium, facilitating the localization of the compound to bone. In
the exemplary compound of Formula 174, the R.sub.4 group (CH.sub.3)
is not depicted as interacting with the calcium; however, it is
contemplated that the R.sub.4 group can affect the affinity for
bone. Without wishing to be bound by theory or mechanism, it is
believed that the affinity for bone can be modulated, in part, by
strategically selecting the R.sub.4 group based on its
electron-donating properties. The greater the electron donating
properties of the R.sub.4 group, the greater the affinity for bone,
i.e., a negative charge is directed from the side of the Bone
Targeting Portion having the R.sub.4 group, towards the side of the
Bone Targeting Portion thought to interact with the calcium,
thereby creating a stronger interaction between the Bone Targeting
Portion and the positively-charged calcium.
[0321] Some embodiments of compounds of the presently-disclosed
subject matter are described with reference to formulas. Some
formulas include portions depicting a particular stereoisomer of
one or more moieties of the compound. Such depicted stereoisomers
are representative of some embodiments of the compounds; however,
the formulas are intended to encompass all active stereoisomers of
the depicted compounds.
[0322] The compounds of the presently-disclosed subject matter can
in some embodiments contain one or more asymmetric carbon atoms and
can exist in racemic and optically active forms. Depending upon the
substituents, the present compounds can form addition salts as
well. All of these other forms are contemplated to be within the
scope of the presently-disclosed subject matter. The compounds of
the presently-disclosed subject matter can exist in stereoisomeric
forms and the products obtained thus can be mixtures of the
isomers.
[0323] The presently-disclosed subject matter includes methods for
treating bone conditions in a subject. Methods include
administering to the subject an effective amount of a compound of
the presently-disclosed subject matters, as described above.
[0324] As used herein, the terms treatment or treating relate to
any treatment of a bone condition of interest, including but not
limited to prophylactic treatment and therapeutic treatment As
such, the terms treatment or treating include, but are not limited
to: preventing a condition of interest or the development of a
condition of interest; inhibiting the progression of a condition of
interest; arresting or preventing the development of bone condition
of interest; reducing the severity of condition of interest;
ameliorating or relieving symptoms associated with a condition of
interest; and causing a regression of the condition of interest or
one or more of the symptoms associated with the condition of
interest. Examples of conditions of interest are noted herein. For
example, in some embodiments, the condition of interest can be a
primary or secondary bone condition of interest.
[0325] As noted above, in some embodiments, the bone condition of
interest is a metabolic bone disease (MBD), wherein treatment can
result in an anti-catabolic effect and/or an anabolic effect. In
some embodiments, the bone condition of interest is a bone
fracture, wherein treatment can result in an anabolic effect. In
some embodiments, the bone condition of interest is a bone cancer,
wherein treatment can result in an anti-cancer effect. In some
embodiments, the bone condition of interest is a bone microbial
infection, wherein treatment can result in an anti-microbial
effect. Other conditions of interest and/or desired effects are
noted herein and/or are contemplated by the presently-disclosed
subject matter.
[0326] As used herein, the term effective amount refers to a dosage
sufficient to provide treatment for the bone condition of interest
being treated. This can vary depending on the patient, the
condition, and the treatment being effected. The exact amount that
is required will vary from subject to subject, depending on the
species, age, and general condition of the subject, the particular
carrier or adjuvant being used, mode of administration, and the
like. As such, the effective amount will vary based on the
particular circumstances, and an appropriate effective amount can
be determined in a particular case by one of ordinary skill in the
art using only routine experimentation.
[0327] As noted above, in some embodiments, the compound can be
provided as a pharmaceutically-acceptable salt or solvate. Suitable
acids and/suitable bases, as will be known to those of ordinary
skill in the art, are capable of forming salts of the compounds
described herein, e.g., hydrochloric acid (HCl), sodium hydroxide.
A solvate is a complex or aggregate formed by one or more molecules
of a solute, e.g. a compound or a pharmaceutically-acceptable salt
thereof, and one or more molecules of a solvent. Such solvates can
be crystalline solids having a substantially fixed molar ratio of
solute and solvent. Suitable solvents will be known by those of
ordinary skill in the art, e.g., water, ethanol.
[0328] As will be understood by those of ordinary skill in the art,
a dosage regimen can be adjusted to provide an optimum treatment
effect and can be administered daily, biweekly, weekly, bimonthly,
monthly, or at other appropriate time intervals. As will be
understood by those of ordinary skill in the art, compounds of the
presently-disclosed subject matter can be administered orally,
intravenously, intramuscularly, subcutaneously, or by other
art-recognized means.
[0329] For oral administration, the compositions can take the form
of, for example, tablets or capsules prepared by a conventional
technique with pharmaceutically acceptable excipients such as
binding agents (e.g., pregelatinized maize starch,
polyvinylpyrrolidone or hydroxypropyl methylcellulose); fillers
(e.g., lactose, microcrystalline cellulose or calcium hydrogen
phosphate); lubricants (e.g., magnesium stearate, talc or silica);
disintegrants (e.g., potato starch or sodium starch glycollate); or
wetting agents (e.g., sodium lauryl sulphate). The tablets can be
coated by methods known in the art. Liquid preparations for oral
administration can take the form of, for example, solutions, syrups
or suspensions, or they can be presented as a dry product for
constitution with water or other suitable vehicle before use. Such
liquid preparations can be prepared by conventional techniques with
pharmaceutically acceptable additives such as suspending agents
(e.g., sorbitol syrup, cellulose derivatives or hydrogenated edible
fats); emulsifying agents (e.g. lecithin or acacia); non-aqueous
vehicles (e.g., almond oil, oily esters, ethyl alcohol or
fractionated vegetable oils); and preservatives (e.g., methyl or
propyl-p-hydroxybenzoates or sorbic acid). The preparations can
also contain buffer salts, flavoring, coloring and sweetening
agents as appropriate. Preparations for oral administration can be
suitably formulated to give controlled release of the active
compound. For buccal administration the compositions can take the
form of tablets or lozenges formulated in conventional manner.
[0330] The compounds can also be formulated as a preparation for
injection. Thus, for example, the compounds can be formulated with
a suitable carrier. The carrier can be a solvent or dispersion
medium containing, for example, water, ethanol, polyol (for
example, glycerol, propylene glycol, and liquid polyethylene
glycol, and the like), suitable mixtures thereof, and vegetable
oils. The proper fluidity can be maintained, for example, by the
use of a coating such as lecithin, by the maintenance of the
required particle size in the case of dispersion and by the use of
surfactants.
[0331] As used herein, the term subject refers to humans and other
animals. Thus, veterinary uses are provided in accordance with the
presently disclosed subject matter. The presently disclosed subject
matter provides for the treatment of mammals such as humans, as
well as those mammals of importance due to being endangered, such
as Siberian tigers; of economic importance, such as animals raised
on farms for consumption by humans; and/or animals of social
importance to humans, such as animals kept as pets or in zoos.
Examples of such animals include but are not limited to: carnivores
such as cats and dogs; swine, including pigs, hogs, and wild boars;
ruminants and/or ungulates such as cattle, oxen, sheep, giraffes,
deer, goats, bison, and camels; and horses. Also provided is the
treatment of birds, including the treatment of those kinds of birds
that are endangered and/or kept in zoos, as well as fowl, and more
particularly domesticated fowl, i.e., poultry, such as turkeys,
chickens, ducks, geese, guinea fowl, and the like, as they are also
of economic importance to humans. Thus, also provided is the
treatment of livestock, including, but not limited to, domesticated
swine, ruminants, ungulates, horses (including race horses),
poultry, and the like.
[0332] The presently-disclosed subject matter includes a method of
making a bone-targeted compound for the treatment of a bone
condition of interest. A bone active agent is selected, from which
the bone active portion of the compound will be derived. An
appropriate stably-stored compound of the presently-disclosed
subject matter is selected, wherein R.sub.A is a protecting group,
hydrogen, or hydroxyl, as described above. The stably-stored
compound and the bone active agent are used to prepare a compound,
wherein R.sub.A is a bone active portion derived from the selected
bone active agent. In some embodiments, the selected bone active
agent has an independent ability to treat the bone condition of
interest; however, the resulting compound having the bone active
portion derived from the bone active agent has an enhanced ability
to target bone and treat the bone condition of interest, and a
reduced capacity for negative side effects. In this regard, the
methods and compounds of the presently-disclosed subject matter can
be used to salvage once-promising treatment compounds that were
abandoned due to insufficient bioavailability, insufficient
bioactivity, and/or unacceptable side effects.
[0333] The presently-disclosed subject matter is further
illustrated by the following specific but non-limiting examples.
The following examples may include compilations of data that are
representative of data gathered at various times during the course
of development and experimentation related to the
presently-disclosed subject matter. The following examples include
some examples that are prophetic.
EXAMPLES
[0334] Syntheses. The compounds of the presently-disclosed subject
matter can be prepared in accordance with the exemplary schemes set
forth in the Examples herein, and by techniques known to those of
ordinary skill in the art.
Exemplary Synthesis where R.sub.A is a Protecting Group
[0335] The following is exemplary, where R.sub.A is the Protecting
Group, FMOC. About 2.5 g of FMOC-8-amino-3,6-dioxaoctanoic acid
(Peptides International, Inc., Louisville, Ky.), about 0.9 g of
hydroxybenzatriazol (HOBt), about 2.9 g of
benzotriazole-1-yl-oxy-tris-(dimethylamino)-phosphonium
hyexafluorophosphate (BOP), and about 1.1 ml diisopropylethylamine
(DIEA) are dissolved in about 20 ml dimethylformamide (DMF). In a
separate flask, about 0.89 g of
2-hydroxy-6-methoxy-3-amino-benzamide is dissolved in about 20 ml
DMF. The two solutions are mixed together and stirred at room
temperature for about 24 hours. Thin layer chromatography shows no
indication of remaining starting material. The DMF is removed under
vacuum, and crude product appears as a brown oil. The crude product
is take up in ethyl acetate, and washed with brine 2.times., 5% HCl
2.times., and saturated NaHCO.sub.3 2.times., the dried over sodium
sulfate. The product is then condensed via rotary evaporation,
resulting in a sticky brown oil. When the resulting oil is
dissolved in methanol and triturated, a brownish-white solid
precipitates. The mixture is stored at about 35.degree. F. The
precipitate is collected via vacuum filtration. The yield is
approximately 2 g, about 75%. .sup.1H NMR data: .delta.3.15, m, 2H,
6' CH.sub.2, .delta.3.44, t, J=5.98 Hz, 2H, 5' CH.sub.2,
.delta.3.60, br s, 2H, 4' CH.sub.2, .delta.3.69, br s, 2H, 3'
CH.sub.2, .delta.3.88, s, 3H, C6 OCH.sub.3, .delta.4.09, s, 2H, 2'
CH.sub.2, .delta.4.19, q, J=6.59 Hz, 1H, 9' CH, .delta.4.27, d,
J=6.40 Hz, 2H, 8' CH.sub.2, .delta.6.54, d, J=8.97 Hz, 1H, C5H,
.delta.7.31, m, 3H, C14' H/C6 amide H, .delta.7.40, m, 2H, C12' H,
.delta.7.67, d, J=7.87 Hz, 2H, C13' H, .delta.7.88, d, J=7.32 Hz,
C11' H, .delta.8.18, d, J=8.60 Hz, 1H, C4H, .delta.8.29, s, 1H, Cl
amide H, .delta.8.33, s, 1H, Cl amide H, .delta.8.85, s, 1H, C3
amide H.
##STR00296##
[0336] The following are exemplary for deprotection of compounds
where R.sub.A is FMOC, where HCl salts are formed. About 1.8 g of
FMOC-8-amino-3,6-dioxaoctanoic acid-BTA is dissolved in 20%
piperidine in DMF and stirred for about 20 minutes. The solvent is
removed and fresh piperidine solution is added to repeat the
deprotection reaction. Deprotection is performed about three times.
After the final solvent removal, a solid product appears and
diethyl ether is added. The mixture is acidified with 7N HCl in
methanol and decanted. After triturating in dichloromethane, an
off-white precipitate is collected via vacuum filtration. The yield
is about 0.94 g, about 86%. .sup.1H, .delta.2.96, t, 2H, 6'
CH.sub.2, .delta.3.65, m, 4H, 4'/5' CH.sub.2, .delta.3.73, m, 2H,
3' CH.sub.3, .delta.3.89, s, 3H, C6 OCH.sub.3, .delta.4.12, 2 H, 2'
CH.sub.2, .delta.6.56, d, J=8.91 Hz, 1H, C5H, .delta.8.17, d,
J=9.15 Hz, 1H, C4H, .delta.8.30, s, 1H, Cl amide H, .delta.8.35, s,
1H, Cl amide H, .delta.8.87, s, 1H, C3 amide H, .sup.13C,
.delta.39.22; C6', .delta.57.03, C6-OCH.sub.3, .delta.67.34, C5',
.delta.70.46, C4', .delta.70.93, C3', .delta.85.93, C2',
.delta.99.21, C6, .delta.156.70, C1', .delta.159.83, C4.
##STR00297##
[0337] The procedures described above were used for the following
transformations as well:
##STR00298##
[0338] The following is exemplary, where R.sub.A is t-BOC:
##STR00299##
Exemplary Synthesis where R.sub.A is Derived from a Steroidal
Estrogenic Agent
[0339] The following is exemplary, where R.sub.A is a Bone Active
Portion derived from Estradiol (compound of Formula 131), and the
starting material is a compound of the presently disclosed subject
matter, where R.sub.A is a protecting group.
##STR00300##
[0340] The following is another exemplary manner in which the
compound of Formula 131 can be prepared.
##STR00301##
[0341] The following is exemplary for preparation of the compound
of Formula 131. In a round-bottom flask about 0.48 g (1.4 mmol)
17-.beta.-estro-O-propanoic acid, about 0.38 g (2.8 mmol)
hydroxybenzotriazole (HOBt), about 1.24 g (2.8 mmol)
benzotriazole-1-yl-oxy-tris-(dimethylamino)-phosphonium
hexafluorophosphate (BOP), and about 0.5 g (1.4 mmol) NBR-VI-232-1
(synthesis described herein) were combined. The dry mixture was
dissolved in about 20 mL dimethylformamide (DMF) then about 488
.mu.L (2.8 mmol) of N,N-diispropylethylamine (DIEA) was added. The
reaction was allowed to stir at room temperature for about 48
hours.
[0342] The reaction solvent was removed under vacuum and the
resulting yellow oil was taken up in about 50 mL ethylacetate
(EtOAc). It was then washed with brine 2.times., 5% HC12x, and
saturated sodium bicarbonate 2.times. and dried over sodium
sulfate. After gravity filtration, the filtrate was condensed via
rotary evaporation to yield a sticky oil. Upon trituration of the
oil in cold methanol, product NBR-VI-240-1 precipitated out as an
off-white solid. The product was collected via vacuum filtration.
Yield: 510 mg, 55.7%. .sup.1H, .delta.0.66, s, 3H, angular methyl,
.delta.1.10, m, .delta.1.16-1.32, m, .delta.1.35, m, .delta.1.55,
m, .delta.1.73, m, .delta.1.86-1.97, m, .delta.2.04, t,
.delta.2.19, d, .delta.2.27, t, J 6.47 Hz, 2H, 8' CH.sub.2,
.delta.2.68, m, 2H, .delta.3.21, q, J=5.61 Hz, 2H, 6' CH.sub.2,
.delta.3.44, t, J=5.86 Hz, 2H, 5' CH.sub.2, .delta.3.55, m, 1H, C9'
H, .delta.3.60, m, 2H, 4' CH.sub.2, .delta.3.69, m, 2H, 3'
CH.sub.2, .delta.3.63, m, 1H, C9' H.sub.2, .delta.3.87, s, 3H, C6
OCH.sub.3, .delta.4.09, s, 2H, 2' CH.sub.2, .delta.6.42, br s, 1H,
C18' H, .delta.6.49, dd, J.sub.1 8.54 Hz, J.sub.2 2.20 Hz, 1H, C20'
H, .delta.6.54, d, J=9.03 Hz, 1H, C5H, .delta.7.01, J=8.54 Hz, 1H,
C21' H, .delta.7.81, t, J=5.61 Hz, 1H, C6' amide H, .delta.8.19, d,
J=9.03 Hz, 1H, C4H, .delta.8.29, s, 1H, Cl amide H, .delta.8.33, s,
1H, Cl amide H, .delta.8.85, s, 1H, C3 amide H, .delta.8.96, s, 1H,
C19' OH. Thin-Layer Chromatography: R.sub.f=0.62 in
EtOAc:Acetone:Methanol, 7:7:5.
##STR00302##
[0343] The compound of Formula 132 can be prepared in the following
manner.
##STR00303##
[0344] The compound of Formula 131 was formed as described in
above. The product was oily, but after sitting in hexane at
4.degree. C., a light gray precipitate could be collected via
vacuum filtration. This solid was subjected to thin-layer
chromatography and .sup.1H NMR for characterization. TLC:
R.sub.f=0.64 in EtOAc:Acetone:MeOH (18:2:5) and a 1-dimensional
.sup.1H NMR spectrum confirms the structure. The compound begins
decomposing at 93.degree. C. and melts from 97-100.degree.. The
yield was about 50 mg, 14.4%.
Exemplary Synthesis where R.sub.A is a Protecting Group
[0345] A exemplary compound of the presently-disclosed subject
matter can be prepared in the following manner.
##STR00304##
[0346] In a 200 mL round-bottom flask were dissolved 5.00 g (13
mmol) Fmoc-MiniPEG, 2.95 g (10 mmol) of
2-hydroxy-6-benzyloxy-3-amino-benzamide hydrochloride (BTA RIC-T3),
3.51 g (26 mmol) HOBt, 11.5 (26 mmol) BOP, and 4.53 mL (26 mmol)
DIEA, in 70 mL of DMF. The homogeneous solution was allowed to stir
at room temperature overnight.
[0347] The DMF was removed under vacuum after .about.22 hours to
give a dark brown oil. The oil was dissolved in 100 mL EtOAc and
washed with brine 2.times., 5% HCl 2.times., and saturated
NaHCO.sub.3 2.times., then dried over sodium sulfate and
concentrated via rotary evaporation to give a sticky brown residue.
The residue was taken up in methanol and triturated. An off-white
precipitate (NBR-VI-227-2) was collected and washed with MeOH via
vacuum filtration. The yield was about 4.67 g, 70%. .sup.1H,
.delta.3.15, t, J=5.50 Hz, 2H, 6' CH.sub.2, .delta.3.44, t, J=5.73
Hz, 2H, 5' CH.sub.2, .delta.3.59, br s, 2H, 4' CH.sub.2,
.delta.3.67, br s, 2H, 3' CH.sub.2, .delta.4.19, t, J=6.71 Hz, 1H,
C9' H, .delta.4.27, d, J=6.83 Hz, 2H, 2' CH.sub.2, .delta.5.27, s,
2H, C6 CH.sub.2 (benzyl), .delta.6.64, d, J 8.78H, 1H, C5H,
.delta.7.31, m, 3H, C14' H/benzyl H.sub.c, .delta.7.36, d, J=7.32
Hz, 1H, 6' amide H, .delta.7.40, m, 4H, C12' H/benzyl H.sub.b,
.delta.7.48, d, J=7.20 Hz, 2H, benzyl H.sub.a, .delta.7.67, d,
J=7.20 Hz, 2H, C13' H, .delta.7.87, d, J=7.69 Hz, 2H, C11' H,
.delta.8.13 Hz, d, J=4.52 Hz, 1H, C4H, .delta.8.19, s, 1H, Cl amide
H, .delta.8.36, s, 1H, Cl amide H, .delta.8.86, s, 1H, C3 amide
H.
[0348] The following are exemplary for deprotection of compounds
where R.sub.A is FMOC, where HCl salts are formed. About 2.50 g
(3.8 mmol) of NBR-VI-227-2 was dissolved and stirred in about 20 mL
of 20% piperidine in DMF for 20 minutes before the solvent was
removed under vacuum. The piperidine treatment and solvent removal
were repeated twice more. After the final solvent removal, the
mixture was taken up in ether and acidified with 7N HCl in MeOH. A
white precipitate was collected via filtration, however the solid
proved to be the Fmoc hydrocarbon. The remaining filtrate was
concentrated and taken up in a small amount of methanol. It was
then allowed to sit in MeOH at 4.degree. C. for 48 hours. A
solid/oil mixture resulted. The mixture was dried under vacuum for
1 hour, taken up in ether, then briefly exposed to dry ice-acetone
bath and triturated. After some 10 minutes, a light purple
precipitate was noted. This solid, NBR-VI-236-1, was collected via
vacuum filtration. It was then recrystallized from methanol/ether
to give off-white solid, NBR-VI-236-2. The yield was about 740 mg,
56.5%. .sup.1H, .delta.2.94, t, J=5.37 Hz, 2H, 6' CH.sub.2,
.delta.3.63, m, 4H, 4'/5' CH.sub.2, .delta.3.71, m, 3' CH.sub.2,
.delta.4.11, s, 2H, 2' CH.sub.2, .delta.5.29, s, 2H, benzyl
CH.sub.2, .delta.6.65, d, J=9.15 Hz, 1H, C5H, .delta.7.36, d,
J=7.20 Hz, 1H, benzyl H.sub.c, .delta.7.41, t, J=7.44 Hz, 2H,
benzyl H.sub.b, .delta.7.49, d, J=7.69 Hz, 2H, benzyl H.sub.a,
.delta.8.12, d, J=9.03 Hz, 1H, C4H, .delta.8.20, s, 1H, Cl amide H,
.delta.8.37, s, 1H, Cl amide H, .delta.8.88, s, 1H, C3 amide H.
##STR00305##
Exemplary Synthesis where R.sub.A is Derived from a Steroidal
Estrogenic Agent
[0349] The compound of Formula 133 (product NBR-VI-247-2) can be
prepared in the following manner.
##STR00306##
[0350] In a 25 mL round-bottom flask was dissolved 300 mg (0.7
mmol) NBR-VI-236-2, 250 mg (0.7 mmol) .beta.-estro-propanoic acid,
620 mg (1.4 mmol) BOP, 189 mg (1.4 mmol) HOBt, and 244 .mu.l DIEA
in 15 mL of DMF. The reaction was allowed to stir at room
temperature for two days, after which time the solvent was removed
under vacuum. The resulting oil was taken up in ethyl acetate and
washed with brine 2.times., 5% HCl 2.times., and saturated
NaHCO.sub.3 2.times.. The organic layer was then dried over
Na.sub.sSO.sub.4, filtered, and concentrated. Product NBR-VI-2471
resulted as an oil. The product was combined with another oily
product (NBR-VI-246-1) obtained from a previous experiment, then
subjected to column chromatography (solvent system=EtOAc:Acetone,
9:1). After the appropriate fractions were pooled and concentrated,
sticky gray oil NBR-VI-247-2 was obtained. The yield was about 365
mg, 71.4%
[0351] The following is exemplary, where R.sub.A is a Bone Active
Portion derived from Estradiol.
##STR00307##
Exemplary Synthesis where R.sub.A is Derived from a Nonsteroidal
Estrogenic Agent
##STR00308##
[0353] The compound wherein R.sub.A was derived from genistein was
prepared as follows. Genistein (500 mg, 1.8 mmol) was stirred in 50
mL of DMF along with potassium t-butoxide (438 mg, 3.9 mmol) at
ambient temperature under nitrogen for 3 hours. The reaction
solution was then cooled to -45.degree. C. in a dry
ice-acetonitrile bath and t-butyl bromoacetate (274 .mu.L, 1.8
mmol) was added. The reaction was allowed to stir for 18 hours in
the dry ice bath, slowly warming to room temperature. The solvent
was then removed under vacuum and the resulting residue was taken
up in water and acidified with 5% HCl (aq). An off-white
precipitate was collected via vacuum filtration. Both thin-layer
chromatography and proton NMR were conducted to confirm that
desired protect had been prepared. The off-white precipitate
product was used in the following step.
##STR00309##
[0354] The off-white precipitate product produced as described in
the preceding paragraph (100 mg, 0.3 mmol) was stirred in 10 mL of
dichloromethane (DCM) and 3 mL trifluoroacetic acid for 45 minutes
at room temperature. The solvent was then removed in vacuo and the
residue was taken up in DCM and evaporated to dryness three times
to yield a light yellow product.
[0355] The bright yellow product (100 mg (est.), 0.3 mmol) was
stirred in 5 mL DCM along with diisopropylethylamine (DIEA; 157
.mu.L, 0.9 mmol) and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide
(EDC; 115 mg, 0.6 mmol). After 5 minutes, a bone-targeted miniPEG
compound (109 mg, 0.3 mmol) was added. DMF (7 ml) was added to
assist with solubilizing the miniPEG compound. After two hours, the
reaction mixture was filtered and the filtrate washed with brine
2.times., 5% HCl 2.times., and saterated sodium bicarbonate
2.times., then dried over sodium sulfate. The organic solvent was
removed under vacuum and the resulting white residue was
characterized with TLC and .sup.1H NMR, which indicated that the
desired product had been made.
Exemplary Synthesis where R.sub.A is Derived from a Nitric-Oxide
Agent
##STR00310##
[0357] Hex-5-enoic acid methyl ester was prepared as follows.
Hex-5-enoic acid (2.00 g, 17.5 mmol) was dissolved in diethyl ether
(8 mL) and methanol (4 mL) and cooled to 0.degree. C. To this
solution was added 2.0 M Trimethylsilyl diazomethane (9 mL)
dropwise. It was noted that a yellow TMS-diazomethane solution
dissipates upon stirring with the reaction mixture, and the
reaction is deemed complete when the yellow color persists. Caution
should be observed, as the reaction is associated with vigorous gas
evolution. When the yellow color persists in the reaction mixture,
TLC analysis was conducted and confirmed consumption of the acid.
The solvent was evaporated under reduced pressure carefully
(product is somewhat volatile) and replaced with ethyl acetate. The
solution was washed with saturated NaHCO.sub.3, dried, and the
filtrate was used directly in the next step. .sup.1H NMR (400 MHz)
CDCl.sub.3 .delta. 5.45 (m, 1H), .delta. 4.9 (m, 2H), .delta. 3.65
(s, 3H), .delta. 2.20 (t, 2H), .delta. 2.05 (m, 2H), .delta. 1.70
(m, 2H).
[0358] 5,6 dinitroxyhexanoic acid was prepared as follows. A 0.37 M
solution of hex-5-enoic acid methyl ester in ethyl acetate (30 mL,
11.1 mmol) was treated with Br.sub.2 (1.78 g, 11.1 mmol) dropwise.
The color dissipated after stirring and upon full addition a light
yellow color resulted. After stirring for 1 h at room temperature,
the solvent was switched to acetonitrile (40 mL) and AgNO.sub.3
(7.5 g, 44.4 mmol) was added in one portion. The heterogeneous
mixture was refluxed for 12 h, cooled to rt and filtered. The
filtrate was concentrated and switched with ethyl acetate so that
it could be washed with water three times (40 mL). The organic
layer was dried, concentrated and used directly in the next step.
The crude yellow oil (assume 11.1 mmol) was treated with THF (3 mL)
and water (1 mL) and cooled to 0.degree. C. LiOH monohydrate (0.50
g, 12 mmol) was added in one portion and stirred at 0.degree. C.
for 1 hour and allowed to warm to rt. TLC confirmed the
disappearance of SM. The reaction mixture was concentrated to
remove THF, ethyl acetate and 0.25M KHSO.sub.4 was added. The
organic layer was dried and concentrated to provide a light brown
oil. The key structural components of the compound were confirmed
and the material was used in the following step.) .sup.1H NMR (400
MHz) CDCl3 .delta. 5.35 (m AMX like, 1H), .delta. 4.65 (m AMX like,
1H), .delta. 4.40 (m AMX like, 1H), .delta. 2.40 (m, 2H), .delta.
2.05 (m, 4).
[0359] The dinitroxy compound was prepared as follows. The crude
acid from above (about 11.1 mmol) was treated with
N,Ndiisopropylethylamine (3.8 ml, 2.2 mmol), BT-2-(peg2)-NH.sub.3Cl
(4.04 g, 11.1 mmol), and DMF (20 mL). The solution was cooled to
0.degree. C. and to this solution was added HATU (4.2 g, 11.1
mmol). After stirring for 12 h, the reaction was deemed complete by
LC/MS. The reaction was added to a separatory funnel already
charged with 100 mL of EtOAc and 100 mL of water. The layers were
separated and the organic layer was washed one time with saturated
NaHCO.sub.3, followed by 10% citric acid. The organic layer was
then dried with MgSO.sub.4, filtered and concentrated. The crude
product was purified by preparative HPLC to provide 0.26 g of pure
product.
Exemplary Synthesis where R.sub.A is Derived from an Androgen
##STR00311##
[0361] The compound wherein R.sub.A was derived from testosterone
was prepared as follows. Four grams of
testosterone-17-hemisuccinate was dissolved in 25 mL
dimethylformanide (DMF) and 1.4512 g of N-hydroxybenzenetriazine
(HOBT) was added. The mixture was cooled to 0-5.degree. C. and was
stirred for 30 minutes. Diisopropylcarbodiimide (1.354 g) was
added, followed by stirring for 30 minutes at 0-5.degree. C. A
solution of 1.955 g of 3-amino-2-hydroxy-6-methyoxybenzamide in 10
mL DMF was added and the mixture was stirred for 24 hours at room
temperature. The reaction was stopped by addition of 200 mL
H.sub.2O and the mixture was extracted with 3.times.100 mL portions
of ethyl acetate which was then washed with H.sub.2O, then
2.times.25 mL portions of 10% aqueous NaHCO.sub.3, then with
H.sub.2O again. The ethyl acetate solution was dried over
Na.sub.2SO.sub.4 and concentrated under reduced pressure.
Exemplary Synthesis where R.sub.A is Derived from a Carbonic
Anhydrase Inhibitor
##STR00312##
[0363] The compound wherein R.sub.A was derived from
2-amino-1,3,4-thiadiazole-5-sulfonamide was prepared as follows.
2-amino-1,3,4-thiadiazole-5-sulfonamide is reacted with succinic
anhydride to form the succinamide derivative,
2-(4-carboxypropionylamino) 1,3,4-thiadiazole-5-sulfonamide. This
product is reacted with 3-amino-2-hydroxy-6-methoxybenzamide in the
presence of diisopropylcarbodiimide to form the above-identified
compound.
Exemplary Synthesis where R.sub.A is Derived from a Steroid
[0364] Compounds of the presently-disclosed subject matter
including a Bone Active Portion derived from a steroid can be
prepared in the following manner. A steroid of interest is
identified. Steroids can include a substituted carbon ring system
of 18 to 27 carbons, which can be saturated or partially
unsaturated. With reference to the following steroid carbon ring
skeleton, the ring system can be substituted at carbons 3, 10, 13,
and 17 with pendant heteroatoms and/or branched chain
hydrocarbons.
##STR00313##
[0365] The steroid of interest can include at least one
substitution at a carbon selected from 3, 10, 13, and 17. In some
embodiments, the at least one substitution is at a carbon selected
from 3 and 17. The at least one substitution can include between
about 2 and about 20 atoms, having a terminal group such as an
amino (--NH), hydroxyl (--OH), carboxyl (--COOH), sulfhydryl
(--SH), or another group having an active hydrogen that will allow
the steroid to undergo a condensation reaction with a compound of
the presently-disclosed subject matter where R.sub.A is a
protecting group, hydrogen, or hydroxyl.
[0366] As noted above, compounds in which R.sub.A is a protecting
group can be stably stored until it becomes desirable to associate
the compound with a Bone Active Portion. Similarly, as also noted
above, salts derived from compounds in which R.sub.A is hydrogen or
hydroxyl can be stably stored until it becomes desirable to
associate the compound with a Bone Active Portion. For purposes of
the present example, the compounds of the presently-disclosed
subject matter where R.sub.A is a protecting group, hydrogen, or
hydroxyl will be referred to as precursor compounds.
[0367] A precursor compound can be selected depending on the
terminal group of the at least one substitution, as will be
understood by those of ordinary skill in the art. For example, if
the terminal group is an amino or a hydroxyl, it can be desirable
to select a precursor compound including a carboxyl capable of
condensing with the amino or the hydroxyl. The reaction can be
conducted under amide- or ester-forming conditions, as are known to
those of ordinary skill in the art.
Exemplary Synthesis where R.sub.A is Derived from an Anti-Cancer
Agent
##STR00314##
[0369] In a 100 mL round-bottom flask was combined 500 mg (1.35
mmol) NBR-VI-232-1, 155 mg (1.35 mmol) glutaric anhydride, 590
.mu.l DIEA, and 40 mL DMF. A homogeneous solution formed and it was
stirred overnight at room temperature. Approximately 20 mL of 50%
aq. MeOH was then added to the reaction solution and it was allowed
to stir for 30 minutes more. The solvent was removed under vacuum
to give a sticky brown solid. When triturated in methanol,
off-white precipitate NBR-VI-262-1 was noted and collected via
suction filtration. The yield was about 500 mg, 83%.
##STR00315##
[0370] 286 mg (0.65 mmol) of NBR-VI-262-1, along with 174 mg (1.29
mmol) of HOBt, 571 mg (1.29 mmol) BOP, and 225 .mu.l DIEA, were
dissolved in 25 mL DMF. After 5 minutes, 250 mg (0.43 mmol) of
doxorubicin.HCl was added to the stirring solution. The reaction
flask was wrapped in foil and purged with N.sub.2. The flask was
sealed and the reaction left stirring overnight.
[0371] After about 21 hours the DMF was removed under vacuum to
give a red oil. The oil was taken up in EtOAc and triturated. A red
solid precipitated. The mixture was centrifuged and the supernatant
removed. The solid was washed with EtOAc twice more, each time
removing the solvent by centrifugation/decanting. The red product,
NBR-VI-269-1, was purified via reverse-phase column chromatography
(Mega Bond Elut column). Product NBR-VI-274-1 was obtained after
the chromatographic separation. The yield was about 120 mg,
28.8%.
Exemplary Synthesis where R.sub.A is Derived from an Anti-Cancer
Agent
##STR00316##
[0372] Exemplary Synthesis where R.sub.A is Derived from an
Antimicrobial Agent
##STR00317##
[0374] The compound wherein R.sub.A was derived from vancomycin was
prepared as follows. Vancomycin HCl (0.100 g, 67.3 mmol) was
treated with N,Ndiisopropylethylamine (43 mg, 336 .mu.mol),
BT-2-(peg2)-NH3Cl (37 mg, 101 mmol), DMSO (1 mL) and DMF (1 mL).
The solution was cooled to 10.degree. C. and to this solution was
added HATU (38 mg, 101 mmol). After stirring for 12 h, the reaction
was deemed complete by LC/MS. The reaction mixture was directly
purified by preparative HPLC to provide 25 mg of the desired
conjugate.
Targeting Bone
[0375] A Hydroxyapatite (HA) Binding Assay is used to determine
whether compounds have an affinity for bone. Compounds of the
presently-disclosed subject matter having a Bone Targeting Portion
(R.sub.T) are studied using the HA Binding Assay. A 10.sup.-3 M
solution of each analyte was made in 100% dimethylsulfoxide (DMSO).
A 100 fold dilution was then made to form a 10.sup.-5 M solution in
50 mM Tris-HCl buffer, pH 7.4, 1% DMSO. Tetracycline was used as a
reference analyte and approximately 50% was adsorbed to HA at the
concentration of 10.sup.-5 M. The HA slurry was 0.5 g/100 mL 50 mM
Tris-HCl buffer, 1% DMSO.
[0376] For each analyte, two samples were prepared. For one sample,
1 mL of 10.sup.-5 M analyte and 100 .mu.L 50 mM Tris-HCl buffer, 1%
DMSO was pipetted into a microcentrifuge tube. For the second
sample, 1 mL of 10.sup.-5 M analyte and 100 .mu.L of the HA slurry
was pipetted into a microcentrifuge tube. The samples were mixed
gently by inversion for 4 minutes and then centrifuged at 12,000 g
for 3 minutes to sediment the HA contained in those samples. The
supernatant was transferred to another microcentrifuge tube.
[0377] An electronic spectral scan (ultraviolet-visible) from
220-520 nm was obtained for each analyte using a Varian Cary 300
Bio Scan. The blank was 50 mM Tris-HCl buffer, 1% DMSO. The
wavelength of maximum absorbance (.lamda..sub.max) was determined,
and the extinction coefficient (.epsilon.) was calculated using the
Beer-Lambert Law.
[0378] The absorbance of the samples incubated with HA was measured
at .lamda..sub.max, and the molar concentration of the analyte was
then determined using the Beer-Lambert Law and the previously
calculated extinction coefficient. The fraction adsorbed to HA for
each sample was subsequently calculated. Binding to Hydroxyapatite
is expressed as a binding index for each compound tested, adjusted
such that tetracycline had a binding index of 100. Data are set
forth in the following Table.
TABLE-US-00007 Binding to Hydroxyapatite Compound Binding Index 17,
.beta.-estradiol -8 tetracycline 100 bone targeted steroidal
estrogenic agent 160 (compound of Formula 131) bone targeted
steroidal estrogenic agent 100 (compound of Formula 132) bone
targeted steroidal estrogenic agent 160 (compound of Formula 133)
bone targeted nitric oxide agent 130 (compound of Formula 138) bone
targeted androgen 140 (compound of Formula 33) bone targeted
carbonic anhydrase inhibitor 20 (compound of Formula 35) bone
targeted anti-cancer agent 60 (compound of Formula 146) bone
targeted antimicrobial 80 (compound of Formula 151)
[0379] Other compounds of the presently-disclosed subject matter,
including bone targeted nonsteroidal estrogenic agents (e.g.,
compound including a Bone Active Portion derived from genistein),
and other bone targeted androgens (e.g., compound including a Bone
Active Portion derived from DHEA), are tested and determined to
have an affinity for bone.
Affecting Bone
[0380] Animals. Six month old, bilaterally ovariectomized (OVX) or
sham operated Sprague-Dawley female rats (Harlan Laboratories,
Indianapolis, Ind.) were maintained at the University of Louisville
Research Resources Center at 22.degree. C. with a 12-h light/dark
cycle and ad libitum access to tap water and rodent chow (Purina
Laboratory Rodent Diet 5001). All animal procedures were approved
by the Institutional Animal Care and Use Committee, which is
certified by the American Association for Accreditation of
Laboratory Animal Care.
[0381] In vivo experiments were conducted in order to investigate
the efficacy of compounds of the presently-disclosed subject
matter. In all of the experiments, OVX and sham operated rats were
randomly divided into groups of 5-7 animals. In all of the studies
experimental groups included: i) Sham-operated control (euthanized
six weeks post surgery as a pretreatment control), ii) OVX control
(euthanized six weeks post surgery as a pretreatment control), iii)
Sham control receiving vehicle, iv) OVX control receiving vehicle,
v) OVX receiving 17-ethinyl estradiol (equimolar concentration with
a selected test compound), vi) OVX receiving alendronate (1.6
mg/kg), and vii) OVX receiving parathyroid hormone 1-34 (PTH) (80
.mu.g/kg). 17-ethinyl estradiol (17 EE) is a free estrogenic agent
known to be orally active, and can serve as an example of an
anti-catabolic agent. Alendronate (Alen) is a bisphosphonate that
is currently used to treat osteoporosis (e.g., Fosamax.RTM., Merck
& Co., Inc), and can serve as an example of an anti-catabolic
agent. Parathyroid hormone 1-34 (PTH) is an agent that can serve as
an example of an anabolic agent. All compounds and vehicle (1% DMSO
in corn oil) were administered three times per week orally by
gavage except for PTH which was administered via a subcutaneous
injection in a volume of 0.5 ml/kg body mass thrice per week.
Compound administration was initiated 6 weeks following surgery and
lasted for 6 weeks (18 doses total).
[0382] Compounds of the presently-disclosed subject matter were
orally administered to OVX rats at various doses. As indicated
above all BTE's were administered three times per week for 6
weeks.
[0383] Following 6 weeks of treatment, blood was obtained from each
animal via cardiac puncture following an overnight fast and animals
were subsequently euthanized via carbon dioxide asphyxiation. Blood
was centrifuged immediately and the obtained serum samples were
aliquoted and stored at -70.degree. C. prior to analysis. Uteri
were removed and fresh weights were obtained. Uterine masses were
normalized to body mass at the end of the experiment. The left and
right femora and left and right tibiae were subsequently collected
from each animal, cleaned of soft tissue, and stored in saline at
4.degree. C. prior to analysis.
[0384] Estrogenic Effect.
[0385] Quantitative Determination of Lipid Metabolism (Total
Cholesterol, HDL, and LDL) in Rat Serum. Total cholesterol,
high-density lipoprotein (HDL), and low-density lipoprotein (LDL)
were quantitatively determined in the serum of rats in order to
evaluate the extraosseous effects of estrogen treatment on lipid
metabolism. Commercially available kits from Wako Diagnostics
(Richmond, Va.) were used for the evaluations as recommended by the
manufacturer.
[0386] Uterine Mass and Body Weight. Ovariectomized (OVX) animals
generally exhibit an increase in body weight; however, upon
treatment with free estrogen, animals generally exhibit a decrease
in body weight to a normal body weight. As such, an assessment of
whole body estrogenic effect exhibited in the animals can be made
by measuring body weight of the animals following treatment.
Following OVX, animals exhibit a decrease in uterine mass, and a
subsequent increase in uterine mass upon treatment with free
estrogen. Uterine mass can be used to as a measure of estrogenic
effect occurring in a tissue outside the bone in response to
treatment with a test compound or composition. Generally, a lower
uterine mass can be associated with a decreased risk of adverse
side effects associated with the test compound or composition. A
ratio of uterine mass to body weight can be used to correct for any
increase in uterine size attributable to the relative size of the
individual animal. Body weight and/or uterine mass also serve as an
indirect measures of toxicity of a test compound.
[0387] Bone Density.
[0388] Femoral Density via Archimedes' Principle. Right femora were
submerged in distilled water and fully hydrated under a vacuum for
1 hr. Subsequently, the mass of each hydrated femur was obtained in
air and when submerged in water. Densities were determined using
Archimedes' Principle according to the following formula:
density=[mass of hydrated femur/(mass of hydrated femur--mass of
hydrated femur submerged in water)].times.density of distilled
water at a given temperature (See Keenan, et al. Comparison of bone
density measurement techniques: DXA and Archimedes' principle. J
Bone Miner Res 1997; 12:1903-7.). The obtained density measurements
were associated with whole bone.
[0389] Regional Femoral Density via Archimedes' Principle. In
addition to whole bone measurements, assessments of the densities
at different regions of the bone were made. The proximal and distal
ends of the femur are comprised primarily of cancellous/trabecular
bone, whereas the femoral diaphysis contains primarily cortical
bone. It is in these regions of bone that problems, for example, in
osteoporosis typically occur. Evaluation of the density of each of
these femoral regions can thereby increase the understanding of the
effect of a compound on each of these types of bone. The left
femora were separated into three regions (proximal left femur,
distal left femur, and left femoral diaphysis) using an Isomet Low
Speed Precision Sectioning Saw from Buehler Limited (Lake Bluff,
Ill.) with a diamond blade. Briefly, each femur was measured with a
Cen-Tech Digital Caliper and a cut was made from each end at 20% of
the length of the femur plus half the width of the blade.
Subsequently, the bone marrow was washed out of the femoral
diaphysis and the Archimedes density for each of the three femoral
regions was determined as described above.
[0390] Volume Fraction--Ex vivo micro-computed tomography (.mu.CT).
Volume fraction is a representative value for the amount of bone
that occupies a given volume or space. High resolution image data
were collected using a customized micro-CT system (ACTIS 150/225
system, BIR Inc., Lincolnshire, Ill.). The metaphysis of each right
tibia was scanned over a three millimeter range and
three-dimensional images were reconstructed. Data were subsequently
processed to reveal the volume fraction (BV/TV) occupied by
trabecular (cancellous) bone tissue, cortical bone tissue, and
whole bone tissue. The resulting data provides information
regarding the density of whole bone, cortical bone, and trabecular
bone.
[0391] Bone Strength.
[0392] Bone Mechanical Competence Indentation Test. After
sacrifice, the left tibiae were trimmed to expose the cancellous
bone of the proximal tibial metaphysis and an indentation test was
performed by advancing a flat-tipped cylindrical post (1.5 mm
diameter) axially into the cut surface to measure the compressive
strength of the cancellous bone structure.
[0393] Bone Formation and Turnover.
[0394] Rat Osteocalcin EIA. Osteocalcin is a hydroxyapatite-binding
protein that is synthesized by osteoblasts during bone formation.
Thereby, serum osteocalcin levels are commonly used as a
biochemical marker of bone formation. Rat serum osteocalcin levels
were measured using the Rat Osteocalcin EIA Kit from Biomedical
Technologies, Incorporated (Stoughton, Mass.) as recommended by the
manufacturer.
[0395] RatLaps.TM. ELISA for C-telopeptide Fragments of Collagen
Type I (CTX-I). Osteoclast mediated breakdown of collagen type I in
bone leads to the release of free and peptide bound fragments of
the collagen type I molecule. The fragment released from the
carboxy-terminal region of collagen type I is termed the
C-telopeptide fragment of collagen type I (CTX-I) and is commonly
used as a biochemical marker of bone resorption. Bone resorption
(CTX-I) was quantitatively assessed in rat serum using the
commercially available RatLaps.TM. ELISA KIT from Nordic Bioscience
Diagnostics A/S (Herlev, Denmark) as recommended by the
manufacturer.
[0396] Stimulation of Periosteal Bone Formation in a Mouse
Calvarial Injection Model. The anabolic (bone formation) of
compounds of the presently-disclosed subject matter are evaluated
in the mouse calvarial injection model. Briefly, 4-week-old ICR
Swiss mice are injected subcutaneously over the surface of the
calvariae with compounds of the presently-disclosed subject matter
at concentrations of 0, 1, 3, and 10 mg/kg/day for 5 days (twice a
day). Microtubule inhibitor TN-16 (5 mg/kg/day for 2 days, twice a
day) are used as a positive control. Mice are sacrificed two weeks
after the injections are completed. Dissected calvarial samples are
fixed in 10% phosphate-buffered formalin for 2 days, decalcified in
10% EDTA for 2 weeks and then embedded in paraffin. Histological
sections are cut and stained with H&E and orange G. New woven
bone formation (new bone area) is quantified by histomorphometry
using the OsteoMeasure system (OsteoMetrics Inc., Atlanta,
Ga.).
[0397] Anti-Cancer Effect: Colony formation assay. The anti-cancer
effect of compounds including a bone active portion (R.sub.A)
derived from an anti-cancer agent was tested. SUM1315 human breast
cancer cells (derived from a clinical metastatic nodule) were
seeded in 6 well plates at 5000 cells/well in 2 ml of medium. Cells
were allowed to adhere overnight prior to drug treatment. Compounds
of the presently-disclosed subject matter including a bone active
portion (R.sub.A) derived from an anti-cancer agent, or the free
anti-cancer agent, were added to the cells at concentrations of
0.05, 0.1, 0.5, 1, 5, and 10 .mu.M and cells were allowed to grow
at 37.degree. C. for 10 days. In addition, untreated and vehicle
(0.1% DMSO) treated wells were included as controls. After 10 days
of treatment, medium was removed from each well and cells were
subsequently washed with water. Cells were then fixed in 10%
formaldehyde for 10 minutes, washed with water and then stained
with 0.5% crystal violet for 5 minutes. Colonies of cells were
subsequently counted under the microscope in three separate frames.
Three independent experiments were performed and the results are an
average of the three experiments. Images were obtained from a
representative well for each concentration.
[0398] Anti-Cancer Effect: Cell Proliferation Assay. The
colorometric MTT assay is based on the cleavage of a yellow MTT
tetrazolium salt to form purple formazan crystals by the
mitochondria of metabolically active and viable cells. The formazan
crystals are insoluble in aqueous solution but can be dissolved in
acid and quantified using a spectrophotometer. The number of living
and viable cells in a sample directly correlates to the amount of
purple formazan crystals formed. SUM1315 human breast cancer cells
were seeded in triplicate in 96 well plates at 1000 cells/well and
were allowed to adhere overnight prior to drug treatment. Compounds
of the presently-disclosed subject matter including a bone active
portion (R.sub.A) derived from an anti-cancer agent, or the free
anti-cancer agent, were added to the cells at concentrations of
0.05, 0.1, 0.5, 1, 5, and 10 .mu.M and cells were allowed to grow
at 37.degree. C. for 1, 2, 3, 4, 5, or 6 days. In addition,
untreated and vehicle (0.1% DMSO) treated wells were included at
each time point as controls. Following treatment the MTT assay was
subsequently performed and the absorbance at 490 nm was recorded.
Three independent experiments were performed and the results were
averaged.
[0399] Antimicrobial Effect: Minimum Inhibitory Concentration (MIC)
of a Test Compound Against Isolates of Staphylococcus aureus. The
Minimum inhibitory concentration (MIC) of compounds of the
presently-disclosed subject matter including a bone active portion
(R.sub.A) derived from an antimicrobial agent, and free
antimicrobial agents, were determined by Ricerca Biosciences LLC
(Concord, Ohio) using the standard in vitro broth microdilution
assay (CLSI). Two isolates of S. aureus, one resistant to
Methicillin (ATCC 33591) and one nonresistant to Methicillin (ATCC
49230) were used for the evaluation. ATCC 49230 is a clinical
isolate from a patient with chronic osteomyelitis. Additionally, a
quality control isolate (ATCC 29213) was included in the analysis.
Briefly, twelve, serial, one-half dilutions of compounds of the
presently-disclosed subject matter including a bone active portion
(R.sub.A) derived from an antimicrobial agent, and free
antimicrobial agents, were prepared in 96-well plates in Mueller
Hinton Broth (MHB). The highest concentration of each compound used
was 64 .mu.g/mL. Bacterial suspensions were prepared and added to
each well at a concentration of approximately 5.times.10.sup.5
colony-forming-units per milliliter. The inoculated plates were
incubated for 16-20 h at 35.+-.1.degree. C. At the completion of
incubation, the wells of each plate were evaluated visually for the
presence of bacterial growth. All testing was completed in
duplicate. The MIC is the concentration of the compound at which
growth of the bacteria was not visible.
Bone Targeting Portion (R.sub.T)
[0400] Animals were treated with the following compounds having an
affinity for bone as assessed by HA-binding assay, including bone
targeting portions (R.sub.T), but lacking linking portions
(R.sub.L) and bone active portions (R.sub.A). Samples were
collected and studied as described above.
##STR00318##
[0401] With reference to FIG. 1, the body weights of the animals
were measured at regular time intervals and body weights were
plotted as a function of time. Body weight can serve as an indirect
measure of toxicity. The compounds of Formulas 161 (BTA-2) and 162
(BTA-3) are shown to have no effect on body weight. Turning now to
FIGS. 2 and 3 the uterine mass of each animal was measured and
expressed both independently, and as a ratio of uterine mass to
body weight. The compounds of Formulas 161 (BTA-2) and 162 (BTA-3)
are shown to have no effect on uterine mass, nor ratio of uterine
mass to body weight.
[0402] The right femora were used to assess bone density, as
described above. With reference to FIG. 4, whole bone density was
not significantly effected by compounds of Formulas 161 (BTA-2) or
162 (BTA-3). Regional bone density was also assessed, as described
above. With reference to FIGS. 5 and 6, regional bone density was
not significantly effected by compounds of Formulas 161 (BTA-2) or
162 (BTA-3).
Compound including Bone Active Portion (R.sub.A) Derived from a
Steroidal Estrogenic Agent
[0403] Animals were treated with a compounds including a bone
active portion (R.sub.A) derived from a steroidal estrogenic agent.
Compounds wherein the bone active portion was derived from
estradiol were selected as examples of compounds including a bone
active portion derived from a steroidal estrogenic agent. The
compounds of Formulas 123, 124, and 125 were orally administered to
animals at doses of 10, 100, or 1000 .mu.g/kg.
[0404] Body Weight and Uterine Mass.
[0405] With reference to FIG. 7, the body weights of the animals
were measured at regular time intervals and body weights were
plotted as a function of time. Body weight can serve as an indirect
measure of toxicity. With increasing doses of the compound of
Formula 131 (BTE2-pg2-D2), body weight is shown to decrease,
approaching the sham animal when the compound of Formulas 123
(BTE2-pg2-D2) is administered at the highest concentration.
[0406] Turning now to FIGS. 8 and 9 the uterine mass of each animal
was measured and expressed both independently, and as a ratio of
uterine mass to body weight. Animals treated with the compound of
Formula 131 (BTE2-pg2-D2) had lower uterine masses, and uterine
mass to body weight ratios as compared to the animals treated with
the free steroidal estrogenic agent, 17-ethinyl estradiol. This is
a surprising result given that the compound of Formula 131 includes
a bone active portion derived from the steroidal estrogenic agent,
estradiol. These results indicate that the compounds of the
presently-disclosed subject matter including a bone active portion
derived from a estrogenic agent do not act in the same manner as
free estrogenic agents.
[0407] Lipid Metabolism.
[0408] Total cholesterol, high-density lipoprotein (HDL), and
low-density lipoprotein (LDL) in collected serum samples are
quantitatively determined. The compound of Formula 131
(BTE2-pg2-D2) have a limited or no effect on lipid metabolism.
[0409] Bone Density.
[0410] Femoral Density and Regional Femoral Density.
[0411] The right femora were used to assess bone density, as
described above. With reference to FIG. 10, whole bone density was
affected by the compound of Formula 131 (BTE2-pg2-D2). Animals
receiving Formula 131 (BTE2-pg2-D2) were shown to have whole bone
densities in the same ranges as those animals receiving 17-ethinyl
estradiol (17 EE), Alendronate (Alen), or Parathyroid hormone 1-34
(PTH).
[0412] Regional bone density was also assessed, as described above,
with data presented in FIGS. 11, 12, and 13. With reference to
FIGS. 11 and 12, treatment with the compound of Formula 131
(BTE2-pg2-D2) affected regional bone density. Animals receiving the
compound Formula 131 (BTE2-pg2-D2) were shown to have increased
regional bone densities, as compared to the OVX animals.
[0413] Volume Fraction--Trabecular, Cortical, and Whole Bone.
[0414] Samples are collected as described above. Data revealing the
volume fraction (BV/TV) occupied by trabecular (cancellous) bone
tissue, cortical bone tissue, and whole bone tissue is obtained.
The compound of Formula 131 is found to increase bone density.
[0415] Bone Strength.
[0416] Mechanical Competence.
[0417] Samples are collected and an indentation test is performed
as described above.
[0418] The compound of Formula 131 is found to increase bone
strength.
[0419] Bone Formation and Turnover.
[0420] Osteocalcin.
[0421] With reference to FIG. 14, serum osteocalcin levels were
measured as described above. Measurements associated with
pretreatment sham, pretreatment OVX, and BTE2-pg-D2 at the 10
.mu.g/kg dose were not made. The animals treated with the compound
of Formula 131 (BTE2-pg2-D2) received either 100 or 1000 .mu.g/kg
doses. These data show that the compound of Formula 131
(BTE2-pg2-D2) stimulates osteocalcin, which indicates that the
compound stimulates of bone formation in a manner similar to
PTH.
[0422] Calvarial Injection Model.
[0423] Animals are administered control agents and the compound,
samples are collected, and data are obtained, as described above.
The compound of Formula 131 is found to affect new bone area.
[0424] Body Weight and Uterine Mass.
[0425] With reference to FIG. 15, the body weights of the animals
were measured at regular time intervals and body weights were
plotted as a function of time. Body weight can serve as an indirect
measure of toxicity. With increasing doses of the compound of
Formula 132 (BTE2-pg3-D2), body weight is shown to decrease,
approaching the sham animals when the compound of Formulas 124
(BTE2-pg3-D2) is administered at increasing concentrations.
[0426] Turning now to FIGS. 16 and 17 the uterine mass of each
animal was measured and expressed both independently, and as a
ratio of uterine mass to body weight. Animals treated with the
compound of Formula 132 (BTE2-pg3-D2) had lower uterine masses, and
uterine mass to body weight ratios as compared to the animals
treated with the free steroidal estrogenic agent, 17-ethinyl
estradiol. This is a surprising result given that the compound of
Formula 132 includes a bone active portion derived from the
steroidal estrogenic agent, estradiol. These results indicate that
the compounds of the presently-disclosed subject matter including a
bone active portion derived from a estrogenic agent do not act in
the same manner as free estrogenic agents.
[0427] Lipid Metabolism.
[0428] Total cholesterol, high-density lipoprotein (HDL), and
low-density lipoprotein (LDL) in collected serum samples are
quantitatively determined. The compound of Formula 132 has a
limited or no effect on lipid metabolism.
[0429] Bone Density.
[0430] Femoral Density and Regional Femoral Density.
[0431] The right femora were used to assess bone density, as
described above. With reference to FIG. 18, whole bone density was
affected by the compound of Formula 132 (BTE2-pg3-D2). Animals
receiving Formula 132 (BTE2-pg3-D2) were shown to have whole bone
densities in the same ranges as those animals receiving 17-ethinyl
estradiol (17 EE), Alendronate (Alen), or Parathyroid hormone 1-34
(PTH).
[0432] Regional bone density was also assessed, as described above,
with data presented in FIGS. 19, 20, and 21. Treatment with the
compound of Formula 132 (BTE2-pg3-D2) affected regional bone
density. Animals receiving the compound Formula 132 (BTE2-pg3-D2)
were shown to have regional bone densities in the same ranges as
those animals receiving 17-ethinyl estradiol (17 EE), Alendronate
(Alen), or Parathyroid hormone 1-34 (PTH).
[0433] Volume Fraction--Trabecular, Cortical, and Whole Bone.
[0434] Bone density was further assessed by measuring trabecular
volume fraction using ex vivo micro-computed tomography. With
reference to FIG. 22, animals receiving various doses of the
compound of Formula 132 (BTE2-pg3-D2) were shown to have trabecular
volume fractions in the same ranges as those animals receiving
17-ethinyl estradiol (17 EE), Alendronate (Alen), or Parathyroid
hormone 1-34 (PTH). These data indicate that the compound of
Formula 132 (BTE2-pg3-D2) maintains and/or stimulates trabecular
bone to about the same degree as alendronate, estradiol, and
PTH.
[0435] FIG. 23 includes three-dimensional images that were
reconstructed using high resolution image data collected using the
customized micro-CT system described above. These data indicate
that the compound of Formula 132 (BTE2-pg3-D2) affected an increase
in bone density.
[0436] Bone Strength.
[0437] Mechanical Competence.
[0438] Bone strength was assessed using the bone mechanical
competence indentation test, as described above. With reference to
FIG. 24, animals receiving the compound of Formula 132
(BTE2-pg3-D2) were shown to have a dose-responsive increase in bone
strength, relative to the OVX animals, with bone strength at the
higher doses in the same ranges as bone strength for animals
receiving alendronate or parathyroid hormone.
[0439] Bone Formation and Turnover.
[0440] Calvarial Injection Model.
[0441] Animals are administered control agents and the compound,
samples are collected, and data are obtained, as described above.
The compound of Formula 132 is found to increase new bone area.
[0442] Body Weight and Uterine Mass.
[0443] With reference to FIG. 25, the body weights of the animals
were measured at regular time intervals and body weights were
plotted as a function of time. Body weight can serve as an indirect
measure of toxicity. Animals receiving the compound of Formula 133
(BTE2-pg2-D3) tend to have lower body weights as compared to the
animals receiving alendronate or parathyroid hormone.
[0444] Turning now to FIGS. 26 and 27 the uterine mass of each
animal was measured and expressed both independently, and as a
ratio of uterine mass to body weight. Animals treated with the
compound of Formula 133 (BTE2-pg2-D3) had lower uterine masses, and
uterine mass to body weight ratios as compared to the animals
treated with the free steroidal estrogenic agent, 17-ethinyl
estradiol. This is a surprising result given that the compound of
Formula 133 includes a bone active portion derived from the
steroidal estrogenic agent, estradiol. These results indicate that
the compounds of the presently-disclosed subject matter including a
bone active portion derived from a estrogenic agent do not act in
the same manner as free estrogenic agents.
[0445] Lipid Metabolism.
[0446] Total cholesterol, high-density lipoprotein (HDL), and
low-density lipoprotein (LDL) in collected serum samples are
quantitatively determined. The compound of Formula 133 has a
limited or no effect on lipid metabolism.
[0447] Bone Density.
[0448] Femoral Density and Regional Femoral Density.
[0449] Bone density was also assessed, as described above, with
data presented in FIGS. 28, 29, 30, and 31. The compound of Formula
133 (BTE2-pg2-D3) affected regional bone density. Animals receiving
the compound Formula 133 (BTE2-pg2-D3) were shown to have increased
regional bone densities, as compared to the OVX animals. At certain
doses, animals receiving the compound Formula 133 were shown to
have increased regional bone densities, as compared to the Sham
animals.
[0450] Volume Fraction--Trabecular, Cortical, and Whole Bone.
[0451] Bone density was further assessed by measuring trabecular
volume fraction using ex vivo micro-computed tomography. FIG. 32
includes the trabecular volume fraction data for control animals
and animals receiving various doses of the compound of Formula 133
(BTE2-pg2-D3). These data indicate that bone density is increased,
as compared to OVX animals, in animals receiving the compound of
Formula 133.
[0452] With reference to FIG. 33, three-dimensional images were
reconstructed using high resolution image data collected using the
customized micro-CT system described above. These data indicate
that the compound of Formula 133 (BTE2-pg2-D3) affected an increase
in bone density.
[0453] Bone Strength.
[0454] Mechanical Competence.
[0455] Samples are collected and an indentation test is performed
as described above.
[0456] The compound of Formula 133 is found to increase bone
strength.
[0457] Bone Formation and Turnover.
[0458] Collagen Type I.
[0459] Bone resorption or osteoclast-mediated breakdown of collagen
type I in bone was assessed by measuring the C-telopeptide fragment
of collagen type I (CTX-I), as described above. With reference to
FIG. 34, it is shown that treatment with 1000 .mu.g/kg doses of the
compound of Formula 133 (BTE2-pg2-D3) do not inhibit production of
CTX-I, indicating that bone resorption is not inhibited and that
the compound acts in a similar manner to known anabolic agent,
parathyroid hormone 1-34 (PTH). In contrast, known anti-catabolic
agents, 17-ethinyl estradiol (17-EE) and alendronate (Alen),
descrease the production of CTX-1 to sham levels.
[0460] Calvarial Injection Model.
[0461] Animals are administered control agents and the compound,
samples are collected, and data are obtained, as described above.
The compound of Formula 133 is found to increase new bone area.
[0462] Compound including Bone Active Portion (R.sub.A) Derived
from an Androgen.
[0463] Animals were treated with a compounds including a bone
active portion (R.sub.A) derived from an androgen. Compounds
wherein the bone active portion was derived from testosterone were
selected as examples of compounds including a bone active portion
derived from an androgen. The compound of Formula 33
(BT-Testosterone) was orally administered to animals at a dose of
3000 .mu.g/kg.
[0464] Body Weight and Uterine Mass.
[0465] With reference to FIG. 35, the body weights of the animals
were measured at regular time intervals and body weights were
plotted as a function of time. Body weight can serve as an indirect
measure of toxicity. Animals receiving the compound of Formula 33
(BT-Testosterone) tend to have lower body weights as compared to
the OVX animals.
[0466] Turning now to FIGS. 36 and 37 the uterine mass of each
animal was measured and expressed both independently, and as a
ratio of uterine mass to body weight. Animals treated with the
compound of Formula 33 (BT-Testosterone) had lower uterine masses,
and uterine mass to body weight ratios as compared to the animals
treated with the free steroidal estrogenic agent, 17-ethinyl
estradiol.
[0467] Lipid Metabolism.
[0468] Total cholesterol, high-density lipoprotein (HDL), and
low-density lipoprotein (LDL) in collected serum samples are
quantitatively determined. The compound of Formula 33 has a limited
or no effect on lipid metabolism.
[0469] Bone Density.
[0470] Femoral Density and Regional Femoral Density.
[0471] The right femora were used to assess bone density, as
described above. With reference to FIG. 38, whole bone density was
affected by the compound of Formula 33 (BT-Testosterone). Animals
receiving Formula 33 (BT-Testosterone) were shown to have
dose-responsive whole bone densities that were higher than that of
the OVX animals, and in the same ranges as those animals receiving
17-ethinyl estradiol (17 EE), Alendronate (Alen), or Parathyroid
hormone 1-34 (PTH).
[0472] Regional bone density was also assessed, as described above,
with data presented in FIGS. 39, and 40. The compound of Formula 33
(BT-Testosterone) affected regional bone density. Animals receiving
the compound Formula 33 (BT-Testosterone) were shown to have
regional bone densities that were higher than that of the OVX
animals, and in the same ranges as those animals receiving
17-ethinyl estradiol (17 EE), Alendronate (Alen), or Parathyroid
hormone 1-34 (PTH).
[0473] Volume Fraction--Trabecular, Cortical, and Whole Bone.
[0474] Samples are collected as described above. Data revealing the
volume fraction (BV/TV) occupied by trabecular (cancellous) bone
tissue, cortical bone tissue, and whole bone tissue is obtained.
The compound of Formula 33 is found to increase bone density.
[0475] Bone Strength.
[0476] Mechanical Competence.
[0477] Samples are collected and an indentation test is performed
as described above. The compound of Formula 33 is found to increase
bone strength.
[0478] Bone Formation and Turnover.
[0479] Calvarial Injection Model.
[0480] Animals are administered control agents and the compound,
samples are collected, and data are obtained, as described above.
The compound of Formula 33 is found to increase new bone area.
Compound including Bone Active Portion (R.sub.A) Derived from a
Non-Steroidal Estrogenic Agent
[0481] Animals are administered control agents and the compound of
Formula 136. A compound wherein the bone active portion was derived
from genistein was selected as an example of compounds including a
bone active portion derived from a non-steroidal estrogenic agent.
Samples are collected and studied as described above.
[0482] Uterine Mass.
[0483] Uterine mass is obtained. The compound of Formula 136 has a
limited or no effect on uterine mass.
[0484] Lipid Metabolism.
[0485] Total cholesterol, high-density lipoprotein (HDL), and
low-density lipoprotein (LDL) in collected serum samples are
quantitatively determined. The compound of Formula 136 has a
limited or no effect on lipid metabolism.
[0486] Bone Density.
[0487] Femoral Density and Regional Femoral Density.
[0488] Samples are collected as described above. Whole bone density
and regional bone density are assessed. The compound of Formula 136
is found to increase bone density.
[0489] Volume Fraction--Trabecular, Cortical, and Whole Bone.
[0490] Samples are collected as described above. Data revealing the
volume fraction (BV/TV) occupied by trabecular (cancellous) bone
tissue, cortical bone tissue, and whole bone tissue is obtained.
The compound of Formula 136 is found to increase bone density.
[0491] Bone Strength.
[0492] Mechanical Competence.
[0493] Samples are collected and an indentation test is performed
as described above. The compound of Formula 136 is found to
increase bone strength.
[0494] Bone Formation and Turnover.
[0495] Calvarial Injection Model.
[0496] Animals are administered control agents and the compound,
samples are collected, and data are obtained, as described above.
The compound of Formula 136 is found to increase new bone area.
[0497] Compound Including Bone Active Portion (R.sub.A) Derived
from a Nitric Oxide Agent
[0498] Animals are administered control agents and the compound of
Formula 138. A compound wherein the bone active portion was derived
from alkoxy-NO.sub.2 was selected as an example of compounds
including a bone active portion derived from a nitric oxide agent.
Samples are collected and studied as described above.
[0499] Uterine Mass.
[0500] Uterine mass is obtained. The compound of Formula 138 has a
limited or no effect on uterine mass.
[0501] Lipid Metabolism.
[0502] Total cholesterol, high-density lipoprotein (HDL), and
low-density lipoprotein (LDL) in collected serum samples are
quantitatively determined. The compound of Formula 138 has a
limited or no effect on lipid metabolism.
[0503] Bone Density.
[0504] Femoral Density and Regional Femoral Density.
[0505] Samples are collected as described above. Whole bone density
and regional bone density are assessed. The compound of Formula 138
is found to increase bone density.
[0506] Volume Fraction--Trabecular, Cortical, and Whole Bone.
[0507] Samples are collected as described above. Data revealing the
volume fraction (BV/TV) occupied by trabecular (cancellous) bone
tissue, cortical bone tissue, and whole bone tissue is obtained.
The compound of Formula 138 is found to increase bone density.
[0508] Bone Strength.
[0509] Mechanical Competence.
[0510] Samples are collected and an indentation test is performed
as described above. The compound of Formula 138 is found to
increase bone strength.
[0511] Bone Formation and Turnover.
[0512] Calvarial Injection Model.
[0513] Animals are administered control agents and the compound,
samples are collected, and data are obtained, as described above.
The compound of Formula 138 is found to increase new bone area.
Compound Including Bone Active Portion (R.sub.A) Derived from a
Carbonic Anhydrase Inhibitor
[0514] Animals are administered control agents and the compound of
Formula 35. A compound wherein the bone active portion was derived
from 2-amino-1,3,4-thiadiazole-5-sulfonamide was selected as an
example of compounds including a bone active portion derived from a
carbonic anhydrase inhibitor. Samples are collected and studied as
described above.
[0515] Uterine Mass.
[0516] Uterine mass is obtained. The compound of Formula 35 has a
limited or no effect on uterine mass.
[0517] Lipid Metabolism.
[0518] Total cholesterol, high-density lipoprotein (HDL), and
low-density lipoprotein (LDL) in collected serum samples are
quantitatively determined. The compound of Formula 35 has a limited
or no effect on lipid metabolism.
[0519] Bone Density.
[0520] Femoral Density and Regional Femoral Density.
[0521] Samples are collected as described above. Whole bone density
and regional bone density are assessed. The compound of Formula 35
is found to increase bone density.
[0522] Volume Fraction--Trabecular, Cortical, and Whole Bone.
[0523] Samples are collected as described above. Data revealing the
volume fraction (BV/TV) occupied by trabecular (cancellous) bone
tissue, cortical bone tissue, and whole bone tissue is obtained.
The compound of Formula 35 is found to increase bone density.
[0524] Bone Strength.
[0525] Mechanical Competence.
[0526] Samples are collected and an indentation test is performed
as described above. The compound of Formula 35 is found to increase
bone strength.
[0527] Bone Formation and Turnover.
[0528] Calvarial Injection Model.
[0529] Animals are administered control agents and the compound,
samples are collected, and data are obtained, as described above.
The compound of Formula 35 is found to increase new bone area.
[0530] Compound including Bone Active Portion (R.sub.A) Derived
from an Anti-Cancer Agent
[0531] Cells were treated with a compounds including a bone active
portion (R.sub.A) derived from an anti-cancer agent, as described
above. Compounds wherein the bone active portion was derived from
doxorubicin were selected as examples of compounds including a bone
active portion derived from an anti-cancer agent. Cells were
treated with the compound of Formula 146 (BT2-pg2-doxorubicin
(Nbr-VI)), and free doxorubicin, at concentrations of 0.05, 0.1,
0.5, 1, 5, and 10 .mu.M.
[0532] With reference to FIGS. 41A and 41B, there is a
dose-responsive decrease in colony formation in cells treated with
the compound of Formula 146 (BT2-pg2-doxorubicin (Nbr-VI)), showing
that the compound is effective against cancer cells.
[0533] FIG. 42 includes the results of treatment with doxorubicin
on the cells. With reference to FIGS. 43-49, there is a
dose-responsive decrease in cell proliferation in cells treated
with the compound of Formula 146 (BT2-pg2-doxorubicin (Nbr-VI)),
showing that the compound is effective against cancer cells, and
that the efficacy is similar to that of doxorubicin.
Compound Including Bone Active Portion (R.sub.A) Derived from an
Antimicrobial Agent
[0534] Isolates of S. Aureus were treated with a compounds
including a bone active portion (R.sub.A) derived from an
antimicrobial agent, as described above. Compounds wherein the bone
active portion was derived from vancomycin were selected as
examples of compounds including a bone active portion derived from
an antimicrobial agent. Cells were treated with the compound of
Formula 151 (BT2-pg2-vancomycin), and free vancomycin. The minimum
inhibitory concentrations (MIC) of the compound of Formula 151
(BT2-pg2-vancomycin) and standard (.mu.g/mL) against strains of
Staphylococcus aureus are set forth in the following table. These
results indicate that that the compound of Formula 151
(BT2-pg2-vancomycin) is as effective as vancomycin against the
tested strains of S. aureus.
TABLE-US-00008 ATCC# Compound 49230.sup.a 33591.sup.b 29213.sup.c
BT2-pg2-vancomycin 1 2 1 Vancomycin 1 1 1 .sup.aWild-type isolated
from bone infection; .sup.bMRSA; .sup.cQC strain: Acceptable
vancomycin MIC range: 0.5-2 .mu.g/mL
[0535] Throughout this document, various references are mentioned.
All such references are incorporated herein by reference, including
the references set forth in the following list: [0536] Keenan M J,
Hegsted M, Jones K L, Delany J P, Kime J C, Melancon L E, Tulley R
T, Hong K D. Comparison of bone density measurement techniques: DXA
and Archimedes' principle. J Bone Miner Res 1997; 12:1903-7. [0537]
Mundy G R, Garrett R, Harris S E, Chan J, Chen D, Rossini G, Boyce
B, Zhao M, and Gutierrez G (1999) Stimulation of bone formation in
vitro and in rodents by statins. Science 286:1946-1949. [0538]
Garrett I R, Chen D, Gutierrez G, Rossini G, Zhao M, Escobedo A,
Kim K B, Hu S, Crews C M, and Mundy G R (2003) Selective inhibitors
of the osteoblast proteasome stimulate bone formation in vivo and
in vitro. J Clin Invest 111:1771-1782. [0539] Seibel M J.
Biochemical Markers of Bone Turnover. Clin Biochem Rev. 2005;
26:97-122. [0540] Delmas P D, et al. The Use of Biochemical Markers
of Bone Turnover in Osteoporosis. Osteoporosis Int. suppl. 6S2-17
(2000). [0541] Riggs B L, and Parfitt A M, "Drugs Used to Treat
Osteoporosis: The Critical Need for a Uniform Nomenclature Based on
Their Action on Bone Remodeling," J. Bone and Mineral Res. 20:2
(2005). [0542] U.S. patent application Ser. Nos. 11/022,024;
11/021,661; and 11/674,753, and PCT Patent Publication No.
WO/0066613.
* * * * *