U.S. patent application number 14/345661 was filed with the patent office on 2014-09-04 for bone morphogenetic protein pathway activation, compositions for ossification, and methods related thereto.
This patent application is currently assigned to EMORY UNIVERSITY. The applicant listed for this patent is Scott D. Boden, Sreedhara Sangadala. Invention is credited to Scott D. Boden, Sreedhara Sangadala.
Application Number | 20140248372 14/345661 |
Document ID | / |
Family ID | 47914783 |
Filed Date | 2014-09-04 |
United States Patent
Application |
20140248372 |
Kind Code |
A1 |
Boden; Scott D. ; et
al. |
September 4, 2014 |
BONE MORPHOGENETIC PROTEIN PATHWAY ACTIVATION, COMPOSITIONS FOR
OSSIFICATION, AND METHODS RELATED THERETO
Abstract
The disclosure relates to compounds and compositions for bone
formation, fracture treatment, bone grafting, bone fusion,
cartilage maintenance and repair and methods related thereto. In
certain embodiments, the disclosure relates to compositions
comprising one or more compound(s) disclosed herein, such as
clotrimazole, honokiol, magnolol, tacrolimus, pimecrolimus,
sirolimus, everolimus, temsirolimus, spironolactone, fluticasone,
fluticasone propionate, fluticasone furoate, linezolid,
telmisartan, chlorambucil, retinol, isotretinoin, acitretin,
etretinate, retinoic acid (tretinoin), teniposide, mitomycin C,
cytarabine, decitabine, vinblastine, vincristine, vindesine,
vinorelbine, valrubicin, doxorubicin, daunorubicin, epirubicin,
idarubicin, mitoxantrone, pixantrone, plicamycin, pazopanib,
topotecan, camptothecin, irinotecan, sunitinib, derivatives, or
salt thereof, for use in bone growth processes. In a typical
embodiment, a bone graft composition is implanted in a subject at a
site of desired bone growth or enhancement.
Inventors: |
Boden; Scott D.; (Atlanta,
GA) ; Sangadala; Sreedhara; (Dallas, GA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Boden; Scott D.
Sangadala; Sreedhara |
Atlanta
Dallas |
GA
GA |
US
US |
|
|
Assignee: |
EMORY UNIVERSITY
Atlanta
GA
|
Family ID: |
47914783 |
Appl. No.: |
14/345661 |
Filed: |
September 17, 2012 |
PCT Filed: |
September 17, 2012 |
PCT NO: |
PCT/US12/55722 |
371 Date: |
March 19, 2014 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
61536168 |
Sep 19, 2011 |
|
|
|
61657099 |
Jun 8, 2012 |
|
|
|
61669199 |
Jul 9, 2012 |
|
|
|
Current U.S.
Class: |
424/602 ;
514/16.7; 514/291; 514/8.8; 540/456 |
Current CPC
Class: |
A61K 33/42 20130101;
A61L 27/54 20130101; A61K 31/506 20130101; A61K 31/7048 20130101;
A61L 27/24 20130101; A61K 31/475 20130101; A61L 27/46 20130101;
A61K 31/4174 20130101; A61K 31/232 20130101; A61K 31/05 20130101;
A61K 38/1875 20130101; A61L 27/52 20130101; A61K 31/196 20130101;
A61K 31/704 20130101; A61L 2300/414 20130101; A61K 31/4745
20130101; A61K 31/407 20130101; A61K 31/07 20130101; A61K 31/4184
20130101; A61K 31/136 20130101; A61K 31/7068 20130101; C08L 89/06
20130101; A61K 2300/00 20130101; A61K 31/585 20130101; A61K 33/42
20130101; A61K 38/39 20130101; A61L 27/46 20130101; A61K 31/404
20130101; A61L 27/12 20130101; A61L 2430/02 20130101; A61K 31/203
20130101; A61K 31/56 20130101; A61L 2300/45 20130101; A61K 31/5377
20130101; A61K 31/436 20130101; A61P 19/00 20180101 |
Class at
Publication: |
424/602 ;
540/456; 514/291; 514/8.8; 514/16.7 |
International
Class: |
A61K 31/436 20060101
A61K031/436; A61K 33/42 20060101 A61K033/42; A61K 38/18 20060101
A61K038/18 |
Claims
1. A bone graft composition comprising clotrimazole, honokiol,
magnolol, tacrolimus, pimecrolimus, sirolimus, everolimus,
temsirolimus, spironolactone, fluticasone, fluticasone propionate,
fluticasone furoate, linezolid, telmisartan, chlorambucil, retinol,
isotretinoin, acitretin, etretinate, retinoic acid (tretinoin),
teniposide, mitomycin C, cytarabine, decitabine, vinblastine,
vincristine, vindesine, vinorelbine, valrubicin, doxorubicin,
daunorubicin, epirubicin, idarubicin, mitoxantrone, pixantrone,
plicamycin, pazopanib, topotecan, camptothecin, irinotecan,
sunitinib, derivatives, or salts thereof.
2. The bone graft composition of claim 1, further comprising a
growth factor.
3. The bone graft composition of claim 2, wherein the growth factor
is a bone morphogenetic protein.
4. The bone graft composition of claim 3, wherein the bone
morphogenetic protein is BMP-2, BMP-5, BMP-6, BMP-7, or BMP-9.
5. The bone graft composition of claim 1, further comprising
calcium phosphates.
6. The bone graft composition of claim 5, wherein said calcium
phosphates are hydroxyapatite and tricalcium phosphate.
7. The bone graft composition of claim 1, further comprising a
collagen or hydrogel matrix.
8. A kit comprising a clotrimazole, honokiol, magnolol, tacrolimus,
pimecrolimus, sirolimus, everolimus, temsirolimus, spironolactone,
fluticasone, fluticasone propionate, fluticasone furoate,
linezolid, telmisartan, chlorambucil, retinol, isotretinoin,
acitretin, etretinate, retinoic acid (tretinoin), teniposide,
mitomycin C, cytarabine, decitabine, vinblastine, vincristine,
vindesine, vinorelbine, valrubicin, doxorubicin, daunorubicin,
epirubicin, idarubicin, mitoxantrone, pixantrone, plicamycin,
pazopanib, topotecan, camptothecin, irinotecan, sunitinib,
derivatives, or salt thereof and a graft composition.
9. The kit of claim 8, further comprising a bone morphogenetic
protein.
10-25. (canceled)
26. A bone graft comprising more than 10, 15, 20, 25, 30, 35 mM of
a compound disclosed herein per about 100 mm.sup.3, or about 150
mm.sup.3, or 50 to 200 mm.sup.3, or about 100 to 200 mm.sup.3 of
bone graft volume.
27. The bone graft of claim 26, wherein the compound is tacrolimus,
pimecrolimus, sirolimus, everolimus, or temsirolimus
Description
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This application claims priority to U.S. Provisional
Application No. 61/536,168 filed on Sep. 19, 2011 and U.S.
Provisional Application No. 61/657,099 filed on Jun. 8, 2012, and
U.S. Provisional Application No. 61/669,199 filed on Jul. 9, 2012,
all hereby incorporated by reference in their entirety.
FIELD
[0002] This disclosure relates to compounds and compositions for
bone formation, fracture treatment, bone grafting, bone fusion,
cartilage maintenance and repair, and methods related thereto. In
certain embodiments, the disclosure relates to bone graft
compositions comprising one or more compound(s) disclosed herein
such as clotrimazole, honokiol, magnolol, tacrolimus, pimecrolimus,
sirolimus, everolimus, temsirolimus, spironolactone, fluticasone,
fluticasone propionate, fluticasone furoate, linezolid,
telmisartan, chlorambucil, retinol, isotretinoin, acitretin,
etretinate, retinoic acid (tretinoin), teniposide, mitomycin C,
cytarabine, decitabine, vinblastine, vincristine, vindesine,
vinorelbine, valrubicin, doxorubicin, daunorubicin, epirubicin,
idarubicin, mitoxantrone, pixantrone, plicamycin, pazopanib,
topotecan, camptothecin, irinotecan, sunitinib, derivatives, or
salts thereof. In a typical embodiment, a bone graft composition is
implanted in a subject at a site of desired bone growth or
enhancement.
BACKGROUND
[0003] Bone grafting is typically performed for spinal fusions,
after cancerous bone removal, and in certain operations, e.g.,
plastic surgery. The iliac crest is often used as a donor site for
autologous grafts. Complications collecting bone from the iliac
crest include pain, nerve damage, hematoma and wound complications,
avulsion of the anterior superior iliac spine (ASIS), hematoma,
herniation of the abdominal cavity contents, and cosmetic
deformity. Thus, it is desirable to develop materials and methods
of forming bone that do not require harvesting bone from remote
sites of the patient.
[0004] Synthetic bone grafts typically include a matrix that holds
minerals and other salts. Natural bone has an intracellular matrix
mainly composed of type I collagen, and some synthetic bone grafts
include a collagen matrix. Synthetic bone grafts typically contain
bone growth factors such as bone morphogenetic proteins (BMPs)
because of their ability to induce ossification in the matrix
material. Recombinant human BMP-2 has been approved by the FDA in
synthetic bone grafts such as INFUSE.TM.. INFUSE.TM. is approved
for open tibial shaft fractures, lumbar interbody fusion, and sinus
and alveolar ridge augmentations. However, the high cost and need
for high concentrations of BMP-2 for treatment creates a barrier
for routine clinical use. Thus, there is a need to identify
additional compositions that can substitute or complement the use
of BMPs in treating bone-related conditions.
[0005] Conflicting reports provide that sirolimus and everolimus
have a bearing on osteoblast proliferation and differentiation or
decreasing osteoclast-mediated bone resorption. See Kalantar-Zedah
et al., Curr Opin Nephrol Hypertens., 2012, 21(4):389-403, Lee et
al., Stem Cells Dev., 2010, 19(4):557-68, WO2009017269,
WO2005/005434 (U.S. application Ser. No. 12/398,225), U.S. Pat.
Nos. 5,258,389 and 6,015,815. References cited herein are not an
admission of prior art.
SUMMARY
[0006] This disclosure relates to compounds and compositions for
ossification and methods related thereto. In certain embodiments,
it is an object of the disclosure to provide certain compounds,
compositions, and methods of using these compounds to improve bone
formation, fracture treatment, bone grafting, bone fusion,
cartilage maintenance and repair in a subject. In a typical
embodiment, the bone graft composition comprises a compound
disclosed herein or derivatives that modulates BMP and/or BMPR
interactions with their natural target proteins. In specific
embodiments, the disclosure relates to compounds, such as
1-triarylmethyl-1H-imidazole derivatives, or salts thereof, such as
clotrimazole; biphenyl-diol derivatives, or salts thereof such as
honokiol and magnolol; macrolide lactone derivatives, or salts
thereof such as tacrolimus, pimecrolimus, sirolimus, everolimus,
temsirolimus; steroid derivatives, or salts thereof such as
spironolactone;
3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenant-
hrene-17-carbothioic acid derivatives, or salts thereof such as
fluticasone, fluticasone propionate, and fluticasone furoate;
3-(4-morpholinophenyl)oxazolidin-2-one derivatives, or salts
thereof such as linezolid; 1H,3'H-2,5'-bibenzo[d]imidazole
derivatives or salts thereof such as telmisartan; rentinol
derivatives, or salts thereof such as tretinoin, alitretinoin,
isotretinoin, retinol, etretinate, acitretin;
4-(4-(dialkylamino)phenyl)butanoic acid derivatives, or salts
thereof such as chlorambucil; podophyllotoxin derivatives such as
teniposide; aziridine derivatives such as mitomycin C; nucleoside
derivatives such as cytarabine, decitabine; vinca alkaloid
derivatives such as vinorelbine, vinblastine, vincristine, and
vindesine; anthracycline doxorubicin derivatives such as
doxorubicin, valrubicin, daunorubicin, epirubicin, idarubicin;
anthraquinone derivatives such as mitoxantrone and pixantrone;
plicamycin or derivatives; pazopanib or derivatives; camptothecin
or derivatives such as topotecan, irinotecan, 9-aminocamptothecin;
sunitinib or derivatives; and compositions including such
compounds, as well as their methods of use.
[0007] Examples of additional contemplated compounds include
1-[(2-chlorophenyl)diphenylmethyl]-1H-imidazole, derivatives, or
salts thereof, such as clotrimazole;
3',5-di-2-propen-1-yl-[1,1'-biphenyl]-2,4'-diol, derivatives, or
salts thereof such as honokiol;
5,6,8,11,12,13,14,15,16,17,18,19,24,25,26,26a-hexadecahydro-5,19-dihydrox-
y-3-[(1E)-2-[4-hydroxy-3-methoxycyclohexyl]-1-methylethenyl]-14,16-dimetho-
xy-4,10,12,18-tetramethyl-8-(2-propen-1-yl)-15,19-epoxy-3H-pyrido[2,1-c][1-
,4]oxaazacyclotricosine-1,7,20,21(4H,23H)-tetrone, derivatives, or
salts thereof such as tacrolimus;
6,9-difluoro-11-hydroxy-16-methyl-3-oxo-17-(1-oxopropoxy)-androsta-1,4-di-
ene-17-carbothioic acid S-(fluoromethyl) ester, derivatives, or
salts thereof such as fluticasone propionate, derivatives, or salts
thereof;
N-[[(5S)-3-[3-fluoro-4-(4-morpholinyl)phenyl]-2-oxo-5-oxazolidinyl]methyl-
]-acetamide, derivatives, or salts thereof such as linezolid; and,
4'-((1,7'-dimethyl-2'-propyl-1H,3'H-[2,5'-bibenzo[d]imidazol]-3'-yl)methy-
l)-[1,1'-biphenyl]-2-carboxylic acid, derivatives, or salts thereof
such as telmisartan; and
9,10,12,13,14,21,22,23,24,25,26,27,32,33,34,34a-Hexadecahydro-9,27-dihydr-
oxy-3-[2-[4-hydroxy-3-methoxycyclohexyl]-1-methylethyl]-10,21-dimethoxy-6,-
8,12,14,20,26-hexamethyl-23,27-epoxy-3H-pyrido[2,1-c][1,4]oxaazacyclohentr-
iacontine-1,5,11,28,29(4H,6H,31H)-pentone, derivatives, or salts
thereof such as sirolimus, everolimus, and temsirolimus.
[0008] Other contemplated compounds include those from the
following list: capsaicin--chemical name
N-(4-hydroxy-3-methoxybenzyl)-8-methylnon-6-enamide;
dihydrocapsaicin; nordihydrocapsaicin; homodihydrocapsaicin;
homocapsaicin; nonivamide; diphenylcyclopropenone--chemical name
2,3-diphenylcycloprop-2-enone; estradiol--chemical name
(8R,9S,13S,14S,17S)-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyc-
lopenta[a]phenanthrene-3,17-diol; ketoconazole--chemical name
1-(4-(4-(((2R,4S)-2-((1H-imidazol-1-yl)methyl)-2-(2,4-dichlorophenyl)-1,3-
-dioxolan-4-yl)methoxy)phenyl)piperazin-1-yl)ethanone;
ethynylestradiol--chemical name
(8R,9S,13S,14S,17R)-17-ethynyl-13-methyl-7,8,9,11,12,13,14,15,16,17-decah-
ydro-6H-cyclopenta[a]phenanthrene-3,17-diol; progesterone--chemical
name
(8S,9S,10R,13S,14S,17S)-17-acetyl-10,13-dimethyl-6,7,8,9,10,11,12,13,14,1-
5,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-3(2H)-one;
salbutamol--chemical name
4-(2-(tert-butylamino)-1-hydroxyethyl)-2-(hydroxymethyl)phenol;
zardaverine--chemical name
6-(4-(difluoromethoxy)-3-methoxyphenyl)pyridazin-3-ol;
riluzole--chemical name
6-(trifluoromethoxy)benzo[d]thiazol-2-amine;
9-chloro-2-(furan-2-yl)-[1,2,4]triazolo[1,5-c]quinazolin-5-amine;
prazosin--chemical name
(4-(4-amino-6,7-dimethoxyquinazolin-2-yl)piperazin-1-yl)(furan-2-yl)metha-
none hydrochloride hydrate; urapidil--chemical name
6-((3-(4-(2-methoxyphenyl)piperazin-1-yl)propyl)amino)-1,3-dimethylpyrimi-
dine-2,4(1H,3H)-dione hydrochloride; naftopidil; carmofur--chemical
name
5-fluoro-N-hexyl-2,4-dioxo-3,4-dihydropyrimidine-1(2H)-carboxamide;
5-fluoro-N-alkyl-2,4-dioxo-3,4-dihydropyrimidine-1(2H)-carboxamide;
fluorouracil; itraconazole--chemical name
4-(4-(4-(4-((2-((1H-1,2,4-triazol-1-yl)methyl)-2-(2,4-dichlorophenyl)-1,3-
-dioxolan-4-yl)methoxy)phenyl)piperazin-1-yl)phenyl)-1-(sec-butyl)-1H-1,2,-
4-triazol-5(4H)-one; posaconazole; trimebutine--chemical name
2-(dimethylamino)-2-phenylbutyl 3,4,5-trimethoxybenzoate;
piceid--chemical name
(2S,3R,4S,5S,6R)-2-(3-hydroxy-5-((E)-4-hydroxystyryl)phenoxy)-6-(hydroxym-
ethyl)tetrahydro-2H-pyran-3,4,5-triol; resveratrol; megestrol
acetate--chemical name
(8R,9S,10R,13S,14S,17R)-17-acetyl-6,10,13-trimethyl-3-oxo-2,3,8,9,10,11,1-
2,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-17-yl
acetate; docetaxel--chemical name
(2aR,4S,4aS,6R,9S,11R,12S,12aR,12bS)-12b-acetoxy-9-(((2R,3S)-3-((tert-but-
oxycarbonyl)amino)-2-hydroxy-3-phenylpropanoyl)oxy)-4,6,11-trihydroxy-4a,8-
,13,13-tetramethyl-5-oxo-2a,3,4,4a,5,6,9,10,11,12,12a,12b-dodecahydro-1H-7-
,11-methanocyclodeca[3,4]benzo[1,2-b]oxet-12-yl benzoate;
paclitaxel; perospirone--chemical name
(3aR,7aS)-2-(4-(4-(benzo[d]isothiazol-3-yl)piperazin-1-yl)butyl)hexahydro-
-1H-isoindole-1,3(2H)-dione hydrochloride; alprazolam--chemical
name
8-chloro-1-methyl-6-phenyl-4H-benzo[f][1,2,4]triazolo[4,3-a][1,4]diazepin-
e; diazepam; oxazepam; temazepam; lorazepam; clonazepam; midazolam;
fenoldopam--chemical name
6-chloro-1-(4-hydroxyphenyl)-2,3,4,5-tetrahydro-1H-benzo[d]azepine-7,8-di-
ol methanesulfonate; ondansetron--chemical name
9-methyl-3-((2-methyl-1H-imidazol-1-yl)methyl)-2,3-dihydro-1H-carbazol-4(-
9H)-one; letrozole--chemical name
4,4'-((1H-1,2,4-triazol-1-yl)methylene)dibenzonitrile;
taxifolin--chemical name
(2R,3S)-2-(3,4-dihydroxyphenyl)-3,5,7-trihydroxychroman-4-one;
cytarabine; methyltestosterone--chemical name
(8R,9S,10R,13S,14S,17S)-17-hydroxy-10,13,17-trimethyl-6,7,8,9,10,11,12,13-
,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-3 (2H)-one;
artesunate--chemical name
4-oxo-4-(((3R,5aS,6R,8aS,9R,10R,12R,12aR)-3,6,9-trimethyldecahydro-3H-3,1-
2-epoxy[1,2]dioxepino[4,3-i]isochromen-10-yl)oxy)butanoic acid;
artemether; dihydroartemisinin; artelinic acid; artenimol;
artemotil; triclabendaxole--chemical name
5-chloro-6-(2,3-dichlorophenoxy)-2-(methylthio)-1H-benzo[d]imidazole;
albendazole; mebendazole; thiabendazole; fenbendazole;
flubendazole; ezetimibe--chemical name
(3R,4S)-1-(4-fluorophenyl)-3-((S)-3-(4-fluorophenyl)-3-hydroxypropyl)-4-(-
4-hydroxyphenyl)azetidin-2-one; oxiconazole--chemical name
1-(2,4-dichlorophenyl)-2-(1H-imidazol-1-yl)ethanone
O-(2,4-dichlorobenzyl) oxime; gebexate--chemical name ethyl
4-((6-((diaminomethylene)amino) hexanoyl)oxy)benzoate;
risperidone--chemical name
3-(2-(4-(6-fluorobenzo[d]isoxazol-3-yl)piperidin-1-yl)ethyl)-2-methyl-6,7-
,8,9-tetrahydro-4H-pyrido[1,2-a]pyrimidin-4-one; paliperidone;
bifonazole--chemical name
1-([1,1'-biphenyl]-4-yl(phenyl)methyl)-1H-imidazole; indinavir;
calcitriol; vinorelbine; tegaserod; irbesartan; pterostilbene;
lovastatin; felodipine; daunorubicin hydrochloride; albendazole;
mitoxantrone; clomid; econazole nitrate; 5-fluorouracil; including
derivatives and those substituted with one or more substituents,
which are the same or different, or salts thereof. In certain
embodiments, bone graft compositions comprise compounds disclosed
herein and a bone morphogenetic protein and/or another growth
factor. Typically, the bone morphogenetic protein is BMP-2, BMP-5,
or BMP-7. In certain embodiments, the graft composition comprises
calcium phosphates and/or bone granules, hydroxyapatite and/or
beta-tricalcium phosphate, alpha-tricalcium phosphate,
polysaccharides or combinations thereof. Crushed bone granules,
typically obtained from the subject, are optionally added to the
graft composition.
[0009] In some embodiments, the graft contains osteogenic material
can be obtained from autogenic or allogenic sources and includes,
autograft, autogenic bone marrow aspirate, autogenic lipoaspirate,
allogenic bone marrow aspirate, allogenic lipoaspirate, and blends
and mixtures thereof.
[0010] In some embodiments, the disclosure relates to bone graft
compositions comprising a compound disclosed herein, such as
1-triarylmethyl-1H-imidazole derivatives, or salts thereof, such as
clotrimazole; biphenyl-diol derivatives, or salts thereof such as
honokiol and magnolol; macrolide lactone derivatives, or salts
thereof such as tacrolimus, pimecrolimus, sirolimus, everolimus,
temsirolimus; steroid derivatives, or salts thereof such as
spironolactone;
3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenant-
hrene-17-carbothioic acid derivatives, or salts thereof such as
fluticasone, fluticasone propionate, and fluticasone furoate;
3-(4-morpholinophenyl)oxazolidin-2-one derivatives, or salts
thereof such as linezolid; 1H,3'H-2,5'-bibenzo[d]imidazole
derivatives or salts thereof such as telmisartan; rentinol
derivatives, or salts thereof such as tretinoin, alitretinoin,
isotretinoin, retinol, etretinate, acitretin;
4-(4-(dialkylamino)phenyl) butanoic acid derivatives, or salts
thereof such as chlorambucil, podophyllotoxin derivatives such as
teniposide; aziridine derivatives such as mitomycin C; nucleoside
derivatives such as cytarabine, decitabine; vinca alkaloid
derivatives such as vinorelbine, vinblastine, vincristine, and
vindesine; anthracycline doxorubicin derivatives such as
doxorubicin, valrubicin, daunorubicin, epirubicin, idarubicin;
anthraquinone derivatives such as mitoxantrone and pixantrone;
plicamycin or derivatives; pazopanib or derivatives; camptothecin
or derivatives such as topotecan, irinotecan; sunitinib or
derivatives or salts thereof and a graft matrix. Typically, the
matrix comprises a collagen sponge and/or a compression resistant
type I collagen and calcium phosphates. In other embodiments, the
matrix is a hydrogel.
[0011] In certain embodiments, the disclosure contemplates a graft
composition or matrix comprising compounds disclosed herein such as
tacrolimus, pimecrolimus, sirolimus, everolimus, temsirolimus,
teniposide, mitomycin C, cytarabine, decitabine, vinblastine,
vincristine, vindesine, vinorelbine, valrubicin, doxorubicin,
daunorubicin, epirubicin, idarubicin, mitoxantrone, pixantrone,
plicamycin, pazopanib, topotecan, camptothecin, irinotecan,
sunitinib, wherein the graft matrix is a collagen or demineralized
bone matrix or ceramic of other scaffold disclosed herein without
exogenous cells.
[0012] In some embodiments, the disclosure relates to kits
comprising a graft composition, a compound disclosed herein, and a
graft matrix. In certain embodiments, the kits further comprise a
bone morphogenetic protein and/or another growth factor. In certain
embodiments, the kits further comprise a transfer device, such as a
syringe or pipette.
[0013] In some embodiments, the disclosure relates to methods of
generating BMP-mediated osteoblasts comprising administering an
effective amount of compound(s) disclosed herein to cells capable
of osteoblastic differentiation, such as mesenchymal stem cells and
pre-osteoblastic cells.
[0014] In some embodiments, the disclosure relates to methods of
forming bone or cartilage, comprising implanting a graft
composition comprising a compound disclosed herein, such as
1-triarylmethyl-1H-imidazole derivatives, or salts thereof, such as
clotrimazole; biphenyl-diol derivatives, or salts thereof such as
honokiol and magnolol; macrolide lactone derivatives, or salts
thereof such as tacrolimus, pimecrolimus, sirolimus, everolimus,
temsirolimus; steroid derivatives, or salts thereof such as
spironolactone;
3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenant-
hrene-17-carbothioic acid derivatives, or salts thereof such as
fluticasone, fluticasone propionate, and fluticasone furoate;
3-(4-morpholinophenyl)oxazolidin-2-one derivatives, or salts
thereof such as linezolid; 1H,3'H-2,5'-bibenzo[d]imidazole
derivatives or salts thereof such as telmisartan; rentinol
derivatives, or salts thereof such as tretinoin, alitretinoin,
isotretinoin, retinol, etretinate, acitretin;
4-(4-(dialkylamino)phenyl)butanoic acid derivatives, or salts
thereof such as chlorambucil, podophyllotoxin derivatives such as
teniposide; aziridine derivatives such as mitomycin C; nucleoside
derivatives such as cytarabine, decitabine; vinca alkaloid
derivatives such as vinorelbine, vinblastine, vincristine, and
vindesine; anthracycline doxorubicin derivatives such as
doxorubicin, valrubicin, daunorubicin, epirubicin, idarubicin;
anthraquinone derivatives such as mitoxantrone and pixantrone;
plicamycin or derivatives; pazopanib or derivatives; camptothecin
or derivatives such as topotecan, irinotecan; sunitinib or
derivatives or salts thereof in a subject under conditions such
that bone or cartilage forms in the graft. Typically, the subject
has a void in the bony structure wherein the graft composition is
implanted in the void. In certain embodiments, the void is in a
bone selected from an extremity, maxilla, mandible, pelvis, spine
and/or cranium. In certain embodiments, the void is a result of
surgical removal of bone. In certain embodiments, the void is
between bone and an implanted medical device. In another
embodiment, the method further comprises the step of securing
movement of bone structure with a fixation system, and removing the
system after bone forms in the implanted graft.
[0015] In certain embodiments, the disclosure contemplates local
bone formation for fracture repair, segmental bone defects, spine
fusion, bone grafting, and regional bone enhancement for osteopenic
bones before they fracture (e.g. hip, vertebral body, etc) by
deliver locally to induce local bone formation.
[0016] In certain embodiments, the disclosure relates to methods of
growing bone in subject by locally administering, such as by
injection, a composition comprising a compound disclosed herein,
optionally in combination with a growth factor, about the area of
desired bone growth. In certain embodiments, the disclosure relates
to methods of growing bone comprising administering a
pharmaceutical composition comprising clotrimazole, honokiol,
magnolol, tacrolimus, pimecrolimus, sirolimus, everolimus,
temsirolimus, spironolactone, fluticasone, fluticasone propionate,
fluticasone furoate, linezolid, telmisartan, chlorambucil, retinol,
isotretinoin, acitretin, etretinate, retinoic acid (tretinoin),
teniposide, mitomycin C, cytarabine, decitabine, vinblastine,
vincristine, vindesine, vinorelbine, valrubicin, doxorubicin,
daunorubicin, epirubicin, idarubicin, mitoxantrone, pixantrone,
plicamycin, pazopanib, topotecan, camptothecin, irinotecan,
sunitinib, derivatives, or pharmaceutically acceptable salts
thereof to a subject in an area of desired growth, wherein the
administration is localized directly about the area of desired
growth. In certain embodiments, the administration is not oral
administration. In certain embodiments, the administration is
through a catheter or hypodermic needle with a tip that is not in a
vein. In certain embodiments, the administration is by injection
into the subcutaneous tissue or in or about an area typically
occupied by bone or between vertebra, e.g., in the area usually
occupied by in the intervertebral disc, to form a spinal
fusion.
[0017] In certain embodiments, the method contemplates implanting a
graft composition in a desired area of the subject and locally
administering a composition comprising a compound disclosed herein,
optionally in combination with a growth factor, in the graft or
about the area of the graft implant such as by injection.
[0018] In certain embodiments, the disclosure relates to uses of
compounds disclosed herein for cartilage regeneration e.g., between
intervertebral disc and articular, jaw, elbow, knee, ankle, wrist,
and hip joints. Methods contemplate oral administration,
intravenous administration, or direct injection at the desired
site(s) of the subject.
[0019] In some embodiments, the disclosure relates to methods of
performing spinal fusion comprising implanting a bone graft
composition comprising a compound disclosed herein, such as
1-triarylmethyl-1H-imidazole derivatives, or salts thereof, such as
clotrimazole; biphenyl-diol derivatives, or salts thereof such as
honokiol and magnolol; macrolide lactone derivatives, or salts
thereof such as tacrolimus, pimecrolimus, sirolimus, everolimus,
temsirolimus; steroid derivatives, or salts thereof such as
spironolactone;
3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenant-
hrene-17-carbothioic acid derivatives, or salts thereof such as
fluticasone, fluticasone propionate, and fluticasone furoate;
3-(4-morpholinophenyl)oxazolidin-2-one derivatives, or salts
thereof such as linezolid; 1H,3'H-2,5'-bibenzo[d]imidazole
derivatives or salts thereof such as telmisartan; rentinol
derivatives, or salts thereof such as tretinoin, alitretinoin,
isotretinoin, retinol, etretinate, acitretin;
4-(4-(dialkylamino)phenyl)butanoic acid derivatives, or salts
thereof such as chlorambucil, podophyllotoxin derivatives such as
teniposide; aziridine derivatives such as mitomycin C; nucleoside
derivatives such as cytarabine, decitabine; vinca alkaloid
derivatives such as vinorelbine, vinblastine, vincristine, and
vindesine; anthracycline doxorubicin derivatives such as
doxorubicin, valrubicin, daunorubicin, epirubicin, idarubicin;
anthraquinone derivatives such as mitoxantrone and pixantrone;
plicamycin or derivatives; pazopanib or derivatives; camptothecin
or derivatives such as topotecan, irinotecan; sunitinib or
derivatives or salts thereof configured to grow bone between two
vertebrae of a subject. In certain embodiments, the composition
further comprises a bone morphogenetic protein and/or another
growth factor. In a typical embodiment, the subject is diagnosed
with degenerative disc disease or has symptoms of back pain.
[0020] In some embodiments, the disclosure relates to methods of
inserting a prosthetic device or anchor, comprising exposing a bone
and implanting a graft composition comprising a compound disclosed
herein in contact with the bone. In certain embodiments, one
implants the prosthetic device or anchor in the graft composition.
In certain embodiments, the composition further comprises a bone
morphogenetic protein and/or another growth factor.
[0021] In some embodiments, the disclosure relates to
pharmaceutical compositions comprising compounds disclosed herein
or pharmaceutically acceptable salts thereof. In certain
embodiments, the composition further comprises a bone morphogenetic
protein and/or another growth factor. In certain embodiments, the
pharmaceutical composition is formulated to release over a 12 hour,
1 day, 3 day, 5 day, 7 day, two week, or one month period.
[0022] In certain embodiments, the disclosure relates to methods of
preventing or treating a bone fracture, comprising administering a
pharmaceutical composition comprising a compound disclosed herein,
such as 1-triarylmethyl-1H-imidazole derivatives, or salts thereof,
such as clotrimazole; biphenyl-diol derivatives, or salts thereof
such as honokiol and magnolol; macrolide lactone derivatives, or
salts thereof such as tacrolimus, pimecrolimus, sirolimus,
everolimus, temsirolimus; steroid derivatives, or salts thereof
such as spironolactone;
3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenant-
hrene-17-carbothioic acid derivatives, or salts thereof such as
fluticasone, fluticasone propionate, and fluticasone furoate;
3-(4-morpholinophenyl) oxazolidin-2-one derivatives, or salts
thereof such as linezolid; 1H,3'H-2,5'-bibenzo[d]imidazole
derivatives or salts thereof such as telmisartan; rentinol
derivatives, or salts thereof such as tretinoin, alitretinoin,
isotretinoin, retinol, etretinate, acitretin;
4-(4-(diethylamino)phenyl)butanoic acid derivatives, or salts
thereof such as chlorambucil; podophyllotoxin derivatives such as
teniposide; aziridine derivatives such as mitomycin C; nucleoside
derivatives such as cytarabine, decitabine; vinca alkaloid
derivatives such as vinorelbine, vinblastine, vincristine, and
vindesine; anthracycline doxorubicin derivatives such as
doxorubicin, valrubicin, daunorubicin, epirubicin, idarubicin;
anthraquinone derivatives such as mitoxantrone and pixantrone;
plicamycin or derivatives; pazopanib or derivatives; camptothecin
or derivatives such as topotecan, irinotecan; sunitinib or
derivatives or salts thereof or a pharmaceutically acceptable salt
thereof, to a subject at risk for, exhibiting symptoms of, or
diagnosed with a bone fracture. In certain embodiments, the
composition further comprises a bone morphogenetic protein and/or
another growth factor.
[0023] In certain embodiments, the administration is localized. In
certain embodiments, administration is achieved through oral
delivery, intravenous delivery, parenteral delivery, intradermal
delivery, percutaneous delivery, or subcutaneous delivery. In some
embodiments, the method further comprises the step of exposing the
bone fracture to pulsed electromagnetic fields. In further
embodiments, the subject is diagnosed with a long bone shaft
fracture such as a tibia or femur fracture corrected with
intramedullary nail fixation.
[0024] In some embodiments, the disclosure relates to methods of
preventing or treating a bone degenerative disease, comprising
administering a pharmaceutical composition comprising a compound
disclosed herein, such as 1-triarylmethyl-1H-imidazole derivatives,
or salts thereof, such as clotrimazole; biphenyl-diol derivatives,
or salts thereof such as honokiol and magnolol; macrolide lactone
derivatives, or salts thereof such as tacrolimus, pimecrolimus,
sirolimus, everolimus, temsirolimus; steroid derivatives, or salts
thereof such as spironolactone;
3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenant-
hrene-17-carbothioic acid derivatives, or salts thereof such as
fluticasone, fluticasone propionate, and fluticasone furoate;
3-(4-morpholinophenyl) oxazolidin-2-one derivatives, or salts
thereof such as linezolid; 1H,3'H-2,5'-bibenzo[d]imidazole
derivatives or salts thereof such as telmisartan; rentinol
derivatives, or salts thereof such as tretinoin, alitretinoin,
isotretinoin, retinol, etretinate, acitretin;
4-(4-(diethylamino)phenyl) butanoic acid derivatives, or salts
thereof such as chlorambucil; podophyllotoxin derivatives such as
teniposide; aziridine derivatives such as mitomycin C; nucleoside
derivatives such as cytarabine, decitabine; vinca alkaloid
derivatives such as vinorelbine, vinblastine, vincristine, and
vindesine; anthracycline doxorubicin derivatives such as
doxorubicin, valrubicin, daunorubicin, epirubicin, idarubicin;
anthraquinone derivatives such as mitoxantrone and pixantrone;
plicamycin or derivatives; pazopanib or derivatives; camptothecin
or derivatives such as topotecan, irinotecan; sunitinib or
derivatives or salts thereof or a pharmaceutically acceptable salts
thereof, to a subject at risk for, exhibiting symptoms of, or
diagnosed with a bone degenerative disease. In certain embodiments,
the composition further comprises a bone morphogenetic protein
and/or another growth factor. In certain embodiments, the
administration is systemic, or administration is achieved through
oral delivery, intravenous delivery, parenteral delivery,
intradermal delivery, percutaneous delivery, or subcutaneous
delivery. In some embodiments, the disease is osteoporosis,
osteitis deformans, bone metastasis, multiple myeloma, primary
hyperparathyroidism, or osteogenesis imperfecta.
[0025] In some embodiments, the disclosure relates to methods for
decreasing the time required to form new bone in the presence of a
bone morphogenetic protein, comprising co-administering at least
one compound disclosed herein, such as 1-triarylmethyl-1H-imidazole
derivatives, or salts thereof, such as clotrimazole; biphenyl-diol
derivatives, or salts thereof such as honokiol and magnolol;
macrolide lactone derivatives, or salts thereof such as tacrolimus,
pimecrolimus, sirolimus, everolimus, temsirolimus; steroid
derivatives, or salts thereof such as spironolactone;
3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenant-
hrene-17-carbothioic acid derivatives, or salts thereof such as
fluticasone, fluticasone propionate, and fluticasone furoate;
3-(4-morpholinophenyl)oxazolidin-2-one derivatives, or salts
thereof such as linezolid; 1H,3'H-2,5'-bibenzo[d]imidazole
derivatives or salts thereof such as telmisartan; rentinol
derivatives, or salts thereof such as tretinoin, alitretinoin,
isotretinoin, retinol, etretinate, acitretin;
4-(4-(dialkylamino)phenyl)butanoic acid derivatives, or salts
thereof such as chlorambucil; podophyllotoxin derivatives such as
teniposide; aziridine derivatives such as mitomycin C; nucleoside
derivatives such as cytarabine, decitabine; vinca alkaloid
derivatives such as vinorelbine, vinblastine, vincristine, and
vindesine; anthracycline doxorubicin derivatives such as
doxorubicin, valrubicin, daunorubicin, epirubicin, idarubicin;
anthraquinone derivatives such as mitoxantrone and pixantrone;
plicamycin or derivatives; pazopanib or derivatives; camptothecin
or derivatives such as topotecan, irinotecan; sunitinib or
derivatives or salts thereof and another active ingredient.
[0026] In some embodiments, the disclosure relates to a process for
engineering bone tissue comprising combining a compound disclosed
herein, such as 1-triarylmethyl-1H-imidazole derivatives, or salts
thereof, such as clotrimazole; biphenyl-diol derivatives, or salts
thereof such as honokiol and magnolol; macrolide lactone
derivatives, or salts thereof such as tacrolimus, pimecrolimus,
sirolimus, everolimus, temsirolimus; steroid derivatives, or salts
thereof such as spironolactone;
3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenant-
hrene-17-carbothioic acid derivatives, or salts thereof such as
fluticasone, fluticasone propionate, and fluticasone furoate;
3-(4-morpholinophenyl)oxazolidin-2-one derivatives, or salts
thereof such as linezolid; 1H,3'H-2,5'-bibenzo[d]imidazole
derivatives or salts thereof such as telmisartan; rentinol
derivatives, or salts thereof such as tretinoin, alitretinoin,
isotretinoin, retinol, etretinate, acitretin;
4-(4-(dialkylamino)phenyl)butanoic acid derivatives, or salts
thereof such as chlorambucil; podophyllotoxin derivatives such as
teniposide; aziridine derivatives such as mitomycin C; nucleoside
derivatives such as cytarabine, decitabine; vinca alkaloid
derivatives such as vinorelbine, vinblastine, vincristine, and
vindesine; anthracycline doxorubicin derivatives such as
doxorubicin, valrubicin, daunorubicin, epirubicin, idarubicin;
anthraquinone derivatives such as mitoxantrone and pixantrone;
plicamycin or derivatives; pazopanib or derivatives; camptothecin
or derivatives such as topotecan, irinotecan; sunitinib or
derivatives or salts thereof and optionally a bone morphogenetic
protein, with a cell selected from the group consisting of
osteogenic cells, pluripotent stem cells, mesenchymal cells, and
embryonic stem cells.
[0027] In certain embodiments, the disclosure relates to using
1-triarylmethyl-1H-imidazole derivatives, or salts thereof, such as
clotrimazole; biphenyl-diol derivatives, or salts thereof such as
honokiol and magnolol; macrolide lactone derivatives, or salts
thereof such as tacrolimus, pimecrolimus, sirolimus, everolimus,
temsirolimus; steroid derivatives, or salts thereof such as
spironolactone;
3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenant-
hrene-17-carbothioic acid derivatives, or salts thereof such as
fluticasone, fluticasone propionate, and fluticasone furoate;
3-(4-morpholinophenyl)oxazolidin-2-one derivatives, or salts
thereof such as linezolid; 1H,3'H-2,5'-bibenzo[d]imidazole
derivatives or salts thereof such as telmisartan; rentinol
derivatives, or salts thereof such as tretinoin, alitretinoin,
isotretinoin, retinol, etretinate, acitretin;
4-(4-(dialkylamino)phenyl)butanoic acid derivatives, or salts
thereof such as chlorambucil; podophyllotoxin derivatives such as
teniposide; aziridine derivatives such as mitomycin C; nucleoside
derivatives such as cytarabine, decitabine; vinca alkaloid
derivatives such as vinorelbine, vinblastine, vincristine, and
vindesine; anthracycline doxorubicin derivatives such as
doxorubicin, valrubicin, daunorubicin, epirubicin, idarubicin;
anthraquinone derivatives such as mitoxantrone and pixantrone;
plicamycin or derivatives; pazopanib or derivatives; camptothecin
or derivatives such as topotecan, irinotecan; sunitinib or
derivatives or salts thereof in the production of a medicament for
the treatment or prevention of a bone disease or other applications
disclosed herein.
[0028] In certain embodiments, the disclosure relates to a bone
graft composition comprising a bone growth-inducing amount of
1-triarylmethyl-1H-imidazole derivatives, or salts thereof, such as
clotrimazole; biphenyl-diol derivatives, or salts thereof such as
honokiol and magnolol; macrolide lactone derivatives, or salts
thereof such as tacrolimus, pimecrolimus, sirolimus, everolimus,
temsirolimus; steroid derivatives, or salts thereof such as
spironolactone;
3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenant-
hrene-17-carbothioic acid derivatives, or salts thereof such as
fluticasone, fluticasone propionate, and fluticasone furoate;
3-(4-morpholinophenyl)oxazolidin-2-one derivatives, or salts
thereof such as linezolid; 1H,3'H-2,5'-bibenzo[d]imidazole
derivatives or salts thereof such as telmisartan; rentinol
derivatives, or salts thereof such as tretinoin, alitretinoin,
isotretinoin, retinol, etretinate, acitretin;
4-(4-(diethylamino)phenyl) butanoic acid derivatives, or salts
thereof such as chlorambucil; podophyllotoxin derivatives such as
teniposide; aziridine derivatives such as mitomycin C; nucleoside
derivatives such as cytarabine, decitabine; vinca alkaloid
derivatives such as vinorelbine, vinblastine, vincristine, and
vindesine; anthracycline doxorubicin derivatives such as
doxorubicin, valrubicin, daunorubicin, epirubicin, idarubicin;
anthraquinone derivatives such as mitoxantrone and pixantrone;
plicamycin or derivatives; pazopanib or derivatives; camptothecin
or derivatives such as topotecan, irinotecan; sunitinib or
derivatives or salts thereof and a pharmaceutically acceptable
carrier.
[0029] In certain embodiments, the derivative is
1-triarylmethyl-1H-imidazole or salt thereof with one or more
substituents.
[0030] In certain embodiments, the derivative is
1,1'-biphenyl-2,4'-diol or salt thereof with one or more
substituents.
[0031] In certain embodiments, the derivative is
5,6,8,11,12,13,14,15,16,17,18,19,24,25,26,26a-hexadecahydro-3-[(1E)-2-[cy-
clohexyl]ethenyl]-15,19-epoxy-3H-pyrido[2,1-c][1,4]oxaazacyclotricosine-1,-
7,20,21(4H,23H)-tetrone or salt thereof with one or more
substituents.
[0032] In certain embodiments, the derivative is
3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenant-
hrene-17-carbothioic acid or salt thereof with one or more
substituents.
[0033] In certain embodiments, the derivative is
1,6,7,8,9,10,11,12,13,14,15,16-dodecahydro-3'H-spiro[cyclopenta[a]phenant-
hrene-17,2'-furan]-3,5'(2H,4'H)-dione or salt thereof with one or
more substituents.
[0034] In certain embodiments, the derivative is
3-(4-morpholinophenyl)oxazolidin-2-one or salt thereof with one or
more substituents.
[0035] In certain embodiments, the derivative is
1-(octa-1,3,5,7-tetraen-1-yl)cyclohex-1-ene or
octa-1,3,5,7-tetraen-1-ylbenzene or salt thereof with one or more
substituents.
[0036] In certain embodiments, the derivative is
3'-([1,1'-biphenyl]-4-ylmethyl)-1H,3'H-2,5'-bibenzo[d]imidazole or
salt thereof with one or more substituents.
[0037] In certain embodiments, the derivative is
4-(4-(dialkylamino)phenyl)butanoic acid or salt thereof with one or
more substituents.
[0038] In certain embodiments, the 1-triarylmethyl-1H-imidazole
derivative is clotrimazole or salt thereof optionally substituted
with one or more substituents.
[0039] In certain embodiments, the biphenyl-diol derivative is
honokiol or magnolol or salt thereof optionally substituted with
one or more substituents.
[0040] In certain embodiments, the macrolide lactone derivative is
tacrolimus, pimecrolimus, sirolimus, everolimus, temsirolimus or
salt thereof optionally substituted with one or more
substituents.
[0041] In certain embodiments, the
3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenant-
hrene-17-carbothioic acid derivative is fluticasone, fluticasone
propionate, or fluticasone furoate or salt thereof optionally
substituted with one or more substituents.
[0042] In certain embodiments, the
3-(4-morpholinophenyl)oxazolidin-2-one derivative is linezolid or
salt thereof optionally substituted with one or more
substituents.
[0043] In certain embodiments, the 1H,3'H-2,5'-bibenzo[d]imidazole
derivative is telmisartan or salt thereof optionally substituted
with one or more substituents.
[0044] In certain embodiments, the steroid derivative is
spironolactone or salt thereof optionally substituted with one or
more substituents.
[0045] In certain embodiments, the retinol derivative is tretinoin,
alitretinoin, isotretinoin, retinol, etretinate, acitretin or salt
thereof optionally substituted with one or more substituents.
[0046] In certain embodiments, the
4-(4-(dialkylamino)phenyl)butanoic acid derivative is chlorambucil
or salt thereof with one or more substituents.
[0047] In certain embodiments, the podophyllotoxin is
5-phenyl-9-((2-(thiophen-2-yl)hexahydropyrano[3,2-d][1,3]dioxin-6-yl)oxy)-
-5,5a,8a,9-tetrahydrofuro[3',4':6,7]naphtho[2,3-d][1,3]dioxol-6(8H)-one
or derivative such as teniposide optionally substituted with one or
more substituents.
[0048] In certain embodidments, the aziridine is
1,1a,2,8,8a,8b-hexahydroazirino[2',3':3,4]pyrrolo[1,2-a]indole-4,7-dione
or derivative such as mitomycin C optionally substituted with one
or more substituents.
[0049] In certain embodiments, the nucleoside is
1-(5-methyltetrahydrofuran-2-yl)pyrimidin-2(1H)-one or derivative
such as cytarabine, decitabine optionally substituted with one or
more substituents.
[0050] In certain embodiments, the vinca alkaloid is
9-(2,4,5,6,7,8,9,10-octahydro-1H-3,7-methano[1]azacycloundecino[5,4-b]ind-
ol-9-yl)-3a,3a1,4,5,5a,6,11,12-octahydro-1H-indolizino[8,1-cd]carbazole
or derivative such as vinorelbine, vinblastine, vincristine, and
vindesine optionally substituted with one or more substituents.
[0051] In certain embodiments, the anthracycline is
8-acetyl-10-((4-amino-5-hydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)-6,8-
,11-trihydroxy-1-methoxy-7,8,9,10-tetrahydrotetracene-5,12-dione or
derivative such as doxorubicin, valrubicin, daunorubicin,
epirubicin, idarubicin optionally substituted with one or more
substituents.
[0052] In certain embodiments, the anthraquinone is
anthracene-9,10-dione or derivative such as mitoxantrone and
pixantrone optionally substituted with one or more
substituents.
[0053] In certain embodiments, plicamycin or derivatives are
optionally substituted with one or more substituents.
[0054] In certain embodiment, pazopanib or derivatives are
optionally substituted with one or more substituents.
[0055] In certain embodiments, the camptothecin is
1H-pyrano[3',4':6,7]indolizino[1,2-b]quinoline-3,14(4H,12H)-dione
or derivatives such as topotecan and irinotecan optionally
substituted with one or more substituents.
[0056] In certain embodiments, sunitinib or derivatives are
optionally substituted with one or more substituents.
[0057] In certain embodiments, the bone morphogenetic protein is
selected from the group consisting BMP-2, BMP-5, BMP-6, BMP-7,
BMP-9, and combinations thereof.
[0058] In certain embodiments, the compounds described herein may
be used locally such as injection percutaneously at any bone
formation site (fracture, spine fusion delayed a day or several
days after surgery). In certain embodiments, the compounds may also
be bound to a matrix or scaffold and delivered with growth factors,
cells (MSCs or others), or on a dry carrier matrix to direct local
bone formation in the shape of the carrier/scaffold. Within certain
embodiments, it is also contemplated that one or more of these
compounds disclosed herein may be used alone or in combination with
multiple compounds, with or without exogenous growth factors,
and/or in combination with other promoting agents of the BMP
pathway such as a noggin inhibitor, a Smurf inhibitor and/or a JAB
1 inhibitor.
[0059] In certain embodiments, the disclosure contemplates delivery
of compositions comprising compounds disclosed herein via a liquid
or flowable gel optionally including collagen, hydroxyapatite,
demineralized bone, or polymer matrix or others, e.g., as disclosed
herein. In certain embodiments, the compositions are injected into
the central cavity or interstices of a structural element such as a
bone cage made from allograft, polymer thermoplastic, metal such as
titanium or aluminum, polyether ether ketone (PEEK) such as those
generated from 4,4'-difluorobenzophenone, or other polymer.
[0060] In certain embodiments, the disclosure contemplates bone
grafts containing more than 10, 15, 20, 25, 30, 35 mM of a compound
disclosed herein such as a macrolide lactone derivative such as
tacrolimus, pimecrolimus, sirolimus, everolimus, temsirolimus per
about 100 mm.sup.3, or about 150 mm.sup.3, or 50 to 200 mm.sup.3,
or about 100 to 200 mm.sup.3 of bone graft volume. In some
embodiments, the disclosure contemplates more than 10 mM or 20 mM
of sirolimus. In some embodiments, the disclosure contemplates more
than 10 mM or 15 mM of everolimus. In some embodiments, the
disclosure contemplates more than 10 mM or 15 mM of tacrolimus. In
certain embodiments, the bone graft is contemplated for all uses
disclose herein. In certain embodiments, the subject is human.
[0061] In certain embodiments, the disclosure contemplates bone
graft containing more than 0.7, 0.8, 0.9, 1.0, 1.2, 1.5, 1.8, 2.0,
or 3.0 mg of a macrolide lactone derivative such as tacrolimus,
pimecrolimus, sirolimus, everolimus, temsirolimus, per about 100
mm.sup.3, or about 150 mm.sup.3, or 50 to 200 mm.sup.3, or about
100 to 200 mm.sup.3 of bone graft volume. In some embodiments, the
disclosure contemplates more than 1.5 mg of sirolimus. In some
embodiments, the disclosure contemplates more that 0.9 mg of
everolimus. In some embodiments, the disclosure contemplates more
that 0.7 mg of tacrolimus. In certain embodiments, the bone graft
is contemplated for all uses disclose herein. In certain
embodiments, the subject is human.
[0062] In certain embodiments, the disclosure contemplates kit
containing a bone graft and a composition comprising more than 0.7,
0.8, 0.9, 1.0, 1.2, 1.5, 1.8, 2.0, or 3.0 mg of a macrolide lactone
derivative such as tacrolimus, pimecrolimus, sirolimus, everolimus,
temsirolimus per about 100, 125, or 150 mm.sup.3 of bone graft
volume. In some embodiments, the disclosure contemplates more than
1.5 mg of sirolimus. In some embodiments, the disclosure
contemplates more that 0.9 mg of everolimus. In some embodiments,
the disclosure contemplates more that 0.7 mg of tacrolimus.
BRIEF DESCRIPTION OF THE FIGURES
[0063] FIG. 1 illustrates the BMP signaling pathway. Although it is
not intended that embodiments of the disclosure be limited by any
particular mechanism, it is believed that the embodiments may act
through the BMP pathway. In the BMP pathway, BMP binds to the BMP
receptor type II that in turn activates BMPR-1. Activated BMPR-1
phosphorylates receptor regulated Smads (Smad1/5/8) which form
complexes with Smad4 facilitating nuclear entry of Smad complexes.
Activated Smad complexes regulate gene expression of BMP-responsive
genes. Extracellularly, Noggin antagonizes signaling by binding to
BMP and thus inhibiting binding of BMP to its receptor complex.
[0064] FIG. 2 illustrates the structural features of BMP-Receptor
interactions and important amino acids that determine binding
specificity and affinity. Illustrative structures of BMP-2,
BMPR-IA, and BMPR-II are depicted. Important aminoacids are labeled
in `yellow` color.
[0065] FIG. 3 shows data for the activity of test compounds in the
ALP assay. Capsaicin is SNGP-003-01 H04; diphenylcyclopropenone is
SNGP-003-01 E05; clotrimazole is SNGP-003-01 C06; beta-estradiol is
SNGP-003-01 B09; ketoconazole or
1-(4-(4-(((2R,4S)-2-((1H-imidazol-1-yl)methyl)-2-(2,4-dichlorophenyl)-1,3-
-dioxolan-4-yl)methoxy)phenyl)piperazin-1-yl)ethanone is
SNGP-003-01 A10; and ethynylestradiol is SNGP-003-01 E10.
[0066] FIG. 4 shows data on the activity of test compounds in a
BMP-Noggin competitive ALP assay. Progesterone is SNGP-003-01 H03;
capsaicin is SNGP-003-01 H04; salbutamol sulfate is SNGP-003-01
A05; Zardaverine is SNGP-003-01 H05; clotrimazole is SNGP-003-01
C06; Riluzole is SNGP-003-01 F06; (CGS 15943)
9-chloro-2-(furan-2-yl)-[1,2,4]triazolo[1,5-c]quinazolin-5-amine is
SNGP-003-01 B07; Prazosin is SNGP-003-01 G07; and urapidil
hydrochloride is SNGP-003-01 H07. (CGS 15943)
9-chloro-2-(furan-2-yl)-[1,2,4]triazolo[1,5-c]quinazolin-5-amine
and Capsaicin caused cell lifting from culture plates as determined
based on low cell number and low yield of protein after cell
harvesting and cell lysis.
[0067] FIG. 5 shows data suggesting dose dependent increase of
BMP-induced ALP activity by nanomolar concentrations of
clotrimazole.
[0068] FIG. 6 shows data for the activity of test compounds in the
ALP assay. Docetaxel is SNGP-003-02 A06; honokiol is SNGP-003-02
B06; carmofur is SNGP-003-02 D06; Methyltestosterone is SNGP-003-02
A07; itraconazole is SNGP-003-02 F07; triclabendazole is
SNGP-003-02 C08; flubendazole is SNGP-003-02 A09; tacrolimus is
SNGP-003-02 B09; and trimebutine maleate is SNGP-003-02 F09.
Docetaxel, carmofur, itraconazole, and flubendazole caused cell
lifting from culture plates.
[0069] FIG. 7 shows data of activity for test compounds in a
BMP-Noggin competitive ALP assay. Ketoconazole is SNGP-003-01 A10;
ethynylestradiol is SNGP-003-01 E10; cytarabine is SNGP-003-01 F10;
taxifolin is SNGP-003-02 E03; letrozole is SNGP-003-02 C04;
ondansetron is SNGP-003-02 F04; alprazolam is SNGP-003-02 A05;
perospirone is SNGP-003-02 H05; docetaxel is SNGP-003-02 A06; and
honokiol is SNGP-003-02 B06. Ketoconazole and cytarabine caused
cell lifting from culture plates.
[0070] FIG. 8 shows data on the activity of test compounds in a
BMP-Noggin competitive ALP Assay. Methyltestosterone is SNGP-003-02
A07; artesunate is SNGP-003-02 E07; triclabendazole is SNGP-003-02
C08; ezetimibe is SNGP-003-02 F08; tacrolimus is SNGP-003-02 B09;
trimebutine maleate is SNGP-003-02 F09; megestrol acetate is
SNGP-003-02 E10; and oxiconazole nitrate is SNGP-003-02 B11.
Artesunate and oxiconazole nitrate caused cell lifting from culture
plates.
[0071] FIG. 9 shows data on the activity of test compounds in an
ALP assay. Oxiconazole nitrate is SNGP-003-02 B11; pioglitazone
hydrochloride is SNGP-003-03 E04; zolpidem tartarate is SNGP-003-03
D05; fenoldopam mesylate is SNGP-003-03 H05; 2',3'-dideoxyinosine
is SNGP-003-03 E06; calcipotriol is SNGP-003-03 G06; icariin is
SNGP-003-03 E07; rosiglitazole hydrochloride is SNGP-003-03 H07;
oligomycin is SNGP-003-03 B08; rolipram is SNGP-003-03 G08;
fluticasone propionate is SNGP-003-03 G10; indinavir sulphate is
SNGP-003-03 H10; midazolam hydrochloride is SNGP-003-03 A11;
bifonazole is SNGP-003-04 CO2; calcitriol is SNGP-003-04 CO3;
vinorelbinee bitatrate is SNGP-003-04 B04; linezolid is SNGP-003-04
C04; irbesartan is SNGP-003-04 F04; and pterostilbene is
SNGP-003-04 A05. Calcipotriol, calcitriol, and vinorelbinee
bitatrate caused cell lifting from culture plates.
[0072] FIG. 10 shows data on the activities in ALP assays for
select compounds at 250 nM concentration. SVAK12X is
cycloguanil--chemical name
1-(4-chlorophenyl)-2,2-dimethyl-1,3,5-triazine-4,6-diamine.
[0073] FIG. 11 shows data on the activity of test compounds in a
BMP-Noggin competitive ALP assay. Telmisartan is
SNGP-003-04H08;
[0074] FIG. 12 shows data on the activity of test compounds in an
ALP assay. Retinoic acid is SNGP-201-01-006; spironolactone is
SNGP-201-01D06; 5-fluorouracil is SNGP-201-01-H02;
3,5,3'-triiodothryonine is SNGP-201-01-A08; econazole nitrate is
SNGP-201-01-E09; oxytetracycline hydrochloride is SNGP-201-01-H11.
Econazole nitrate and 5-fluorouracil caused cell lifting from
culture plates.
[0075] FIG. 13 shows data on the activity of test compounds in an
ALP assay. Chlorambucil is SNGP-201-02-E05; isotretinoin is
SNGP-201-03-E05; albendazole is SNGP-201-03-A03; mitoxantrone is
SNGP-201-03-B04; clomid is SNGP-201-03-A05. Albendazole,
mitoxantrone, and clomid caused cell lifting from culture
plates.
[0076] FIG. 14 shows data of activity for test compounds in a
BMP-Noggin competitive ALP assay. Isotretinoin is
SNGP-201-03-E05.
[0077] FIG. 15 shows data on the activity of test compounds in an
ALP assay. Isotretinoin is SNGP-201-03-E05 and acitretin is
SNGP-201-04-E06. Daunorubicin hydrochloride is SNGP-201-04-E03.
Daunorubicin hydrochloride caused cell lifting from culture
plates.
[0078] FIG. 16 shows data of activity for test compounds in a
BMP-Noggin competitive ALP assay. Acitretin is SNGP-201-04-E06;
lovastatin is SNGP-201-04-CO5; and felodipine is SNGP-201-04-006.
Lovastatin and felodipine caused cell lifting from culture
plates.
[0079] FIG. 17 shows data of activity for macrolide analogs in an
ALP activity assay. Vertical axis (Y-axis) is relative ALP
activity, and the horizontal axis (X-axis) is treatment.
DETAILED DISCUSSION
Terms
[0080] "Ossification" refers to the process of laying down new bone
by cells called osteoblasts. The term includes the growth in
healing bone fractures treated by cast or by open reduction and
stabilization by metal plate and screws. Ossification can also
result in the formation of bone tissue in an extraskeletal
location.
[0081] The term "bone morphogenetic protein" or "BMP" refers to any
one of the family of growth factors or fragments thereof with the
ability to induce the formation of bone and/or cartilage. The BMP
receptors are typically serine-threonine kinases. It is not
intended that BMP refer to any particular protein sequence and may
or may not have certain glycosylation patterns attached thereto
provided that the molecule has sufficient structural homology to
any one of the known BMPs described below and retains some
functional ability to promote bone growth, cartilage growth, or
osteoblast differentiation. BMPs may be isolated from natural or
non-natural sources, such as, but not limited to, recombinant or
synthetic methods. References to BMPs generally or a specific BMP,
e.g, BMP-2, includes recombinant or synthetically isolated versions
unless otherwise provide for herein. Combinations of BMPs are
contemplated. BMP-2 is known to induce bone and cartilage formation
and play a role in osteoblast differentiation. BMP-3 is known to
induce bone formation. BMP-4 is known to regulate the formation of
teeth, limbs and bone from mesoderm and play a role in fracture
repair. BMP-5 is known to function in cartilage development. BMP-6
is known to play a role in joint integrity and bone
formation/repair. BMP-7 and BMP-9 are known to play a role in
osteoblast differentiation. BMP-1 is a known metalloprotease that
acts on procollagen I, II, and III and is involved in cartilage
development.
[0082] As used herein, the term "derivative" refers to a
structurally similar compound that retains sufficient functional
attributes of the identified analogue. The derivative may be
structurally similar because it is lacking one or more atoms,
substituted, a salt, in different hydration/oxidation states, or
because one or more atoms within the molecule are switched, such
as, but not limited to, replacing an oxygen atom with a sulfur atom
or replacing an amino group with a hydroxy group. The derivative
may be a prodrug. Derivatives can be prepare by any variety of
synthetic methods or appropriate adaptations presented in synthetic
or organic chemistry text books, such as those provide in March's
Advanced Organic Chemistry: Reactions, Mechanisms, and Structure,
Wiley, 6th Edition (2007) Michael B. Smith or Domino Reactions in
Organic Synthesis, Wiley (2006) Lutz F. Tietze.
[0083] The term "substituted" refers to a molecule wherein at least
one hydrogen atom is replaced with a substituent. When substituted,
one or more of the groups are "substituents." The molecule may be
multiply substituted. In the case of an oxo substituent (".dbd.O"),
two hydrogen atoms are replaced. Example substituents within this
context may include halogen, hydroxy, alkyl, alkoxy, nitro, cyano,
oxo, carbocyclyl, carbocycloalkyl, heterocarbocyclyl,
heterocarbocycloalkyl, aryl, arylalkyl, heteroaryl,
heteroarylalkyl, --NRaRb, --NRaC(.dbd.O)Rb, --NRaC(.dbd.O)NRaNRb,
--NRaC(.dbd.O)ORb, --NRaSO2Rb, --C(.dbd.O)Ra, --C(.dbd.O)ORa,
--C(.dbd.O)NRaRb, --OC(.dbd.O)NRaRb, --ORa, --SRa, --SORa,
--S(.dbd.O)2Ra, --OS(.dbd.O)2Ra and --S(.dbd.O)2ORa. Ra and Rb in
this context may be the same or different and independently
hydrogen, halogen hydroxy, alkyl, alkoxy, alkyl, amino, alkylamino,
dialkylamino, carbocyclyl, carbocycloalkyl, heterocarbocyclyl,
heterocarbocycloalkyl, aryl, arylalkyl, heteroaryl,
heteroarylalkyl.
[0084] The term "optionally substituted," as used herein, means
that substitution is optional and therefore it is possible for the
designated atom to be unsubstituted.
[0085] As used herein, "subject" refers to any animal, preferably a
human patient, livestock, or domestic pet.
[0086] As used herein, the terms "prevent" and "preventing" include
the prevention of the recurrence, spread or onset. It is not
intended that the present disclosure be limited to complete
prevention. In some embodiments, the onset is delayed, or the
severity is reduced.
[0087] As used herein, the terms "treat" and "treating" are not
limited to the case where the subject (e.g. patient) is cured and
the disease is eradicated. Rather, embodiments of the present
disclosure also contemplate treatment that merely reduces symptoms,
and/or delays disease progression.
[0088] As used herein, the term "calcium phosphate(s)" refers to
minerals containing calcium ions together with orthophosphates,
metaphosphates or pyrophosphates and optionally hydroxide ions.
Tricalcium phosphate is a calcium phosphate with formula
Ca.sub.3(PO.sub.4).sub.2. The common mineral apatite has the basic
formula Ca.sub.5(PO.sub.4).sub.3X, where X is a ion, typically a
halogen or hydroxide ion, or a mixture. Hydroxyapatite refers to
apatite where X is mainly hydroxide ion. In certain embodiments,
the disclosure contemplates calium phosphates that can further
include both silicate (SiO.sub.4.sup.4-) and carbonate
(CO.sub.3.sup.2-) substituted hydroxyapatites, where the
substitution is for one or more of the hydroxy and/or phosphate
groups.
[0089] When used in reference to compound(s) disclosed herein,
"salts" refer to derivatives of the disclosed compound(s) where the
parent compound is modified making acid or base salts thereof.
Examples of salts include, but are not limited to, mineral or
organic acid salts of basic residues such as amines, alkylamines,
or dialkylamines; alkali or organic salts of acidic residues such
as carboxylic acids; and the like.
[0090] As used herein, "alkyl" means a noncyclic straight chain or
branched, unsaturated or saturated hydrocarbon such as those
containing from 1 to 10 carbon atoms. Representative saturated
straight chain alkyls include methyl, ethyl, n-propyl, n-butyl,
n-pentyl, n-hexyl, n-septyl, n-octyl, n-nonyl, and the like; while
saturated branched alkyls include isopropyl, sec-butyl, isobutyl,
tert-butyl, isopentyl, and the like. Unsaturated alkyls contain at
least one double or triple bond between adjacent carbon atoms
(referred to as an "alkenyl" or "alkynyl", respectively).
Representative straight chain and branched alkenyls include
ethylenyl, propylenyl, 1-butenyl, 2-butenyl, isobutylenyl,
1-pentenyl, 2-pentenyl, 3-methyl-1-butenyl, 2-methyl-2-butenyl,
2,3-dimethyl-2-butenyl, and the like; while representative straight
chain and branched alkynyls include acetylenyl, propynyl,
1-butynyl, 2-butynyl, 1-pentynyl, 2-pentynyl, 3-methyl-1-butynyl,
and the like.
[0091] Non-aromatic mono or polycyclic alkyls are referred to
herein as "carbocycles" or "carbocyclyl" groups. Representative
saturated carbocycles include cyclopropyl, cyclobutyl, cyclopentyl,
cyclohexyl, and the like; while unsaturated carbocycles include
cyclopentenyl and cyclohexenyl, and the like.
[0092] "Heterocarbocycles" or heterocarbocyclyl" groups are
carbocycles which contain from 1 to 4 heteroatoms independently
selected from nitrogen, oxygen and sulfur which may be saturated or
unsaturated (but not aromatic), monocyclic or polycyclic, and
wherein the nitrogen and sulfur heteroatoms may be optionally
oxidized, and the nitrogen heteroatom may be optionally
quaternized. Heterocarbocycles include morpholinyl, pyrrolidinonyl,
pyrrolidinyl, piperidinyl, hydantoinyl, valerolactamyl, oxiranyl,
oxetanyl, tetrahydrofuranyl, tetrahydropyranyl,
tetrahydropyridinyl, tetrahydroprimidinyl, tetrahydrothiophenyl,
tetrahydrothiopyranyl, tetrahydropyrimidinyl, tetrahydrothiophenyl,
tetrahydrothiopyranyl, and the like.
[0093] "Aryl" means an aromatic carbocyclic monocyclic or
polycyclic ring such as phenyl or naphthyl. Polycyclic ring systems
may, but are not required to, contain one or more non-aromatic
rings, as long as one of the rings is aromatic.
[0094] As used herein, "heteroaryl" or "heteroaromatic" refers an
aromatic heterocarbocycle having 1 to 4 heteroatoms selected from
nitrogen, oxygen and sulfur, and containing at least 1 carbon atom,
including both mono- and polycyclic ring systems. Polycyclic ring
systems may, but are not required to, contain one or more
non-aromatic rings, as long as one of the rings is aromatic.
Representative heteroaryls are furyl, benzofuranyl, thiophenyl,
benzothiophenyl, pyrrolyl, indolyl, isoindolyl, azaindolyl,
pyridyl, quinolinyl, isoquinolinyl, oxazolyl, isooxazolyl,
benzoxazolyl, pyrazolyl, imidazolyl, benzimidazolyl, thiazolyl,
benzothiazolyl, isothiazolyl, pyridazinyl, pyrimidinyl, pyrazinyl,
triazinyl, cinnolinyl, phthalazinyl, and quinazolinyl. It is
contemplated that the use of the term "heteroaryl" includes
N-alkylated derivatives such as a 1-methylimidazol-5-yl
substituent.
[0095] As used herein, "heterocycle" or "heterocyclyl" refers to
mono- and polycyclic ring systems having 1 to 4 heteroatoms
selected from nitrogen, oxygen and sulfur, and containing at least
1 carbon atom. The mono- and polycyclic ring systems may be
aromatic, non-aromatic or mixtures of aromatic and non-aromatic
rings. Heterocycle includes heterocarbocycles, heteroaryls, and the
like.
[0096] "Alkylthio" refers to an alkyl group as defined above
attached through a sulfur bridge. An example of an alkylthio is
methylthio, (i.e., --S--CH.sub.3).
[0097] "Alkoxy" refers to an alkyl group as defined above attached
through an oxygen bridge. Examples of alkoxy include, but are not
limited to, methoxy, ethoxy, n-propoxy, i-propoxy, n-butoxy,
s-butoxy, t-butoxy, n-pentoxy, and s-pentoxy. Preferred alkoxy
groups are methoxy, ethoxy, n-propoxy, i-propoxy, n-butoxy,
s-butoxy, t-butoxy.
[0098] "Alkylamino" refers an alkyl group as defined above attached
through an amino bridge. An example of an alkylamino is
methylamino, (i.e., --NH--CH.sub.3).
[0099] "Alkanoyl" refers to an alkyl as defined above attached
through a carbonyl bridge (i.e., --(C.dbd.O)alkyl).
[0100] "Alkylsulfonyl" refers to an alkyl as defined above attached
through a sulfonyl bridge (i.e., --S(.dbd.O).sub.2alkyl) such as
mesyl and the like, and "Arylsulfonyl" refers to an aryl attached
through a sulfonyl bridge (i.e., --S(.dbd.O).sub.2aryl).
[0101] "Alkylsulfamoyl" refers to an alkyl as defined above
attached through a sulfamoyl bridge (i.e.,
--S(.dbd.O).sub.2NHalkyl), and an "Arylsulfamoyl" refers to an
alkyl attached through a sulfamoyl bridge (i.e.,
--S(.dbd.O).sub.2NHaryl).
[0102] "Alkylsulfinyl" refers to an alkyl as defined above with the
indicated number of carbon atoms attached through a sulfinyl bridge
(i.e. --S(.dbd.O)alkyl).
[0103] The terms "halogen" and "halo" refer to fluorine, chlorine,
bromine, and iodine.
[0104] The term "bone graft composition" refers to materials that
are substantially physiologically compatible when residing in bone
area, void, or exterior site. In certain embodiments, the bone
graft composition may be a bone graft matrix such as a collagen
sponge or a mixture of polymers and salts.
[0105] As used herein, the term "retinoic acid" refers to the
all-trans compound
(2E,4E,6E,8E)-3,7-dimethyl-9-(2,6,6-trimethylcyclohexen-1-yl)non-
a-2,4,6,8-tetraenoic acid also known as tretinoin.
Compounds
[0106] Compounds derivatives disclosed herein may be used for bone
and cartilage growth and related applications. Derivatives of
certain compounds are further exemplified below.
[0107] In certain embodiments, the disclosure relates to
1-triarylmethyl-1H-imidazole derivatives of formula I:
##STR00001##
or a salt thereof, wherein
[0108] A, E, and G are each the same or different alkyl, alkynyl,
alkenyl, aryl or heteroaryl, wherein A, E, and G are optionally
substituted with one or more, the same or different, R.sup.4;
[0109] R.sup.1, R.sup.2, and R.sup.3 are each the same or different
hydrogen, alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto,
formyl, carboxy, alkanoyl, carbamoyl, alkoxy, alkylthio,
alkylamino, (alkyl).sub.2amino, alkylsulfinyl, alkylsulfonyl,
arylsulfonyl, carbocyclyl, aryl, or heterocyclyl, wherein each
R.sup.1, R.sup.2, and R.sup.3 are optionally substituted with one
or more, the same or different, R.sup.4;
[0110] R.sup.4 is alkyl, halogen, nitro, cyano, hydroxy, amino,
mercapto, formyl, carboxy, alkanoyl, carbamoyl, alkoxy, alkylthio,
alkylamino, (alkyl).sub.2amino, alkylsulfinyl, alkylsulfonyl,
arylsulfonyl, carbocyclyl, aryl, or heterocyclyl, wherein R.sup.4
is optionally substituted with one or more, the same or different,
R.sup.5; and
[0111] R.sup.5 is halogen, nitro, cyano, hydroxy, trifluoromethoxy,
trifluoromethyl, amino, formyl, carboxy, carbamoyl, mercapto,
sulfamoyl, methyl, ethyl, methoxy, ethoxy, acetyl, acetoxy,
methylamino, ethylamino, dimethylamino, diethylamino,
N-methyl-N-ethylamino, acetylamino, N-methylcarbamoyl,
N-ethylcarbamoyl, N,N-dimethylcarbamoyl, N,N-diethylcarbamoyl,
N-methyl-N-ethylcarbamoyl, methylthio, ethylthio, methylsulfinyl,
ethylsulfinyl, mesyl, ethylsulfonyl, methoxycarbonyl,
ethoxycarbonyl, N-methylsulfamoyl, N-ethylsulfamoyl,
N,N-dimethylsulfamoyl, N,N-diethylsulfamoyl,
N-methyl-N-ethylsulfamoyl, carbocyclyl, aryl, or heterocyclyl.
[0112] In certain embodiments, A is alkynyl.
[0113] In certain embodiments, the disclosure relates to
1-triarylmethyl-1H-imidazole derivatives of formula IA:
##STR00002##
[0114] or salts thereof wherein,
[0115] R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.5, R.sup.6,
R.sup.7, R.sup.8, R.sup.9, R.sup.10, R.sup.11, R.sup.12, R.sup.13,
R.sup.14, R.sup.15, R.sup.16, R.sup.17 and R.sup.18 are each the
same or different hydrogen, alkyl, halogen, nitro, cyano, hydroxy,
amino, mercapto, formyl, carboxy, alkanoyl, carbamoyl, alkoxy,
alkylthio, alkylamino, (alkyl).sub.2amino, alkylsulfinyl,
alkylsulfonyl, arylsulfonyl, carbocyclyl, aryl, or heterocyclyl,
wherein each R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.5, R.sup.6,
R.sup.7, R.sup.8, R.sup.9, R.sup.10, R.sup.11, R.sup.12, R.sup.13,
R.sup.14, R.sup.15, R.sup.16, R.sup.17 and R.sup.18 are optionally
substituted with one or more, the same or different, R.sup.19;
[0116] R.sup.19 is alkyl, halogen, nitro, cyano, hydroxy, amino,
mercapto, formyl, carboxy, alkanoyl, carbamoyl, alkoxy, alkylthio,
alkylamino, (alkyl).sub.2amino, alkylsulfinyl, alkylsulfonyl,
arylsulfonyl, carbocyclyl, aryl, or heterocyclyl, wherein R.sup.19
is optionally substituted with one or more, the same or different,
R.sup.20; and
[0117] R.sup.20 is halogen, nitro, cyano, hydroxy,
trifluoromethoxy, trifluoromethyl, amino, formyl, carboxy,
carbamoyl, mercapto, sulfamoyl, methyl, ethyl, methoxy, ethoxy,
acetyl, acetoxy, methylamino, ethylamino, dimethylamino,
diethylamino, N-methyl-N-ethylamino, acetylamino,
N-methylcarbamoyl, N-ethylcarbamoyl, N,N-dimethylcarbamoyl,
N,N-diethylcarbamoyl, N-methyl-N-ethylcarbamoyl, methylthio,
ethylthio, methylsulfinyl, ethylsulfinyl, mesyl, ethylsulfonyl,
methoxycarbonyl, ethoxycarbonyl, N-methylsulfamoyl,
N-ethylsulfamoyl, N,N-dimethylsulfamoyl, N,N-diethylsulfamoyl,
N-methyl-N-ethylsulfamoyl, carbocyclyl, aryl, or heterocyclyl.
[0118] In certain embodiments, R.sup.1, R.sup.2, R.sup.3, R.sup.4,
R.sup.5, R.sup.6, R.sup.7, R.sup.8, R.sup.9, R.sup.10, R.sup.11,
R.sup.12, R.sup.13, R.sup.14, R.sup.15, R.sup.16, and R.sup.17 are
hydrogen and R.sup.18 is a halogen.
[0119] In certain embodiments, the disclosure relates to
biphenyl-diol derivatives of formula II,
##STR00003##
[0120] or salts thereof wherein,
[0121] R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.5, R.sup.6,
R.sup.7, R.sup.8, R.sup.9, and R.sup.10 are each the same or
different hydrogen, alkyl, alkenyl, halogen, nitro, cyano, hydroxy,
amino, mercapto, formyl, carboxy, alkanoyl, carbamoyl, alkoxy,
alkylthio, alkylamino, (alkyl).sub.2amino, alkylsulfinyl,
alkylsulfonyl, arylsulfonyl, carbocyclyl, aryl, or heterocyclyl,
wherein each R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.5, R.sup.6,
R.sup.7, R.sup.8, R.sup.9, and R.sup.10, are optionally substituted
with one or more, the same or different, R.sup.11;
[0122] R.sup.11 is alkyl, alkenyl, halogen, nitro, cyano, hydroxy,
amino, mercapto, formyl, carboxy, carbamoyl, alkanoyl, alkoxy,
alkylthio, alkylamino, (alkyl).sub.2amino, alkylsulfinyl,
alkylsulfonyl, arylsulfonyl, carbocyclyl, aryl, or heterocyclyl,
wherein R.sup.11 is optionally substituted with one or more, the
same or different, R.sup.12; and
[0123] R.sup.12 is halogen, nitro, cyano, hydroxy,
trifluoromethoxy, trifluoromethyl, amino, formyl, carboxy,
carbamoyl, mercapto, sulfamoyl, methyl, ethyl, methoxy, ethoxy,
acetyl, acetoxy, methylamino, ethylamino, dimethylamino,
diethylamino, N-methyl-N-ethylamino, acetylamino,
N-methylcarbamoyl, N-ethylcarbamoyl, N,N-dimethylcarbamoyl,
N,N-diethylcarbamoyl, N-methyl-N-ethylcarbamoyl, methylthio,
ethylthio, methylsulfinyl, ethylsulfinyl, mesyl, ethylsulfonyl,
methoxycarbonyl, ethoxycarbonyl, N-methylsulfamoyl,
N-ethylsulfamoyl, N,N-dimethylsulfamoyl, N,N-diethylsulfamoyl,
N-methyl-N-ethylsulfamoyl, carbocyclyl, aryl, or heterocyclyl.
[0124] In certain embodiments, R.sup.4 and R.sup.9 are alkenyl.
[0125] In certain embodiments, the disclosure relates to macrolide
lactone derivatives of formula III,
##STR00004##
or salts thereof wherein,
[0126] the dotted line represents a single or double bond;
[0127] R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.5, R.sup.6,
R.sup.7, R.sup.8, R.sup.9, R.sup.10, R.sup.11, R.sup.12, R.sup.13,
R.sup.14, R.sup.15, R.sup.16, and R.sup.17 are each the same or
different hydrogen, alkyl, alkenyl, halogen, nitro, cyano, hydroxy,
amino, mercapto, formyl, carboxy, alkanoyl, carbamoyl, alkoxy,
alkylthio, alkylamino, (alkyl).sub.2amino, alkylsulfinyl,
alkylsulfonyl, arylsulfonyl, carbocyclyl, aryl, or heterocyclyl,
wherein each R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.5, R.sup.6,
R.sup.7, R.sup.8, R.sup.9, R.sup.10, R.sup.11, R.sup.12, R.sup.13,
R.sup.14, R.sup.15, R.sup.16, and R.sup.17 are optionally
substituted with one or more, the same or different, R.sup.18;
[0128] R.sup.18 is alkyl, halogen, nitro, cyano, hydroxy, amino,
mercapto, formyl, carboxy, alkanoyl, carbamoyl, alkoxy, alkylthio,
alkylamino, (alkyl).sub.2amino, alkylsulfinyl, alkylsulfonyl,
arylsulfonyl, carbocyclyl, aryl, or heterocyclyl, wherein R.sup.18
is optionally substituted with one or more, the same or different,
R.sup.19; and
[0129] R.sup.19 is halogen, nitro, cyano, hydroxy,
trifluoromethoxy, trifluoromethyl, amino, formyl, carboxy,
carbamoyl, mercapto, sulfamoyl, methyl, ethyl, methoxy, ethoxy,
acetyl, acetoxy, methylamino, ethylamino, dimethylamino,
diethylamino, N-methyl-N-ethylamino, acetylamino,
N-methylcarbamoyl, N-ethylcarbamoyl, N,N-dimethylcarbamoyl,
N,N-diethylcarbamoyl, N-methyl-N-ethylcarbamoyl, methylthio,
ethylthio, methylsulfinyl, ethylsulfinyl, mesyl, ethylsulfonyl,
methoxycarbonyl, ethoxycarbonyl, N-methylsulfamoyl,
N-ethylsulfamoyl, N,N-dimethylsulfamoyl, N,N-diethylsulfamoyl,
N-methyl-N-ethylsulfamoyl, carbocyclyl, aryl, or heterocyclyl.
[0130] In certain embodiments, R.sup.1 is alkyl or alkenyl wherein
R.sup.1 is substituted with a carbocyclyl substituted with one or
more R.sup.18.
[0131] In certain embodiments, R.sup.2, R.sup.3, R.sup.4, R.sup.5,
R.sup.7, R.sup.9, R.sup.10, R.sup.12, and R.sup.13 are each the
same or different alkyl, alkenyl, hydroxy, or alkoxy.
[0132] In certain embodiments, R.sup.6, R.sup.8, R.sup.11,
R.sup.14, R.sup.15, R.sup.16, and R.sup.17 are all hydrogen.
[0133] In certain embodiments, the disclosure relates to macrolide
lactone derivatives of formula IIIA,
##STR00005##
[0134] or salts thereof wherein,
[0135] the dotted lines each individually represent a single or
double bond;
[0136] X is hydroxy, amino, mercapto, or halogen;
[0137] R.sup.2, R.sup.3, R.sup.4, R.sup.5, R.sup.7, R.sup.9,
R.sup.10, R.sup.12, R.sup.13, R.sup.18 and R.sup.19 are each the
same or different hydrogen, alkyl, alkenyl, halogen, nitro, cyano,
hydroxy, amino, mercapto, formyl, carboxy, alkanoyl, carbamoyl,
alkoxy, alkylthio, alkylamino, (alkyl).sub.2amino, alkylsulfinyl,
alkylsulfonyl, arylsulfonyl, carbocyclyl, aryl, or heterocyclyl,
wherein each R.sup.2, R.sup.3, R.sup.4, R.sup.5, R.sup.7, R.sup.9,
R.sup.10, R.sup.12, R.sup.13, R.sup.18 and R.sup.19 are optionally
substituted with one or more, the same or different, R.sup.20;
[0138] R.sup.20 is alkyl, halogen, nitro, cyano, hydroxy, amino,
mercapto, formyl, carboxy, alkanoyl, carbamoyl, alkoxy, alkylthio,
alkylamino, (alkyl).sub.2amino, alkylsulfinyl, alkylsulfonyl,
arylsulfonyl, carbocyclyl, aryl, or heterocyclyl, wherein R.sup.20
is optionally substituted with one or more, the same or different,
R.sup.21; and
[0139] R.sup.21 is halogen, nitro, cyano, hydroxy,
trifluoromethoxy, trifluoromethyl, amino, formyl, carboxy,
carbamoyl, mercapto, sulfamoyl, methyl, ethyl, methoxy, ethoxy,
acetyl, acetoxy, methylamino, ethylamino, dimethylamino,
diethylamino, N-methyl-N-ethylamino, acetylamino,
N-methylcarbamoyl, N-ethylcarbamoyl, N,N-dimethylcarbamoyl,
N,N-diethylcarbamoyl, N-methyl-N-ethylcarbamoyl, methylthio,
ethylthio, methylsulfinyl, ethylsulfinyl, mesyl, ethylsulfonyl,
methoxycarbonyl, ethoxycarbonyl, N-methylsulfamoyl,
N-ethylsulfamoyl, N,N-dimethylsulfamoyl, N,N-diethylsulfamoyl,
N-methyl-N-ethylsulfamoyl, carbocyclyl, aryl, or heterocyclyl.
[0140] In certain embodiments, R.sup.2, R.sup.3, R.sup.4, R.sup.5,
R.sup.7, R.sup.9, R.sup.10, R.sup.12, R.sup.13, R.sup.18 and
R.sup.19 the same or different hydrogen, alkyl, alkenyl, halogen,
hydroxy, or alkoxy.
[0141] In certain embodiments, X is hydroxy or halogen.
[0142] In certain embodiments, R.sup.2, R.sup.5, R.sup.7, and
R.sup.18 are alkyl.
[0143] In certain embodiments, R.sup.3 and R.sup.13 are
hydroxy.
[0144] In certain embodiment, R.sup.4 is alkenyl.
[0145] In certain embodiment, R.sup.9, R.sup.10, and R.sup.19 are
alkoxy.
[0146] In certain embodiments, the disclosure relates to steroid
derivatives. In certain embodiments, the steroid derivatives have
formula IV:
##STR00006##
[0147] or salts thereof wherein,
[0148] the dotted lines represent an optional bond to provide an
absent, single, or double bond
[0149] a) provided that if the dotted ring is aromatic, then the
dotted line between the O and the ring is a single bond and the
dotted line between the ring and R.sup.5 is absent as R.sup.5 is
absent and
[0150] b) provided that if the dotted line between O and the ring
is a double bond, then the dotted line between the O and R.sup.9 is
absent as R.sup.9 is absent;
[0151] R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.5, R.sup.6,
R.sup.7, R.sup.8, R.sup.9, and R.sup.10 are each the same or
different hydrogen, alkyl, alkenyl, alkynyl, halogen, nitro, cyano,
hydroxy, amino, mercapto, formyl, carboxy, alkanoyl, carbamoyl,
alkoxy, alkylthio, alkylamino, (alkyl).sub.2amino, alkylsulfinyl,
alkylsulfonyl, arylsulfonyl, carbocyclyl, aryl, or heterocyclyl,
wherein each R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.5, R.sup.6,
R.sup.7, R.sup.8, R.sup.9, and R.sup.10 are optionally substituted
with one or more, the same or different, R.sup.11;
[0152] R.sup.1 and R.sup.2 may form a carbocyclic or heterocyclic
ring optionally substituted with one or more, the same or
different, R.sup.11.
[0153] R.sup.11 is alkyl, halogen, nitro, cyano, hydroxy, amino,
mercapto, formyl, carboxy, alkanoyl, carbamoyl, alkoxy, alkylthio,
alkylamino, (alkyl).sub.2amino, alkylsulfinyl, alkylsulfonyl,
arylsulfonyl, carbocyclyl, aryl, or heterocyclyl, wherein R.sup.11
is optionally substituted with one or more, the same or different,
R.sup.12; and
[0154] R.sup.12 is halogen, nitro, cyano, hydroxy,
trifluoromethoxy, trifluoromethyl, amino, formyl, carboxy,
carbamoyl, mercapto, sulfamoyl, methyl, ethyl, methoxy, ethoxy,
acetyl, acetoxy, methylamino, ethylamino, dimethylamino,
diethylamino, N-methyl-N-ethylamino, acetylamino,
N-methylcarbamoyl, N-ethylcarbamoyl, N,N-dimethylcarbamoyl,
N,N-diethylcarbamoyl, N-methyl-N-ethylcarbamoyl, methylthio,
ethylthio, methylsulfinyl, ethylsulfinyl, mesyl, ethylsulfonyl,
methoxycarbonyl, ethoxycarbonyl, N-methylsulfamoyl,
N-ethylsulfamoyl, N,N-dimethylsulfamoyl, N,N-diethylsulfamoyl,
N-methyl-N-ethylsulfamoyl, carbocyclyl, aryl, or heterocyclyl.
[0155] In certain embodiments, R.sup.1 and R.sup.2 form a
5-membered lactone ring.
[0156] In certain embodiments, R.sup.2 is hydroxy.
[0157] In certain embodiments, R.sup.10 is hydrogen or mercapto
optionally substituted with acetyl.
[0158] In certain embodiments, the dotted ring is aromatic and
R.sup.9 is hydrogen.
[0159] In certain embodiments, the disclosure relates to
3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenant-
hrene-17-carbothioic acid derivatives of formula IVA
##STR00007##
[0160] or salts thereof wherein,
[0161] R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.5, R.sup.6,
R.sup.7, and R.sup.8 are each the same or different hydrogen,
alkyl, alkenyl, halogen, nitro, cyano, hydroxy, amino, mercapto,
formyl, carboxy, alkanoyl, carbamoyl, alkoxy, alkylthio,
alkylamino, (alkyl).sub.2amino, alkylsulfinyl, alkylsulfonyl,
arylsulfonyl, carbocyclyl, aryl, or heterocyclyl, wherein each
R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.5, R.sup.6, R.sup.7, and
R.sup.8 are optionally substituted with one or more, the same or
different, R.sup.10;
[0162] R.sup.10 is alkyl, halogen, nitro, cyano, hydroxy, amino,
mercapto, formyl, carboxy, alkanoyl, carbamoyl, alkoxy, alkylthio,
alkylamino, (alkyl).sub.2amino, alkylsulfinyl, alkylsulfonyl,
arylsulfonyl, carbocyclyl, aryl, or heterocyclyl, wherein R.sup.10
is optionally substituted with one or more, the same or different,
R.sup.11; and
[0163] R.sup.11 is halogen, nitro, cyano, hydroxy,
trifluoromethoxy, trifluoromethyl, amino, formyl, carboxy,
carbamoyl, mercapto, sulfamoyl, methyl, ethyl, methoxy, ethoxy,
acetyl, acetoxy, methylamino, ethylamino, dimethylamino,
diethylamino, N-methyl-N-ethylamino, acetylamino,
N-methylcarbamoyl, N-ethylcarbamoyl, N,N-dimethylcarbamoyl,
N,N-diethylcarbamoyl, N-methyl-N-ethylcarbamoyl, methylthio,
ethylthio, methylsulfinyl, ethylsulfinyl, mesyl, ethylsulfonyl,
methoxycarbonyl, ethoxycarbonyl, N-methylsulfamoyl,
N-ethylsulfamoyl, N,N-dimethylsulfamoyl, N,N-diethylsulfamoyl,
N-methyl-N-ethylsulfamoyl, carbocyclyl, aryl, or heterocyclyl.
[0164] In certain embodiments, R.sup.1 is an alkyl substituted with
one or more halogens.
[0165] In certain embodiments, R.sup.2 is hydroxy substituted with
an alkanoyl.
[0166] In certain embodiments, R.sup.4 and R.sup.8 are halogen.
[0167] In certain embodiments, R.sup.6 is hydroxy.
[0168] In certain embodiments, the disclosure relates to
3-(4-morpholinophenyl)oxazolidin-2-one derivatives of formula
V,
##STR00008##
or salts thereof wherein
[0169] X is O, S, or NR.sup.17
[0170] R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.5, R.sup.6,
R.sup.7, R.sup.8, R.sup.9, R.sup.10, R.sup.11, R.sup.12, R.sup.13,
R.sup.14, R.sup.15, R.sup.16, and R.sup.17 are each the same or
different hydrogen, alkyl, alkenyl, halogen, nitro, cyano, hydroxy,
amino, mercapto, formyl, carboxy, alkanoyl, carbamoyl, alkoxy,
alkylthio, alkylamino, (alkyl).sub.2amino, alkylsulfinyl,
alkylsulfonyl, arylsulfonyl, carbocyclyl, aryl, or heterocyclyl,
wherein each R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.5, R.sup.6,
R.sup.7, R.sup.8, R.sup.9, R.sup.10, R.sup.11, R.sup.12, R.sup.13,
R.sup.14, R.sup.15, R.sup.16, and R.sup.17 are optionally
substituted with one or more, the same or different, R.sup.18;
[0171] R.sup.18 is alkyl, halogen, nitro, cyano, hydroxy, amino,
mercapto, formyl, carboxy, alkanoyl, carbamoyl, alkoxy, alkylthio,
alkylamino, (alkyl).sub.2amino, alkylsulfinyl, alkylsulfonyl,
arylsulfonyl, carbocyclyl, aryl, or heterocyclyl, wherein R.sup.18
is optionally substituted with one or more, the same or different,
R.sup.19; and
[0172] R.sup.19 is halogen, nitro, cyano, hydroxy,
trifluoromethoxy, trifluoromethyl, amino, formyl, carboxy,
carbamoyl, mercapto, sulfamoyl, methyl, ethyl, methoxy, ethoxy,
acetyl, acetoxy, methylamino, ethylamino, dimethylamino,
diethylamino, N-methyl-N-ethylamino, acetylamino,
N-methylcarbamoyl, N-ethylcarbamoyl, N,N-dimethylcarbamoyl,
N,N-diethylcarbamoyl, N-methyl-N-ethylcarbamoyl, methylthio,
ethylthio, methylsulfinyl, ethylsulfinyl, mesyl, ethylsulfonyl,
methoxycarbonyl, ethoxycarbonyl, N-methylsulfamoyl,
N-ethylsulfamoyl, N,N-dimethylsulfamoyl, N,N-diethylsulfamoyl,
N-methyl-N-ethylsulfamoyl, carbocyclyl, aryl, or heterocyclyl.
[0173] In certain embodiments, X is O.
[0174] In certain embodiments, R.sup.1 is alkyl substituted with
R.sup.18.
[0175] In certain embodiments R.sup.2, R.sup.3, R.sup.4, R.sup.5,
R.sup.6, R.sup.7, R.sup.8, R.sup.9, R.sup.10, R.sup.11, R.sup.12,
R.sup.13, R.sup.14, R.sup.15, and R.sup.16 are all hydrogen.
[0176] In certain embodiments, the disclosure relates to
3-(4-morpholinophenyl)oxazolidin-2-one derivatives of formula
VI,
##STR00009##
[0177] or salts thereof wherein
[0178] R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.5, R.sup.6,
R.sup.7, R.sup.8, R.sup.9, R.sup.10, R.sup.11, R.sup.12, R.sup.13,
R.sup.14, R.sup.15, R.sup.16, R.sup.17, R.sup.18, R.sup.19, and
R.sup.20 are each the same or different hydrogen, alkyl, halogen,
nitro, cyano, hydroxy, amino, mercapto, formyl, carboxy, alkanoyl,
carbamoyl, alkoxy, alkylthio, alkylamino, (alkyl).sub.2amino,
alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl, aryl, or
heterocyclyl, wherein each R.sup.1, R.sup.2, R.sup.3, R.sup.4,
R.sup.5, R.sup.6, R.sup.7, R.sup.8, R.sup.9, R.sup.10, R.sup.11,
R.sup.12, R.sup.13, R.sup.14, R.sup.15, R.sup.16, R.sup.17,
R.sup.18, R.sup.19, and R.sup.20 are optionally substituted with
one or more, the same or different, R.sup.21;
[0179] R.sup.21 is alkyl, halogen, nitro, cyano, hydroxy, amino,
mercapto, formyl, carboxy, alkanoyl, carbamoyl, alkoxy, alkylthio,
alkylamino, (alkyl).sub.2amino, alkylsulfinyl, alkylsulfonyl,
arylsulfonyl, carbocyclyl, aryl, or heterocyclyl, wherein R.sup.21
is optionally substituted with one or more, the same or different,
R.sup.22; and
[0180] R.sup.22 is halogen, nitro, cyano, hydroxy,
trifluoromethoxy, trifluoromethyl, amino, formyl, carboxy,
carbamoyl, mercapto, sulfamoyl, methyl, ethyl, methoxy, ethoxy,
acetyl, acetoxy, methylamino, ethylamino, dimethylamino,
diethylamino, N-methyl-N-ethylamino, acetylamino,
N-methylcarbamoyl, N-ethylcarbamoyl, N,N-dimethylcarbamoyl,
N,N-diethylcarbamoyl, N-methyl-N-ethylcarbamoyl, methylthio,
ethylthio, methylsulfinyl, ethylsulfinyl, mesyl, ethylsulfonyl,
methoxycarbonyl, ethoxycarbonyl, N-methylsulfamoyl,
N-ethylsulfamoyl, N,N-dimethylsulfamoyl, N,N-diethylsulfamoyl,
N-methyl-N-ethylsulfamoyl, carbocyclyl, aryl, or heterocyclyl.
[0181] In certain embodiments, R.sup.1, R.sup.2, and R.sup.8 are
each the same or different alkyl.
[0182] In certain embodiments, R.sup.14 is carboxy.
[0183] In certain embodiments R.sup.3, R.sup.4, R.sup.5, R.sup.6,
R.sup.7, R.sup.8, R.sup.9, R.sup.10, R.sup.11, R.sup.12, R.sup.13,
R.sup.14, R.sup.15, R.sup.16, R.sup.17, R.sup.18, R.sup.19, and
R.sup.20 are hydrogen.
[0184] In certain embodiments, the disclosure relates to rentinol
derivatives of formula VII,
##STR00010##
[0185] or salts thereof wherein
[0186] the dotted lines represent an optional bond to provide an
absent, single, or double bond provided that if the dotted ring is
aromatic, then the dotted line between R.sup.2 and the ring is
absent as R.sup.2 is absent;
[0187] R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.5, R.sup.6,
R.sup.7, R.sup.8, R.sup.9, R.sup.10, R.sup.11, R.sup.12, R.sup.13,
and R.sup.14 are each the same or different hydrogen, alkyl,
halogen, nitro, cyano, hydroxy, amino, mercapto, formyl, carboxy,
alkanoyl, carbamoyl, alkoxy, alkylthio, alkylamino,
(alkyl).sub.2amino, alkylsulfinyl, alkylsulfonyl, arylsulfonyl,
carbocyclyl, aryl, or heterocyclyl, wherein each R.sup.1, R.sup.2,
R.sup.3, R.sup.4, R.sup.5, R.sup.6, R.sup.7, R.sup.8, R.sup.9,
R.sup.10, R.sup.11, R.sup.12, R.sup.13, and R.sup.14 are optionally
substituted with one or more, the same or different, R.sup.15;
[0188] R.sup.15 is alkyl, halogen, nitro, cyano, hydroxy, amino,
mercapto, formyl, carboxy, alkanoyl, carbamoyl, alkoxy, alkylthio,
alkylamino, (alkyl).sub.2amino, alkylsulfinyl, alkylsulfonyl,
arylsulfonyl, carbocyclyl, aryl, or heterocyclyl, wherein R.sup.15
is optionally substituted with one or more, the same or different,
R.sup.16; and
[0189] R.sup.16 is halogen, nitro, cyano, hydroxy,
trifluoromethoxy, trifluoromethyl, amino, formyl, carboxy,
carbamoyl, mercapto, sulfamoyl, methyl, ethyl, methoxy, ethoxy,
acetyl, acetoxy, methylamino, ethylamino, dimethylamino,
diethylamino, N-methyl-N-ethylamino, acetylamino,
N-methylcarbamoyl, N-ethylcarbamoyl, N,N-dimethylcarbamoyl,
N,N-diethylcarbamoyl, N-methyl-N-ethylcarbamoyl, methylthio,
ethylthio, methylsulfinyl, ethylsulfinyl, mesyl, ethylsulfonyl,
methoxycarbonyl, ethoxycarbonyl, N-methylsulfamoyl,
N-ethylsulfamoyl, N,N-dimethylsulfamoyl, N,N-diethylsulfamoyl,
N-methyl-N-ethylsulfamoyl, carbocyclyl, aryl, or heterocyclyl.
[0190] In certain embodiments, R.sup.1, R.sup.2, R.sup.6, R.sup.11
and R.sup.13 are alkyl.
[0191] In certain embodiments, R.sup.3, R.sup.4, R.sup.5, R.sup.7,
R.sup.8, R.sup.9, R.sup.12, and R.sup.14 are hydrogen.
[0192] In certain embodiments, the dotted ring is aromatic and
R.sup.1, R.sup.3, R.sup.4, and R.sup.6 are alkyl or alkoxy.
[0193] In certain embodiments R.sup.10 is hydroxy, formyl, or
carboxy optionally substituted with R.sup.15.
[0194] In certain embodiments, the disclosure relates to rentinol
derivatives of formula VIIA,
##STR00011##
[0195] or salts thereof wherein
[0196] R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.5, R.sup.6,
R.sup.7, R.sup.8, R.sup.9, R.sup.10, R.sup.11, R.sup.12, R.sup.13,
and R.sup.14 are each the same or different hydrogen, alkyl,
halogen, nitro, cyano, hydroxy, amino, mercapto, formyl, carboxy,
alkanoyl, carbamoyl, alkoxy, alkylthio, alkylamino,
(alkyl).sub.2amino, alkylsulfinyl, alkylsulfonyl, arylsulfonyl,
carbocyclyl, aryl, or heterocyclyl, wherein each R.sup.1, R.sup.2,
R.sup.3, R.sup.4, R.sup.5, R.sup.6, R.sup.7, R.sup.8, R.sup.9,
R.sup.10, R.sup.11, R.sup.12, R.sup.13, and R.sup.14 are optionally
substituted with one or more, the same or different, R.sup.15;
[0197] R.sup.15 is alkyl, halogen, nitro, cyano, hydroxy, amino,
mercapto, formyl, carboxy, alkanoyl, carbamoyl, alkoxy, alkylthio,
alkylamino, (alkyl).sub.2amino, alkylsulfinyl, alkylsulfonyl,
arylsulfonyl, carbocyclyl, aryl, or heterocyclyl, wherein R.sup.15
is optionally substituted with one or more, the same or different,
R.sup.16; and
[0198] R.sup.16 is halogen, nitro, cyano, hydroxy,
trifluoromethoxy, trifluoromethyl, amino, formyl, carboxy,
carbamoyl, mercapto, sulfamoyl, methyl, ethyl, methoxy, ethoxy,
acetyl, acetoxy, methylamino, ethylamino, dimethylamino,
diethylamino, N-methyl-N-ethylamino, acetylamino,
N-methylcarbamoyl, N-ethylcarbamoyl, N,N-dimethylcarbamoyl,
N,N-diethylcarbamoyl, N-methyl-N-ethylcarbamoyl, methylthio,
ethylthio, methylsulfinyl, ethylsulfinyl, mesyl, ethylsulfonyl,
methoxycarbonyl, ethoxycarbonyl, N-methylsulfamoyl,
N-ethylsulfamoyl, N,N-dimethylsulfamoyl, N,N-diethylsulfamoyl,
N-methyl-N-ethylsulfamoyl, carbocyclyl, aryl, or heterocyclyl.
[0199] In certain embodiments, R.sup.1, R.sup.2, R.sup.6, R.sup.11,
and R.sup.13 are alkyl.
[0200] In certain embodiments, R.sup.3, R.sup.4, R.sup.5, R.sup.7,
R.sup.8, R.sup.9, R.sup.12, and R.sup.14 are hydrogen.
[0201] In certain embodiments, R.sup.10 is hydroxy, formyl, or
carboxy optionally substituted with R.sup.15.
[0202] In certain embodiments, the disclosure relates to rentinol
derivatives of formula VIIB,
##STR00012##
[0203] or salts thereof wherein
[0204] R.sup.1, R.sup.3, R.sup.4, R.sup.5, R.sup.6, R.sup.7,
R.sup.8, R.sup.9, R.sup.10, R.sup.11, R.sup.12, R.sup.13, and
R.sup.14 are each the same or different hydrogen, alkyl, halogen,
nitro, cyano, hydroxy, amino, mercapto, formyl, carboxy, alkanoyl,
carbamoyl, alkoxy, alkylthio, alkylamino, (alkyl).sub.2amino,
alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl, aryl, or
heterocyclyl, wherein each R.sup.1, R.sup.3, R.sup.4, R.sup.5,
R.sup.6, R.sup.7, R.sup.8, R.sup.9, R.sup.10, R.sup.11, R.sup.12,
R.sup.13, and R.sup.14 are optionally substituted with one or more,
the same or different, R.sup.15;
[0205] R.sup.15 is alkyl, halogen, nitro, cyano, hydroxy, amino,
mercapto, formyl, carboxy, alkanoyl, carbamoyl, alkoxy, alkylthio,
alkylamino, (alkyl).sub.2amino, alkylsulfinyl, alkylsulfonyl,
arylsulfonyl, carbocyclyl, aryl, or heterocyclyl, wherein R.sup.15
is optionally substituted with one or more, the same or different,
R.sup.16; and
[0206] R.sup.16 is halogen, nitro, cyano, hydroxy,
trifluoromethoxy, trifluoromethyl, amino, formyl, carboxy,
carbamoyl, mercapto, sulfamoyl, methyl, ethyl, methoxy, ethoxy,
acetyl, acetoxy, methylamino, ethylamino, dimethylamino,
diethylamino, N-methyl-N-ethylamino, acetylamino,
N-methylcarbamoyl, N-ethylcarbamoyl, N,N-dimethylcarbamoyl,
N,N-diethylcarbamoyl, N-methyl-N-ethylcarbamoyl, methylthio,
ethylthio, methylsulfinyl, ethylsulfinyl, mesyl, ethylsulfonyl,
methoxycarbonyl, ethoxycarbonyl, N-methylsulfamoyl,
N-ethylsulfamoyl, N,N-dimethylsulfamoyl, N,N-diethylsulfamoyl,
N-methyl-N-ethylsulfamoyl, carbocyclyl, aryl, or heterocyclyl.
[0207] In certain embodiments, R.sup.1, R.sup.3, R.sup.6, R.sup.11,
and R.sup.13 are alkyl.
[0208] In certain embodiments, R.sup.4 is alkoxy.
[0209] In certain embodiments, R.sup.5, R.sup.7, R.sup.8, R.sup.9,
R.sup.12, and R.sup.14 are hydrogen.
[0210] In certain embodiments, R.sup.10 is hydroxy, formyl, or
carboxy optionally substituted with R.sup.15.
[0211] In certain embodiments, the disclosure relates to
4-(4-(dialkylamino)phenyl)butanoic acid derivatives of formula
VIII,
##STR00013##
[0212] or salts thereof wherein
[0213] n is 1, 2, or 3;
[0214] m is 1, 2, or 3;
[0215] R.sup.1 and R.sup.2 are each the same or different hydrogen,
alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto, formyl,
carboxy, alkanoyl, carbamoyl, alkoxy, alkylthio, alkylamino,
(alkyl).sub.2amino, alkylsulfinyl, alkylsulfonyl, arylsulfonyl,
carbocyclyl, aryl, or heterocyclyl, wherein each R.sup.1 and
R.sup.2 are optionally substituted with one or more, the same or
different, R.sup.14;
[0216] R.sup.3, R.sup.4, R.sup.5, R.sup.6, R.sup.7, R.sup.8,
R.sup.9, R.sup.10, R.sup.11, R.sup.12, and R.sup.13 are each the
same or different hydrogen, alkyl, halogen, nitro, cyano, hydroxy,
amino, mercapto, formyl, carboxy, alkanoyl, carbamoyl, alkoxy,
alkylthio, alkylamino, (alkyl).sub.2amino, alkylsulfinyl,
alkylsulfonyl, arylsulfonyl, carbocyclyl, aryl, or heterocyclyl,
wherein each R.sup.3, R.sup.4, R.sup.5, R.sup.6, R.sup.7, R.sup.8,
R.sup.9, R.sup.10, R.sup.11, R.sup.12, and R.sup.13 are optionally
substituted with one or more, the same or different, R.sup.14;
[0217] R.sup.14 is alkyl, halogen, nitro, cyano, hydroxy, amino,
mercapto, formyl, carboxy, alkanoyl, carbamoyl, alkoxy, alkylthio,
alkylamino, (alkyl).sub.2amino, alkylsulfinyl, alkylsulfonyl,
arylsulfonyl, carbocyclyl, aryl, or heterocyclyl, wherein R.sup.14
is optionally substituted with one or more, the same or different,
R.sup.15; and
[0218] R.sup.15 is halogen, nitro, cyano, hydroxy,
trifluoromethoxy, trifluoromethyl, amino, formyl, carboxy,
carbamoyl, mercapto, sulfamoyl, methyl, ethyl, methoxy, ethoxy,
acetyl, acetoxy, methylamino, ethylamino, dimethylamino,
diethylamino, N-methyl-N-ethylamino, acetylamino,
N-methylcarbamoyl, N-ethylcarbamoyl, N,N-dimethylcarbamoyl,
N,N-diethylcarbamoyl, N-methyl-N-ethylcarbamoyl, methylthio,
ethylthio, methylsulfinyl, ethylsulfinyl, mesyl, ethylsulfonyl,
methoxycarbonyl, ethoxycarbonyl, N-methylsulfamoyl,
N-ethylsulfamoyl, N,N-dimethylsulfamoyl, N,N-diethylsulfamoyl,
N-methyl-N-ethylsulfamoyl, carbocyclyl, aryl, or heterocyclyl.
[0219] In certain embodiments, R.sup.1 and R.sup.2 are a
halogen.
[0220] In certain embodiments, R.sup.3, R.sup.4, R.sup.5, and
R.sup.6 are hydrogen.
[0221] In certain embodiments, R.sup.7, R.sup.8, R.sup.9, R.sup.10,
R.sup.11, and R.sup.12 are hydrogen.
[0222] In certain embodiments, R.sup.13 is hydrogen or alkyl.
[0223] In certain embodiments, the disclosure relates to macrolide
lactone derivatives of formula IX,
##STR00014##
[0224] or salts thereof wherein,
[0225] X is hydroxy, amino, mercapto, halogen, or
--O(CH.sub.2).sub.nOH wherein X is optionally substituted with one
or more, the same or different, R.sup.20;
[0226] n is 2, 3, or 4;
[0227] R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.5, R.sup.6,
R.sup.7, R.sup.8, R.sup.9, R.sup.10, R.sup.11, R.sup.12, R.sup.13,
R.sup.18, and R.sup.19 are each the same or different hydrogen,
alkyl, alkenyl, halogen, nitro, cyano, hydroxy, amino, mercapto,
formyl, carboxy, alkanoyl, carbamoyl, alkoxy, alkylthio,
alkylamino, (alkyl).sub.2amino, alkylsulfinyl, alkylsulfonyl,
arylsulfonyl, carbocyclyl, aryl, or heterocyclyl, wherein each
R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.5, R.sup.6, R.sup.7,
R.sup.8, R.sup.9, R.sup.10, R.sup.11, R.sup.12, R.sup.13, R.sup.18,
and R.sup.19 are optionally substituted with one or more, the same
or different, R.sup.20;
[0228] R.sup.20 is alkyl, halogen, nitro, cyano, hydroxy, amino,
mercapto, formyl, carboxy, alkanoyl, carbamoyl, alkoxy, alkylthio,
alkylamino, (alkyl).sub.2amino, alkylsulfinyl, alkylsulfonyl,
arylsulfonyl, carbocyclyl, aryl, or heterocyclyl, wherein R.sup.20
is optionally substituted with one or more, the same or different,
R.sup.21; and
[0229] R.sup.21 is halogen, nitro, cyano, hydroxy,
trifluoromethoxy, trifluoromethyl, amino, formyl, carboxy,
carbamoyl, mercapto, sulfamoyl, methyl, ethyl, methoxy, ethoxy,
acetyl, acetoxy, methylamino, ethylamino, dimethylamino,
diethylamino, N-methyl-N-ethylamino, acetylamino,
N-methylcarbamoyl, N-ethylcarbamoyl, N,N-dimethylcarbamoyl,
N,N-diethylcarbamoyl, N-methyl-N-ethylcarbamoyl, methylthio,
ethylthio, methylsulfinyl, ethylsulfinyl, mesyl, ethylsulfonyl,
methoxycarbonyl, ethoxycarbonyl, N-methylsulfamoyl,
N-ethylsulfamoyl, N,N-dimethylsulfamoyl, N,N-diethylsulfamoyl,
N-methyl-N-ethylsulfamoyl, carbocyclyl, aryl, or heterocyclyl.
[0230] In certain embodiments R.sup.1, R.sup.2, R.sup.3, R.sup.4,
R.sup.5, R.sup.6, R.sup.7, R.sup.8, R.sup.9, R.sup.10, R.sup.12,
R.sup.13, R.sup.18, and R.sup.19 are each the same or different
hydrogen, alkyl, halogen, hydroxy, or alkoxy.
[0231] In certain embodiments, X is hydroxy or
--O(CH.sub.2).sub.nOH.
[0232] In certain embodiments, n is 2.
[0233] In certain embodiments, R.sup.1, R.sup.2, R.sup.5, R.sup.6,
R.sup.7, R.sup.12, and R.sup.18 are alkyl.
[0234] In certain embodiments, R.sup.3 and R.sup.13 are
hydroxy.
[0235] In certain embodiment, R.sup.4, R.sup.8, and R.sup.19 are
alkoxy.
[0236] In certain embodiments, the podophyllotoxin derivative is a
compound of formula X,
##STR00015##
[0237] or salts thereof wherein,
[0238] R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.5, R.sup.6,
R.sup.7, R.sup.8, R.sup.9, and R.sup.10 are each the same or
different hydrogen, alkyl, alkenyl, halogen, nitro, cyano, hydroxy,
amino, mercapto, formyl, carboxy, alkanoyl, carbamoyl, alkoxy,
alkylthio, alkylamino, (alkyl).sub.2amino, alkylsulfinyl,
alkylsulfonyl, arylsulfonyl, carbocyclyl, aryl, or heterocyclyl,
wherein each R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.5, R.sup.6,
R.sup.7, R.sup.8, R.sup.9, and R.sup.10 are optionally substituted
with one or more, the same or different, R.sup.11;
[0239] R.sup.11 is alkyl, halogen, nitro, cyano, hydroxy, amino,
mercapto, formyl, carboxy, alkanoyl, carbamoyl, alkoxy, alkylthio,
alkylamino, (alkyl).sub.2amino, alkylsulfinyl, alkylsulfonyl,
arylsulfonyl, carbocyclyl, aryl, or heterocyclyl, wherein R.sup.11
is optionally substituted with one or more, the same or different,
R.sup.12; and
[0240] R.sup.12 is halogen, nitro, cyano, hydroxy,
trifluoromethoxy, trifluoromethyl, amino, formyl, carboxy,
carbamoyl, mercapto, sulfamoyl, methyl, ethyl, methoxy, ethoxy,
acetyl, acetoxy, methylamino, ethylamino, dimethylamino,
diethylamino, N-methyl-N-ethylamino, acetylamino,
N-methylcarbamoyl, N-ethylcarbamoyl, N,N-dimethylcarbamoyl,
N,N-diethylcarbamoyl, N-methyl-N-ethylcarbamoyl, methylthio,
ethylthio, methylsulfinyl, ethylsulfinyl, mesyl, ethylsulfonyl,
methoxycarbonyl, ethoxycarbonyl, N-methylsulfamoyl,
N-ethylsulfamoyl, N,N-dimethylsulfamoyl, N,N-diethylsulfamoyl,
N-methyl-N-ethylsulfamoyl, carbocyclyl, aryl, or heterocyclyl.
[0241] In certain embodiments, R.sup.1, R.sup.5, R.sup.8, R.sup.9,
and R.sup.10 are hydrogen.
[0242] In certain embodiments, R.sup.2, R.sup.3, R.sup.4, R.sup.5,
R.sup.6, and R.sup.7 are each are each the same or different
hydroxy or alkoxy.
[0243] In certain embodiments, the aziridine is derivative is a
compound of formula XI,
##STR00016##
[0244] or salts thereof wherein,
[0245] R.sup.1, R.sup.2, R.sup.3, R.sup.4, and R.sup.5 are each the
same or different hydrogen, alkyl, alkenyl, halogen, nitro, cyano,
hydroxy, amino, mercapto, formyl, carboxy, alkanoyl, carbamoyl,
alkoxy, alkylthio, alkylamino, (alkyl).sub.2amino, alkylsulfinyl,
alkylsulfonyl, arylsulfonyl, carbocyclyl, aryl, or heterocyclyl,
wherein each R.sup.1, R.sup.2, R.sup.3, R.sup.4, and R.sup.5 are
optionally substituted with one or more, the same or different,
R.sup.6;
[0246] R.sup.6 is alkyl, halogen, nitro, cyano, hydroxy, amino,
mercapto, formyl, carboxy, alkanoyl, carbamoyl, alkoxy, alkylthio,
alkylamino, (alkyl).sub.2amino, alkylsulfinyl, alkylsulfonyl,
arylsulfonyl, carbocyclyl, aryl, or heterocyclyl, wherein R.sup.6
is optionally substituted with one or more, the same or different,
R.sup.7; and
[0247] R.sup.7 is halogen, nitro, cyano, hydroxy, trifluoromethoxy,
trifluoromethyl, amino, formyl, carboxy, carbamoyl, mercapto,
sulfamoyl, methyl, ethyl, methoxy, ethoxy, acetyl, acetoxy,
methylamino, ethylamino, dimethylamino, diethylamino,
N-methyl-N-ethylamino, acetylamino, N-methylcarbamoyl,
N-ethylcarbamoyl, N,N-dimethylcarbamoyl, N,N-diethylcarbamoyl,
N-methyl-N-ethylcarbamoyl, methylthio, ethylthio, methylsulfinyl,
ethylsulfinyl, mesyl, ethylsulfonyl, methoxycarbonyl,
ethoxycarbonyl, N-methylsulfamoyl, N-ethylsulfamoyl,
N,N-dimethylsulfamoyl, N,N-diethylsulfamoyl,
N-methyl-N-ethylsulfamoyl, carbocyclyl, aryl, or heterocyclyl.
[0248] In certain embodiments, R.sup.1 is alkyl.
[0249] In certain embodiments, R.sup.2 is amino.
[0250] In certain embodiments, R.sup.3 is alkyl substituted with a
carbamoyl group.
[0251] In certain embodiments, R.sup.4 is alkoxy.
[0252] In certain embodiments, the nucleoside derivative is a
compound of formula XII,
##STR00017##
[0253] or salts thereof wherein,
[0254] X is N or CR.sup.6;
[0255] R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.5, and R.sup.6 are
each the same or different hydrogen, alkyl, alkenyl, halogen,
nitro, cyano, hydroxy, amino, mercapto, formyl, carboxy, alkanoyl,
carbamoyl, alkoxy, alkylthio, alkylamino, (alkyl).sub.2amino,
alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl, aryl, or
heterocyclyl, wherein each R.sup.1, R.sup.2, R.sup.3, R.sup.4,
R.sup.5, and R.sup.6 are optionally substituted with one or more,
the same or different, R.sup.7;
[0256] R.sup.7 is alkyl, halogen, nitro, cyano, hydroxy, amino,
mercapto, formyl, carboxy, alkanoyl, carbamoyl, alkoxy, alkylthio,
alkylamino, (alkyl).sub.2amino, alkylsulfinyl, alkylsulfonyl,
arylsulfonyl, carbocyclyl, aryl, or heterocyclyl, wherein R.sup.7
is optionally substituted with one or more, the same or different,
R.sup.8; and
[0257] R.sup.8 is halogen, nitro, cyano, hydroxy, trifluoromethoxy,
trifluoromethyl, amino, formyl, carboxy, carbamoyl, mercapto,
sulfamoyl, methyl, ethyl, methoxy, ethoxy, acetyl, acetoxy,
methylamino, ethylamino, dimethylamino, diethylamino,
N-methyl-N-ethylamino, acetylamino, N-methylcarbamoyl,
N-ethylcarbamoyl, N,N-dimethylcarbamoyl, N,N-diethylcarbamoyl,
N-methyl-N-ethylcarbamoyl, methylthio, ethylthio, methylsulfinyl,
ethylsulfinyl, mesyl, ethylsulfonyl, methoxycarbonyl,
ethoxycarbonyl, N-methylsulfamoyl, N-ethylsulfamoyl,
N,N-dimethylsulfamoyl, N,N-diethylsulfamoyl,
N-methyl-N-ethylsulfamoyl, carbocyclyl, aryl, or heterocyclyl.
[0258] In certain embodiments, R.sup.1 is hydroxymethyl.
[0259] In certain embodiments, R.sup.2 is hydroxy.
[0260] In certain embodiments, R.sup.3 is hydrogen or hydroxy.
[0261] In certain embodiments, R.sup.4 is hydrogen.
[0262] In certain embodiments, R.sup.5 is hydrogen or amino.
[0263] In certain embodiments, X is CH or N.
[0264] In certain embodiments, the vinca alkaloid derivative is a
compound of formula XIII,
##STR00018##
[0265] or salts thereof wherein,
[0266] n is 1 or 2;
[0267] R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.5, R.sup.6,
R.sup.7, R.sup.8, R.sup.9, R.sup.10, R.sup.11, R.sup.12, R.sup.13,
R.sup.14, R.sup.15, R.sup.16, R.sup.17, R.sup.18, R.sup.19,
R.sup.20, R.sup.21, R.sup.22, R.sup.23, R.sup.24, and R.sup.25 are
each the same or different hydrogen, alkyl, alkenyl, halogen,
nitro, cyano, hydroxy, amino, mercapto, formyl, carboxy, alkanoyl,
carbamoyl, alkoxy, alkylthio, alkylamino, (alkyl).sub.2amino,
alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl, aryl, or
heterocyclyl, wherein each R.sup.1, R.sup.2, R.sup.3, R.sup.4,
R.sup.5, R.sup.6, R.sup.7, R.sup.8, R.sup.9, R.sup.10, R.sup.11,
R.sup.12, R.sup.13, R.sup.14, R.sup.15, R.sup.16, R.sup.17,
R.sup.18, R.sup.19, R.sup.20, R.sup.21, R.sup.22, R.sup.23,
R.sup.24, and R.sup.25 are optionally substituted with one or more,
the same or different, R.sup.26;
[0268] R.sup.26 is alkyl, halogen, nitro, cyano, hydroxy, amino,
mercapto, formyl, carboxy, alkanoyl, carbamoyl, alkoxy, alkylthio,
alkylamino, (alkyl).sub.2amino, alkylsulfinyl, alkylsulfonyl,
arylsulfonyl, carbocyclyl, aryl, or heterocyclyl, wherein R.sup.26
is optionally substituted with one or more, the same or different,
R.sup.27; and
[0269] R.sup.27 is halogen, nitro, cyano, hydroxy,
trifluoromethoxy, trifluoromethyl, amino, formyl, carboxy,
carbamoyl, mercapto, sulfamoyl, methyl, ethyl, methoxy, ethoxy,
acetyl, acetoxy, methylamino, ethylamino, dimethylamino,
diethylamino, N-methyl-N-ethylamino, acetylamino,
N-methylcarbamoyl, N-ethylcarbamoyl, N,N-dimethylcarbamoyl,
N,N-diethylcarbamoyl, N-methyl-N-ethylcarbamoyl, methylthio,
ethylthio, methylsulfinyl, ethylsulfinyl, mesyl, ethylsulfonyl,
methoxycarbonyl, ethoxycarbonyl, N-methylsulfamoyl,
N-ethylsulfamoyl, N,N-dimethylsulfamoyl, N,N-diethylsulfamoyl,
N-methyl-N-ethylsulfamoyl, carbocyclyl, aryl, or heterocyclyl.
[0270] In certain embodiments, R.sup.4 is alkyl.
[0271] In certain embodiments, R.sup.5 is hydroxy substituted with
acetyl.
[0272] In certain embodiments, R.sup.7 is hydroxy.
[0273] In certain embodiments, R.sup.8 is carboxy substituted with
alkyl.
[0274] In certain embodiments, R.sup.10 is alkyl or formyl.
[0275] In certain embodiments, R.sup.12 is alkoxy.
[0276] In certain embodiments, R.sup.13 is carboxy substituted with
alkyl.
[0277] In certain embodiments, R.sup.16 and R.sup.18 form a double
bond.
[0278] In certain embodiments, R.sup.18 is hydroxy.
[0279] In certain embodiments, R.sup.19 is alkyl.
[0280] In certain embodiments, R.sup.1, R.sup.2, R.sup.3, R.sup.6,
R.sup.9, R.sup.11, R.sup.14, R.sup.15, R.sup.16, R.sup.17,
R.sup.20, R.sup.21, R.sup.22, R.sup.23, R.sup.24, and R.sup.25 are
each hydrogen.
[0281] In certain embodiments, the anthracene-9,10-dione
derivatives is a compound of formula XIV,
##STR00019##
[0282] or salts thereof wherein,
[0283] R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.5, R.sup.6,
R.sup.7, and R.sup.8 are each the same or different hydrogen,
alkyl, alkenyl, halogen, nitro, cyano, hydroxy, amino, mercapto,
formyl, carboxy, alkanoyl, carbamoyl, alkoxy, alkylthio,
alkylamino, (alkyl).sub.2amino, alkylsulfinyl, alkylsulfonyl,
arylsulfonyl, carbocyclyl, aryl, or heterocyclyl, wherein each
R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.5, R.sup.6, R.sup.7, and
R.sup.8 are optionally substituted with one or more, the same or
different, R.sup.9;
[0284] R.sup.9 is alkyl, halogen, nitro, cyano, hydroxy, amino,
mercapto, formyl, carboxy, alkanoyl, carbamoyl, alkoxy, alkylthio,
alkylamino, (alkyl).sub.2amino, alkylsulfinyl, alkylsulfonyl,
arylsulfonyl, carbocyclyl, aryl, or heterocyclyl, wherein R.sup.9
is optionally substituted with one or more, the same or different,
R.sup.10; and
[0285] R.sup.10 is halogen, nitro, cyano, hydroxy,
trifluoromethoxy, trifluoromethyl, amino, formyl, carboxy,
carbamoyl, mercapto, sulfamoyl, methyl, ethyl, methoxy, ethoxy,
acetyl, acetoxy, methylamino, ethylamino, dimethylamino,
diethylamino, N-methyl-N-ethylamino, acetylamino,
N-methylcarbamoyl, N-ethylcarbamoyl, N,N-dimethylcarbamoyl,
N,N-diethylcarbamoyl, N-methyl-N-ethylcarbamoyl, methylthio,
ethylthio, methylsulfinyl, ethylsulfinyl, mesyl, ethylsulfonyl,
methoxycarbonyl, ethoxycarbonyl, N-methylsulfamoyl,
N-ethylsulfamoyl, N,N-dimethylsulfamoyl, N,N-diethylsulfamoyl,
N-methyl-N-ethylsulfamoyl, carbocyclyl, aryl, or heterocyclyl.
[0286] In certain embodiments, the anthracycline derivatives is a
compound of formula XV,
##STR00020##
or salts thereof wherein,
[0287] R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.5, R.sup.6,
R.sup.7, R.sup.8, R.sup.9, R.sup.10, R.sup.11, R.sup.12, R.sup.13,
and R.sup.14 are each the same or different hydrogen, alkyl,
alkenyl, halogen, nitro, cyano, hydroxy, amino, mercapto, formyl,
carboxy, alkanoyl, carbamoyl, alkoxy, alkylthio, alkylamino,
(alkyl).sub.2amino, alkylsulfinyl, alkylsulfonyl, arylsulfonyl,
carbocyclyl, aryl, or heterocyclyl, wherein each R.sup.1, R.sup.2,
R.sup.3, R.sup.4, R.sup.5, R.sup.6, R.sup.7, R.sup.8, R.sup.9,
R.sup.10, R.sup.11, R.sup.12, R.sup.13, and R.sup.14 are optionally
substituted with one or more, the same or different, R.sup.15;
[0288] R.sup.15 is alkyl, halogen, nitro, cyano, hydroxy, amino,
mercapto, formyl, carboxy, alkanoyl, carbamoyl, alkoxy, alkylthio,
alkylamino, (alkyl).sub.2amino, alkylsulfinyl, alkylsulfonyl,
arylsulfonyl, carbocyclyl, aryl, or heterocyclyl, wherein R.sup.15
is optionally substituted with one or more, the same or different,
R.sup.16; and
[0289] R.sup.16 is halogen, nitro, cyano, hydroxy,
trifluoromethoxy, trifluoromethyl, amino, formyl, carboxy,
carbamoyl, mercapto, sulfamoyl, methyl, ethyl, methoxy, ethoxy,
acetyl, acetoxy, methylamino, ethylamino, dimethylamino,
diethylamino, N-methyl-N-ethylamino, acetylamino,
N-methylcarbamoyl, N-ethylcarbamoyl, N,N-dimethylcarbamoyl,
N,N-diethylcarbamoyl, N-methyl-N-ethylcarbamoyl, methylthio,
ethylthio, methylsulfinyl, ethylsulfinyl, mesyl, ethylsulfonyl,
methoxycarbonyl, ethoxycarbonyl, N-methylsulfamoyl,
N-ethylsulfamoyl, N,N-dimethylsulfamoyl, N,N-diethylsulfamoyl,
N-methyl-N-ethylsulfamoyl, carbocyclyl, aryl, or heterocyclyl.
[0290] In certain embodiments, R.sup.1 and R.sup.10 are
hydroxy.
[0291] In certain embodiments, R.sup.11 is alkoxy.
[0292] In certain embodiments, R.sup.4 is hydroxy.
[0293] In certain embodiments, R.sup.5 is alkanoyl.
[0294] In certain embodiments, R.sup.8 is (oxan-2-yl)oxy or
(4-amino-5-hydroxy-6-methyl-oxan-2-yl)oxy.
[0295] In certain embodiments, the pazopanib derivatives are
compounds of formula XVI,
##STR00021##
or salts thereof wherein,
[0296] R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.5, R.sup.6,
R.sup.7, R.sup.8, R.sup.9, R.sup.10, R.sup.11, R.sup.12, and
R.sup.13 are each the same or different hydrogen, alkyl, alkenyl,
halogen, nitro, cyano, hydroxy, amino, mercapto, formyl, carboxy,
alkanoyl, carbamoyl, alkoxy, alkylthio, alkylamino,
(alkyl).sub.2amino, alkylsulfinyl, alkylsulfonyl, arylsulfonyl,
carbocyclyl, aryl, or heterocyclyl, wherein each R.sup.1, R.sup.2,
R.sup.3, R.sup.4, R.sup.5, R.sup.6, R.sup.7, R.sup.8, R.sup.9,
R.sup.10, R.sup.11, R.sup.12, and R.sup.13 are optionally
substituted with one or more, the same or different, R.sup.14;
[0297] R.sup.14 is alkyl, halogen, nitro, cyano, hydroxy, amino,
mercapto, formyl, carboxy, alkanoyl, carbamoyl, alkoxy, alkylthio,
alkylamino, (alkyl).sub.2amino, alkylsulfinyl, alkylsulfonyl,
arylsulfonyl, carbocyclyl, aryl, or heterocyclyl, wherein R.sup.14
is optionally substituted with one or more, the same or different,
R.sup.15; and
[0298] R.sup.15 is halogen, nitro, cyano, hydroxy,
trifluoromethoxy, trifluoromethyl, amino, formyl, carboxy,
carbamoyl, mercapto, sulfamoyl, methyl, ethyl, methoxy, ethoxy,
acetyl, acetoxy, methylamino, ethylamino, dimethylamino,
diethylamino, N-methyl-N-ethylamino, acetylamino,
N-methylcarbamoyl, N-ethylcarbamoyl, N,N-dimethylcarbamoyl,
N,N-diethylcarbamoyl, N-methyl-N-ethylcarbamoyl, methylthio,
ethylthio, methylsulfinyl, ethylsulfinyl, mesyl, ethylsulfonyl,
methoxycarbonyl, ethoxycarbonyl, N-methylsulfamoyl,
N-ethylsulfamoyl, N,N-dimethylsulfamoyl, N,N-diethylsulfamoyl,
N-methyl-N-ethylsulfamoyl, carbocyclyl, aryl, or heterocyclyl.
[0299] In certain embodiments, the camptothecin derivative is a
compound of the formula XVII,
##STR00022##
[0300] or salts thereof wherein,
[0301] R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.5, R.sup.6,
R.sup.7, R.sup.8, R.sup.9, R.sup.10, R.sup.11, and R.sup.12 are
each the same or different hydrogen, alkyl, alkenyl, halogen,
nitro, cyano, hydroxy, amino, mercapto, formyl, carboxy, alkanoyl,
carbamoyl, alkoxy, alkylthio, alkylamino, (alkyl).sub.2amino,
alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl, aryl, or
heterocyclyl, wherein each R.sup.1, R.sup.2, R.sup.3, R.sup.4,
R.sup.5, R.sup.6, R.sup.7, R.sup.8, R.sup.9, R.sup.10, R.sup.11,
and R.sup.12 are optionally substituted with one or more, the same
or different, R.sup.13;
[0302] R.sup.13 is alkyl, halogen, nitro, cyano, hydroxy, amino,
mercapto, formyl, carboxy, alkanoyl, carbamoyl, alkoxy, alkylthio,
alkylamino, (alkyl).sub.2amino, alkylsulfinyl, alkylsulfonyl,
arylsulfonyl, carbocyclyl, aryl, or heterocyclyl, wherein R.sup.13
is optionally substituted with one or more, the same or different,
R.sup.14; and
[0303] R.sup.14 is halogen, nitro, cyano, hydroxy,
trifluoromethoxy, trifluoromethyl, amino, formyl, carboxy,
carbamoyl, mercapto, sulfamoyl, methyl, ethyl, methoxy, ethoxy,
acetyl, acetoxy, methylamino, ethylamino, dimethylamino,
diethylamino, N-methyl-N-ethylamino, acetylamino,
N-methylcarbamoyl, N-ethylcarbamoyl, N,N-dimethylcarbamoyl,
N,N-diethylcarbamoyl, N-methyl-N-ethylcarbamoyl, methylthio,
ethylthio, methylsulfinyl, ethylsulfinyl, mesyl, ethylsulfonyl,
methoxycarbonyl, ethoxycarbonyl, N-methylsulfamoyl,
N-ethylsulfamoyl, N,N-dimethylsulfamoyl, N,N-diethylsulfamoyl,
N-methyl-N-ethylsulfamoyl, carbocyclyl, aryl, or heterocyclyl.
[0304] In certain embodiments, the sunitinib derivatives have
formula XVII,
##STR00023##
[0305] or salts thereof wherein,
[0306] R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.5, R.sup.6,
R.sup.7, R.sup.8, R.sup.9, R.sup.10, and R.sup.11 are each the same
or different hydrogen, alkyl, alkenyl, halogen, nitro, cyano,
hydroxy, amino, mercapto, formyl, carboxy, alkanoyl, carbamoyl,
alkoxy, alkylthio, alkylamino, (alkyl).sub.2amino, alkylsulfinyl,
alkylsulfonyl, arylsulfonyl, carbocyclyl, aryl, or heterocyclyl,
wherein each R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.5, R.sup.6,
R.sup.7, R.sup.8, R.sup.9, R.sup.10, and R.sup.11 are optionally
substituted with one or more, the same or different, R.sup.12;
[0307] R.sup.12 is alkyl, halogen, nitro, cyano, hydroxy, amino,
mercapto, formyl, carboxy, alkanoyl, carbamoyl, alkoxy, alkylthio,
alkylamino, (alkyl).sub.2amino, alkylsulfinyl, alkylsulfonyl,
arylsulfonyl, carbocyclyl, aryl, or heterocyclyl, wherein R.sup.12
is optionally substituted with one or more, the same or different,
R.sup.13; and
[0308] R.sup.13 is halogen, nitro, cyano, hydroxy,
trifluoromethoxy, trifluoromethyl, amino, formyl, carboxy,
carbamoyl, mercapto, sulfamoyl, methyl, ethyl, methoxy, ethoxy,
acetyl, acetoxy, methylamino, ethylamino, dimethylamino,
diethylamino, N-methyl-N-ethylamino, acetylamino,
N-methylcarbamoyl, N-ethylcarbamoyl, N,N-dimethylcarbamoyl,
N,N-diethylcarbamoyl, N-methyl-N-ethylcarbamoyl, methylthio,
ethylthio, methylsulfinyl, ethylsulfinyl, mesyl, ethylsulfonyl,
methoxycarbonyl, ethoxycarbonyl, N-methylsulfamoyl,
N-ethylsulfamoyl, N,N-dimethylsulfamoyl, N,N-diethylsulfamoyl,
N-methyl-N-ethylsulfamoyl, carbocyclyl, aryl, or heterocyclyl.
Evaluations of Compound Activity
[0309] BMP-2 and BMP-4 have been established to be important
factors in embryonic skeletal development. BMP receptors are
transmembrane receptors classified as type I or type II based on
sequence homology and contain a Ser/Thr protein kinase domain. BMP
ligand binding to type I receptor (BMPR-I) induces the association
of BMPR-I and BMPR-II receptors, allowing the constitutively
phosphorylated BMPR-II to phosphorylate and activate the latent
BMPR-I. After activation of BMPR-I, the receptor regulated
(R)-Smad1/5/8 is phosphorylated. Phosphorylation of R-Smad releases
it from the receptor complex and forms hetero-complex associating
with common Smad (Co-Smad, Smad4). Subsequently, R-Smad/Co-Smad
complex translocates into the nucleus and regulates the
transcription of target genes by functioning in concert with other
proteins as transcription factors. See FIG. 1. BMP activity is
regulated prior to receptor recognition by the presence of several
structurally distinct extracellular BMP antagonists such as Noggin,
follistatin, sclerostatin, chordin, DCR, BMPMER, cerberus, gremlin,
DAN, and others.
[0310] Structures of the BMPs and its receptors were analyzed to
identify the various residues involved in their interaction. Using
the LUDI de novo design method a large number of compounds were
computationally screened against the binding sites of BMP-receptors
to identify lead chemical compounds that mimic BMP-receptor
interactions. Small molecules were identified that bind to
BMP-receptor binding epitopes or facilitate BMP binding to the
receptor or block noggin binding to BMP. Their activity was
evaluated by determining the potentiation of alkaline phosphatase
activity in sub-optimal doses of BMP in the presence or absence of
exogenously added noggin.
[0311] Data summary for selected compounds is provided in the table
below:
TABLE-US-00001 Fold increase in Fold increase in Noggin-
BMP-Potentiated Inhibition ALP Assay ALP activity over over
Compound BMP control BMP alone control Clotrimazole 7 4 Honokiol 5
16 Tacrolimus 7 5 Fluticasone Propionate 6 3 Linezolid 5 3
Telmisartan 3 3 Retinoic acid 11 2 Spironolactone 3 4 Chlorambucil
3 4 Isotretinoin 30 3 Acitretin 33 3
The fold-increase in Noggin-inhibition assay is based on BMP-alone
control. Further 3 to 5-fold increases were observed when BMP and
Noggin control is compared.
[0312] Data for additions compounds is provided below.
TABLE-US-00002 Fold-increase of BMP-induced Compound ALP (over BMP
alone control) Teniposide 7.0 Mitomycin C 12.0 Cytarabine HCl 2.0
Vinorelbine tartrate 4.0 Mitoxantrone 4.0 Plicamycin 7.0
Vincristine 5.0 Valrubicin 3.0 Doxorubicin 4.0 Pazopanib 2.5
Topotecan 4.0 Decitabine 6.0 Sunitinib 4.0 Daunomycin,
monohydrochloride 7.0 9-aminocamptothecin 10.0
1-(1,1-diphenylprop-2-yn-1-yl)-1H- 7.0 imidazole
[0313] In certain embodiments, the disclosure contemplates
compositions comprising any of the the compounds herein, or
derivatives or substituted forms, such as those in the table, and
uses for any of the applications disclosed herein.
Growth Factors
[0314] In some embodiments, the disclosure relates to the combined
use of growth factor(s) and compounds disclosed herein such as a
compound selected from clotrimazole, honokiol, magnolol,
tacrolimus, pimecrolimus, sirolimus, everolimus, temsirolimus,
spironolactone, fluticasone, fluticasone propionate, fluticasone
furoate, linezolid, telmisartan, chlorambucil, retinol,
isotretinoin, acitretin, etretinate, retinoic acid (tretinoin),
teniposide, mitomycin C, cytarabine, decitabine, vinblastine,
vincristine, vindesine, vinorelbine, valrubicin, doxorubicin,
daunorubicin, epirubicin, idarubicin, mitoxantrone, pixantrone,
plicamycin, pazopanib, topotecan, camptothecin, irinotecan,
sunitinib, derivatives, or salt thereof and one or more growth
factors in bone growth applications. Typically, the growth factor
is a bone morphogenetic proteins (BMPs), including but not limited
to, BMP-1, BMP-2, BMP-2A, BMP-2B, BMP-3, BMP-3b, BMP-4, BMP-5,
BMP-6, BMP-7 (OP-1), BMP-8, BMP-8b, BMP-9, BMP-10, BMP-11, BMP-12,
BMP-13, BMP-14, BMP-15. BMPs act through specific transmembrane
receptors located on cell surface of the target cells.
[0315] Non-limiting examples of additional suitable growth factors
include osteogenin, insulin-like growth factor (IGF)-1, IGF-II,
TGF-beta1, TGF-beta2, TGF-beta3, TGF-beta4, TGF-beta5,
osteoinductive factor (OIF), basic fibroblast growth factor (bFGF),
acidic fibroblast growth factor (aFGF), platelet-derived growth
factor (PDGF), vascular endothelial growth factor (VEGF), growth
hormone (GH), growth and differentiation factors (GDF)-5 through 9,
and osteogenic protein-1 (OP-1). The growth factors may be isolated
from synthetic methods, recombinant sources or may be purified from
a biological sample. Preferably the growth factors are obtained
from a recombinant technology and for clarity certain embodiments
include rhBMP-2, rhBMP-4, rhBMP-6, rhBMP-7, and rhGDF-5, as
disclosed, for example, in the U.S. Pat. Nos. 4,877,864; 5,013,649;
5,661,007; 5,688,678; 6,177,406; 6,432,919; 6,534,268, and
6,858,431, and in Wozney, J. M., et al. (1988) Science,
242(4885):1528-1534 hereby incorporated by reference.
[0316] In a typical embodiment, a graft composition comprises a
matrix, BMP-2, and one or more compound(s) disclosed herein such a
compound selected from clotrimazole, honokiol, magnolol,
tacrolimus, pimecrolimus, sirolimus, everolimus, temsirolimus,
spironolactone, fluticasone, fluticasone propionate, fluticasone
furoate, linezolid, telmisartan, chlorambucil, retinol,
isotretinoin, acitretin, etretinate, retinoic acid (tretinoin),
teniposide, mitomycin C, cytarabine, decitabine, vinblastine,
vincristine, vindesine, vinorelbine, valrubicin, doxorubicin,
daunorubicin, epirubicin, idarubicin, mitoxantrone, pixantrone,
plicamycin, pazopanib, topotecan, camptothecin, irinotecan,
sunitinib, derivatives, or salt thereof in combinations with other
growth factors such as GDF-5. In one embodiment, the matrix
contains an effective amount of a BMP-2 protein, an rhBMP-2
protein, functional fragments thereof, or combinations thereof. For
certain embodiments, the range of concentrations of BMP-2 may be
about 1.0 to 4.0 mg/ml and GDF-5 concentrations may be 0.25 to 4.0
mg/ml. Although a graft matrix may be loaded during manufacturing,
it is typically loaded just prior to implantation.
[0317] The polypeptide of human BMP-2 is 396 amino acids in length,
and its gene is localized to chromosome 20p12. BMP-2 belongs to the
transforming growth factor-beta (TGF-beta) superfamily. The human
amino acid sequence BMP-2 is SEQ ID NO:1 shown below. Amino acids
38-268 are the TGF-beta propeptide domain, and 291-396 are the
TGF-beta family N-terminal domain. Amino acids 283-396 are the
mature peptide. The mature form of BMP-2 contains four potential
N-linked glycosylation sites per polypeptide chain, and four
potential disulfide bridges. (SEQ ID NO: 1) 1 MVAGTRCLLA LLLPQVLLGG
AAGLVPELGR RKFAAASSGR PSSQPSDEVL SEFELRLLSM 61 FGLKQRPTPS
RDAVVPPYML DLYRRHSGQP GSPAPDHRLE RAASRANTVR SFHHEESLEE 121
LPETSGKTTR RFFFNLSSIP TEEFITSAEL QVFREQMQDA LGNNSSFHHR INIYEIIKPA
181 TANSKFPVTR LLDTRLVNQN ASRWESFDVT PAVMRWTAQG HANHGFVVEV
AHLEEKQGVS 241 KRHVRISRSL HQDEHSWSQI RPLLVTFGHD GKGHPLHKRE
KRQAKHKQRK RLKSSCKRHP 301 LYVDFSDVGW NDWIVAPPGY HAFYCHGECP
FPLADHLNST NHAIVQTLVN SVNSKIPKAC 361 CVPTELSAIS MLYLDENEKV
VLKNYQDMVV EGCGCR.
[0318] In one embodiment, bone morphogenetic protein includes one
of the following synthetic peptide fragments of BMP-2: SEQ ID NO: 2
(KIPKASSVPTELSAISTLYLDDD), SEQ ID NO: 3
(CCCCDDDSKIPKASSVPTELSAISTLYL) SEQ ID NO: 4
(C.sub.16H.sub.31O--NH-CCCCGGGSKIPKASSVPTELSAISTLYL) which may be
synthesized by FMOC/tBu solid-phase peptide synthesis.
[0319] BMP-7 also belongs to the TGF-beta superfamily. It induces
cartilage and bone formation. The amino acid sequence of BMP-7 is
SEQ ID NO: 5. (SEQ ID NO: 5) 1 MHVRSLRAAA PHSFVALWAP LFLLRSALAD
FSLDNEVHSS FIHRRLRSQE RREMQREILS 61 ILGLPHRPRP HLQGKHNSAP
MFMLDLYNAM AVEEGGGPGG QGFSYPYKAV FSTQGPPLAS 121 LQDSHFLTDA
DMVMSFVNLV EHDKEFFHPR YHHREFRFDL SKIPEGEAVT AAEFRIYKDY 181
IRERFDNETF RISVYQVLQE HLGRESDLFL LDSRTLWASE EGWLVFDITA TSNHWVVNPR
241 HNLGLQLSVE TLDGQSINPK LAGLIGRHGP QNKQPFMVAF FKATEVHFRS
IRSTGSKQRS 301 QNRSKTPKNQ EALRMANVAE NSSSDQRQAC KKHELYVSFR
DLGWQDWIIA PEGYAAYYCE 361 GECAFPLNSY MNATNHAIVQ TLVHFINPET
VPKPCCAPTQ LNAISVLYFD DSSNVILKKY 421 RNNVVRACGC H. Amino acids 1-29
are a potential signal sequence; 30-431 are the prepropeptide, and
293-431 are the mature protein. The mature form of BMP-7 contains
four potential N-linked glycosylation sites per polypeptide chain,
and four potential disulfide bridges.
Graft Compositions
[0320] In some embodiments, the disclosure relates to a graft
composition comprising growth factor(s) and clotrimazole, honokiol,
magnolol, tacrolimus, pimecrolimus, sirolimus, everolimus,
temsirolimus, spironolactone, fluticasone, fluticasone propionate,
fluticasone furoate, linezolid, telmisartan, chlorambucil, retinol,
isotretinoin, acitretin, etretinate, retinoic acid (tretinoin),
teniposide, mitomycin C, cytarabine, decitabine, vinblastine,
vincristine, vindesine, vinorelbine, valrubicin, doxorubicin,
daunorubicin, epirubicin, idarubicin, mitoxantrone, pixantrone,
plicamycin, pazopanib, topotecan, camptothecin, irinotecan,
sunitinib, derivatives, or salts thereof. In some embodiments,
these compositions can be created from polymers, bone granules, and
ceramics such as calcium phosphates (e.g. hydroxyapatite and
tricalcium phosphate), bioglass, and calcium sulphate.
[0321] Bioglass refers to materials of SiO.sub.2, Na.sub.2O, CaO,
and P.sub.2O, in specific proportions. The proportions differ from
the traditional soda-lime glasses in lower amounts of silica
(typically less than 60 mol %), higher amounts of sodium and
calcium, and higher calcium/phosphorus ratio. A high ratio of
calcium to phosphorus promotes formation of apatite crystals;
calcium and silica ions can act as crystallization nuclei. Some
formulations bind to soft tissues and bone, some only to bone, some
do not form a bond at all and after implantation get encapsulated
with non-adhering fibrous tissue, and others are completely
absorbed overtime. Mixtures of 35-60 mol % SiO.sub.2, 10-50 mol %
CaO, and 5-40 mol % Na.sub.2O bond to bone and some formulations
bond to soft tissues. Mixtures of >50 mol % SiO.sub.2, <10
mol % CaO, <35 mol % Na.sub.2O typically intigrate within a
month. Some CaO may be replaced with MgO and some Na.sub.2O may be
replaced with K.sub.2O. Some CaO can be replaced with
CaF.sub.2.
[0322] In some embodiments, the disclosure relates to a graft
composition comprising growth factor(s) and compounds disclosed
herein such as clotrimazole, honokiol, magnolol, tacrolimus,
pimecrolimus, sirolimus, everolimus, temsirolimus, spironolactone,
fluticasone, fluticasone propionate, fluticasone furoate,
linezolid, telmisartan, chlorambucil, retinol, isotretinoin,
acitretin, etretinate, retinoic acid (tretinoin), teniposide,
mitomycin C, cytarabine, decitabine, vinblastine, vincristine,
vindesine, vinorelbine, valrubicin, doxorubicin, daunorubicin,
epirubicin, idarubicin, mitoxantrone, pixantrone, plicamycin,
pazopanib, topotecan, camptothecin, irinotecan, sunitinib,
derivatives, or salts thereof and/or polysaccharides such as
hyaluronate, cellulose or cellulose derivatives such as, but not
limited to, hydroxypropyl cellulose, methyl cellulose, ethyl
cellulose, and carboxymethyl cellulose. Typically, cellulose
derivates are used in graft compositions that produce a paste or
putty.
[0323] In some embodiments, the disclosure relates to a bone graft
composition comprising a bone morphogenetic protein and
clotrimazole, honokiol, magnolol, tacrolimus, pimecrolimus,
sirolimus, everolimus, temsirolimus, spironolactone, fluticasone,
fluticasone propionate, fluticasone furoate, linezolid,
telmisartan, chlorambucil, retinol, isotretinoin, acitretin,
etretinate, retinoic acid (tretinoin), teniposide, mitomycin C,
cytarabine, decitabine, vinblastine, vincristine, vindesine,
vinorelbine, valrubicin, doxorubicin, daunorubicin, epirubicin,
idarubicin, mitoxantrone, pixantrone, plicamycin, pazopanib,
topotecan, camptothecin, irinotecan, sunitinib, derivatives, or
salt thereof and a graft matrix. The matrix is typically a polymer
designed to hold bone compatible salts, such as calcium phosphates,
for replacement during bone growth. An example is a bovine Type I
collagen embedded with biphasic calcium phosphate granules.
Optionally, matrix compositions may also include one or more agents
that support the formation, development and growth of new bone,
and/or the remodeling thereof. Typical examples of compounds that
function in, such a supportive manner include extracellular
matrix-associated bone proteins such as alkaline phosphatase,
osteocalcin, bone sialoprotein (BSP) and osteocalcin,
phosphoprotein (SPP)-1, type I collagen, fibronectin, osteonectin,
thrombospondin, matrix-gla-protein, SPARC, and osteopontin.
[0324] In certain embodiments, the graft matrix can be made up of a
hydrogel polymer. Typically, a hydrogel is made-up of acrylate
polymers and copolymers substituted with an abundance of
hydrophilic groups, such as terminal hydroxy or carboxyl groups. In
certain embodiments, the graft composition is biodegradable. In
certain embodiments, the matrix comprises homopolymers and
copolymers consisting of gylcolide and lactide. For certain
embodiments, the graft composition comprises a matrix of
hydroxyethylmethacrylate or hydroxymethylmethyacrylate polymers
containing hydroxyapatite in a mineral content approximately that
of human bone. Such a composition may also be made with
crosslinkers comprising an ester, anhydride, orthoester, amide, or
peptide bond. In some embodiments, crosslinkers contain the
following polymers: polyethylene glycol (PEG), polylactic acid,
polyglycolide or combinations thereof.
[0325] In certain embodiments, the graft can be any material that
is porous including those that have macroporosity (mean pore
diameter greater than or equal to 100 .mu.m), mesoporosity (mean
pore diameter less than 100 .mu.m but greater than or equal to 10
.mu.m) and microporosity (mean pore diameter less than 10 .mu.m).
The pores may be of any size, shape or distribution, or within a
predetermined tolerance. In addition, the pores can be
interconnecting or non-interconnecting. In certain embodiments, the
graft composition can be a plurality of porous or non-porous
granules. The specific surface area of the graft can vary. For
example, when the graft is a porous granule, the specific surface
area can range from about 0.1 m.sup.2/g to about 100 m.sup.2/g.
[0326] Suitable polymers useful for preparing the graft include,
but are not limited to, homopolymers or copolymers of monomers
selected from L-lactide; L-lactic acid; D-lactide; D-lactic acid;
glycolide; alpha-hydroxybutyric acid; alpha-hydroxyvaleric acid;
alpha-hydroxyacetic acid; alpha-hydroxycaproic acid;
alpha-hydroxyheptanoic acid; alpha-hydroxydecanoic acid;
alpha-hydroxymyristic acid; alpha-hydroxyoctanoic acid;
alpha-hydroxystearic acid; hydroxybutyrate; hydroxyvalerate;
beta-propiolactide; beta-propiolactic acid; gamma-caprolactone;
beta-caprolactone; epsilon-caprolactone; gamma-butyrolactone;
pivalolactone; tetramethylglycolide; tetramethylglycolic acid;
dimethylglycolic acid; trimethylene carbonate; dioxanone; those
monomers that form liquid crystal polymers; those monomers that
form cellulose; those monomers that form cellulose acetate; those
monomers that form carboxymethylcellulose; those monomers that form
hydroxypropylmethylcellulose; polyurethane precursors including
macrodiols selected from polycaprolactone, poly(ethylene oxide),
poly(ethylene glycol), poly(ethylene adipate), poly(butylene
oxide), and a mixture thereof, isocyanate-functional compounds
selected from hexamethylene diisocyanate, isophorone diisocyanate,
cyclohexane diisocyanate, hydrogenated methylene diphenylene
diisocyanate, and a mixture thereof, and chain extenders selected
from ethylenediamine, 1,4-butanediol, 1,2-butanediol,
2-amino-1-butanol, thiodiethylene diol, 2-mercaptoethyl ether,
3-hexyne-2,5-diol, citric acid, and a mixture thereof, and any
combination of two or more of the foregoing.
[0327] In certain embodiments, the graft composition may contain
one or more antibiotics and/or anti-inflammatory agents. Suitable
antibiotics include, without limitation, nitroimidazole
antibiotics, tetracyclines, penicillins, cephalosporins,
carbopenems, aminoglycosides, macrolide antibiotics, lincosamide
antibiotics, 4-quinolones, rifamycins and nitrofurantoin. Suitable
specific compounds include, without limitation, ampicillin,
amoxicillin, benzylpenicillin, phenoxymethylpenicillin,
bacampicillin, pivampicillin, carbenicillin, cloxacillin,
cyclacillin, dicloxacillin, methicillin, oxacillin, piperacillin,
ticarcillin, flucloxacillin, cefuroxime, cefetamet, cefetrame,
cefixine, cefoxitin, ceftazidime, ceftizoxime, latamoxef,
cefoperazone, ceftriaxone, cefsulodin, cefotaxime, cephalexin,
cefaclor, cefadroxil, cefalothin, cefazolin, cefpodoxime,
ceftibuten, aztreonam, tigemonam, erythromycin, dirithromycin,
roxithromycin, azithromycin, clarithromycin, clindamycin,
paldimycin, lincomycirl, vancomycin, spectinomycin, tobramycin,
paromomycin, metronidazole, tinidazole, ornidazole, amifloxacin,
cinoxacin, ciprofloxacin, difloxacin, enoxacin, fleroxacin,
norfloxacin, ofloxacin, temafloxacin, doxycycline, minocycline,
tetracycline, chlortetracycline, oxytetracycline, methacycline,
rolitetracyclin, nitrofurantoin, nalidixic acid, gentamicin,
rifampicin, amikacin, netilmicin, imipenem, cilastatin,
chloramphenicol, furazolidone, nifuroxazide, sulfadiazin,
sulfametoxazol, bismuth subsalicylate, colloidal bismuth
subcitrate, gramicidin, mecillinam, cloxiquine, chlorhexidine,
dichlorobenzylalcohol, methyl-2-pentylphenol or any combination
thereof.
[0328] Suitable anti-inflammatory compounds include both steroidal
and non-steroidal structures. Suitable non-limiting examples of
steroidal anti-inflammatory compounds are corticosteroids such as
hydrocortisone, cortisol, hydroxyltriamcinolone, alpha-methyl
dexamethasone, dexamethasone-phosphate, beclomethasone
dipropionates, clobetasol valerate, desonide, desoxymethasone,
desoxycorticosterone acetate, dexamethasone, dichlorisone,
diflorasone diacetate, diflucortolone valerate, fluadrenolone,
fluclorolone acetonide, fludrocortisone, flumethasone pivalate,
fluosinolone acetonide, fluocinonide, flucortine butylesters,
fluocortolone, fluprednidene (fluprednylidene) acetate,
flurandrenolone, halcinonide, hydrocortisone acetate,
hydrocortisone butyrate, methylprednisolone, triamcinolone
acetonide, cortisone, cortodoxone, flucetonide, fludrocortisone,
difluorosone diacetate, fluradrenolone, fludrocortisone,
diflurosone diacetate, fluocinolone, fluradrenolone acetonide,
medrysone, amcinafel, amcinafide, betamethasone and the balance of
its esters, chloroprednisone, chlorprednisone acetate,
clocortelone, clescinolone, dichlorisone, diflurprednate,
flucloronide, flunisolide, fluoromethalone, fluperolone,
fluprednisolone, hydrocortisone valerate, hydrocortisone
cyclopentylpropionate, hydrocortamate, meprednisone, paramethasone,
prednisolone, prednisone, beclomethasone dipropionate,
triamcinolone. Mixtures of the above steroidal anti-inflammatory
compounds may also be used.
[0329] Non-limiting examples of non-steroidal anti-inflammatory
compounds include nabumetone, celecoxib, etodolac, nimesulide,
apasone, gold, oxicams, such as piroxicam, isoxicam, meloxicam,
tenoxicam, sudoxicam, the salicylates, such as aspirin, disalcid,
benorylate, trilisate, safapryn, solprin, diflunisal, and fendosal;
the acetic acid derivatives, such as diclofenac, fenclofenac,
indomethacin, sulindac, tolmetin, isoxepac, furofenac, tiopinac,
zidometacin, acematacin, fentiazac, zomepirac, clindanac, oxepinac,
felbinac, and ketorolac; the fenamates, such as mefenamic,
meclofenamic, flufenamic, niflumic, and tolfenamic acids; the
propionic acid derivatives, such as ibuprofen, naproxen,
benoxaprofen, flurbiprofen, ketoprofen, fenoprofen, fenbufen,
indopropfen, pirprofen, carprofen, oxaprozin, pranoprofen,
miroprofen, tioxaprofen, suprofen, alminoprofen, and tiaprofenic;
and the pyrazoles, such as phenylbutazone, oxyphenbutazone,
feprazone, azapropazone, and trimethazone.
[0330] In certain embodiments, compounds disclosed herein can be
combined with the graft preoperatively as well as
intra-operatively. Where the compound is combined preoperatively,
it can be combined with the graft as part of a manufacturing
process where the compound could be applied to the scaffold in a
buffered solution and then subsequently lyophilized or air dried.
The compound may also be applied by spray drying or other coating
methods. The graft could then be subsequently packaged and
sterilized. Where the compound is combined intra-operatively with
the graft, the graft can be dipped or coated with a buffered
solution including the compound and then applied to the bone site
to be repaired.
[0331] In certain embodiments, the graft can further include an
osteogenic material to provide a viable cell population to the bone
repair site. The osteogenic material can be obtained from both
autogenic sources as well as allogenic sources, such as cadaveric
sources or tissue banks Suitable osteogenic material can include,
for example, viable cell sources such as stem cells, multipotent
cells, pluripotent cells, osteoprogenitor cells, pre-osteoblasts,
mature osteoblasts, and blends and mixtures thereof.
[0332] In certain embodiments, the osteogenic material is obtained
from autogenic and/or allogenic human bone marrow, and according to
another embodiment, the osteogenic material is obtained from
autogenic and/or allogenic human lipoaspirate. Both the bone marrow
and lipoaspirate can be processed to further enhance the desired
cell population for example by filtration, separation and/or
concentration. In order to preserve the viability of the cell
population of the osteogenic material, the osteogenic material is
typically combined with the graft matrix and osteoinductive
material at or near the time of the implantation procedure.
Methods
[0333] Bone grafting is possible because bone tissue, unlike most
other tissues, has the ability to regenerate if provided the space
into which to grow with appropriate chemical signals. With regard
to synthetic grafts, as native bone grows, it typically replaces
most or all of the artificial graft material, resulting in an
integrated region of new bone. However, with regard to certain
embodiments of the disclosure, it is not intended that new bone
must remove all artificial material. In addition, with regard to
certain embodiments of the disclosure, it is not intended that
graft location need contact any other bone of the skeletal
system.
[0334] In certain embodiments, the disclosure relates to a method
of forming bone comprising implanting a graft composition
comprising a compound disclosed herein such as clotrimazole,
honokiol, magnolol, tacrolimus, pimecrolimus, sirolimus,
everolimus, temsirolimus, spironolactone, fluticasone, fluticasone
propionate, fluticasone furoate, linezolid, telmisartan,
chlorambucil, retinol, isotretinoin, acitretin, etretinate,
retinoic acid (tretinoin), teniposide, mitomycin C, cytarabine,
decitabine, vinblastine, vincristine, vindesine, vinorelbine,
valrubicin, doxorubicin, daunorubicin, epirubicin, idarubicin,
mitoxantrone, pixantrone, plicamycin, pazopanib, topotecan,
camptothecin, irinotecan, sunitinib, derivatives, or salts thereof,
in a subject. In certain embodiments, the disclosure relates to
methods of forming bone comprising implanting a graft composition
comprising a bone morphogenetic protein and compound(s) disclosed
herein, such as clotrimazole, honokiol, magnolol, tacrolimus,
pimecrolimus, sirolimus, everolimus, temsirolimus, spironolactone,
fluticasone, fluticasone propionate, fluticasone furoate,
linezolid, telmisartan, chlorambucil, retinol, isotretinoin,
acitretin, etretinate, retinoic acid (tretinoin), teniposide,
mitomycin C, cytarabine, decitabine, vinblastine, vincristine,
vindesine, vinorelbine, valrubicin, doxorubicin, daunorubicin,
epirubicin, idarubicin, mitoxantrone, pixantrone, plicamycin,
pazopanib, topotecan, camptothecin, irinotecan, sunitinib,
derivatives, or salts thereof, in a subject. The graft may be the
result of a void created by surgical removal or created as a result
of an attempt to correct a physical abnormality of a bone, such as
but not limited to, cranial bones; frontal, parietal, temporal,
occipital, sphenoid, ethmoid; facial bones; mandible, maxilla,
palatine, zygomatic, nasal, lacrimal, vomer, inferior nasal
conchae; shoulder girdle; scapula or shoulder blade, clavicle or
collarbone; in the thorax; sternum, manubrium, gladiolus, and
xiphoid process, ribs; in the vertebral column; cervical vertebrae,
thoracic vertebrae; lumbar vertebrae; in the arms, humerus, radius,
ulna; in the pelvis; coccyx; sacrum, hip bone (innominate bone or
coxal bone); in the legs; femur, patella, tibia, and fibula. It is
contemplated that the graft may be added for cosmetic purposes,
e.g., cheek augmentation. In the case of a broken bone or removal
of a bone during surgery, it may be desirable to secure movement of
bone structure with a fixation system and remove the system after
bone forms in the implanted graft.
[0335] With regard to prostheses, it may be desirable to grow bone
between existing bone and an implanted device, or in preparation of
an implanted device, such as in the case of a hip replacement, knee
replacement, and dental implant, i.e., artificial tooth root used
to support restorations that resemble a tooth or group of
teeth.
[0336] In some embodiments, the disclosure relates to
three-dimensional structures made of biocompatible and
biodegradable bone graft materials in the shape of the bone infused
with compositions disclosed herein to promote bone growth. Implants
can be used to support a number of prostheses. A typical implant
consists of a titanium device. In certain embodiments, the graft
compositions disclosed herein contain implants.
[0337] With regard to a sinus augmentation or alveolar ridge
augmentation, surgery may be performed as an outpatient under
general anesthesia, oral conscious sedation, nitrous oxide
sedation, intravenous sedation or under local anesthesia. Bone
grafting is used in cases where there is a lack of adequate
maxillary or mandibular bone in terms of depth or thickness.
Sufficient bone is needed in three dimensions to securely integrate
with the root-like implant. Improved bone height is important to
assure ample anchorage of the root-like shape of the implant.
[0338] In a typical procedure, the clinician creates a large flap
of the gingiva or gum to fully expose the bone at the graft site,
performs one or several types of block and onlay grafts in and on
existing bone, then installs a membrane designed to repel unwanted
infection-causing bacteria. Then the mucosa is carefully sutured
over the site. Together with a course of systemic antibiotics and
topical antibacterial mouth rinses, the graft site is allowed to
heal. The bone graft produces live vascular bone and is therefore
suitable as a foundation for the dental implants.
[0339] In certain embodiments, the disclosure relates to methods of
performing spinal fusion using compositions disclosed herein.
Typically this procedure is used to eliminate the pain caused by
abnormal motion of the vertebrae by immobilizing the vertebrae
themselves. Spinal fusion is often done in the lumbar region of the
spine, but the term is not intended to be limited to method of
fusing lumbar vertebrae. Patients desiring spinal fusion may have
neurological deficits or severe pain which has not responded to
conservative treatment. Conditions where spinal fusion may be
considered include, but are not limited to, degenerative disc
disease, spinal disc herniation, discogenic pain, spinal tumor,
vertebral fracture, scoliosis, kyphosis (i.e, Scheuermann's
disease), spondylolisthesis, or spondylosis.
[0340] In certain embodiments, different methods of lumbar spinal
fusion may be used in conjunction with each other. In one method,
one places the bone graft between the transverse processes in the
back of the spine. These vertebrae are fixed in place with screws
and/or wire through the pedicles of each vertebra attaching to a
metal rod on each side of the vertebrae. In another method, one
places the bone graft between the vertebra in the area usually
occupied by the intervertebral disc. In preparation for the spinal
fusion, the disc is removed entirely. A device may be placed
between the vertebrae to maintain spine alignment and disc height.
The intervertebral device may be made from either plastic or
titanium or other suitable material. The fusion then occurs between
the endplates of the vertebrae. Using both types of fusion are
contemplated.
[0341] In certain embodiments, the disclosure relates to methods of
growing bone at a desired area by locally administering a
pharmaceutical formulation containing a compound disclosed herein
such as clotrimazole, honokiol, magnolol, tacrolimus, pimecrolimus,
sirolimus, everolimus, temsirolimus, spironolactone, fluticasone,
fluticasone propionate, fluticasone furoate, linezolid,
telmisartan, chlorambucil, retinol, isotretinoin, acitretin,
etretinate, retinoic acid (tretinoin), teniposide, mitomycin C,
cytarabine, decitabine, vinblastine, vincristine, vindesine,
vinorelbine, valrubicin, doxorubicin, daunorubicin, epirubicin,
idarubicin, mitoxantrone, pixantrone, plicamycin, pazopanib,
topotecan, camptothecin, irinotecan, sunitinib, derivatives, or
salts thereof, to the area of desired bone growth with or without
the presence of a graft composition or bone growth matrix
optionally in combination with a growth factor such as a bone
morphogenetic protein. In certain embodiments, the disclosure
contemplates administering the pharmaceutical compositions between
vertebra, e.g., in the area usually occupied by in the
intervertebral disc, to form a spinal fusion.
Cartilage Repair
[0342] Cartilage is typically composed of chondroblasts, Type I and
Type II collagen fibers, elastin fibers, and proteoglycans. Typical
locations within the human body to find cartilage are the joints
between bones, the ear, the nose, the elbow, the knee, the ankle,
and the intervertebral discs. Cartilage can become damaged because
of trauma or disease. In some embodiments, the disclosure relates
to using compounds disclosed herein such as clotrimazole, honokiol,
magnolol, tacrolimus, pimecrolimus, sirolimus, everolimus,
temsirolimus, spironolactone, fluticasone, fluticasone propionate,
fluticasone furoate, linezolid, telmisartan, chlorambucil, retinol,
isotretinoin, acitretin, etretinate, retinoic acid (tretinoin),
teniposide, mitomycin C, cytarabine, decitabine, vinblastine,
vincristine, vindesine, vinorelbine, valrubicin, doxorubicin,
daunorubicin, epirubicin, idarubicin, mitoxantrone, pixantrone,
plicamycin, pazopanib, topotecan, camptothecin, irinotecan,
sunitinib, derivatives, or salts thereof for the repair or
regeneration of cartilage such as articular cartilage repair or
regeneration or intervertebral disc cartilage repair or
regeneration.
[0343] Articular cartilage repair is typically done to restore the
cartilage on the surface of a bone, i.e., hyaline cartilage.
Osteochondrial autografts or allografts may be performed. In
certain embodiments, the disclosure contemplates methods of
cartilage repair comprising transplanting sections of cartilage
and/or bone to a location where cartilage and/or bone was removed
and placing a compound disclosed herein such as herein such as
clotrimazole, honokiol, magnolol, tacrolimus, pimecrolimus,
sirolimus, everolimus, temsirolimus, spironolactone, fluticasone,
fluticasone propionate, fluticasone furoate, linezolid,
telmisartan, chlorambucil, retinol, isotretinoin, acitretin,
etretinate, retinoic acid (tretinoin), teniposide, mitomycin C,
cytarabine, decitabine, vinblastine, vincristine, vindesine,
vinorelbine, valrubicin, doxorubicin, daunorubicin, epirubicin,
idarubicin, mitoxantrone, pixantrone, plicamycin, pazopanib,
topotecan, camptothecin, irinotecan, sunitinib, derivatives, or
salts thereof about the surrounding area, e.g., by injections at
the site of transplantation. Bone with its cartilage covering may
be removed from the same or a different joint and replanted into
the hole left from removing degraded bone and cartilage. The
transplanted bone and cartilage are typically taken from areas of
low stress.
[0344] In autologous chondrocyte implantation, cartilage cells are
typically extracted arthroscopically from normal articular
cartilage of the subject that is located in a nonload-bearing area,
e.g., the intercondylar notch or the superior ridge of the femoral
condyles, and the cells are replicated, in vitro, in the presence
of growth factors. In certain embodiments, the disclosure relates
to replicating cartilage cells comprising mixing hyaline cartilage
and a compound disclosed herein such as clotrimazole, honokiol,
magnolol, tacrolimus, pimecrolimus, sirolimus, everolimus,
temsirolimus, spironolactone, fluticasone, fluticasone propionate,
fluticasone furoate, linezolid, telmisartan, chlorambucil, retinol,
isotretinoin, acitretin, etretinate, retinoic acid (tretinoin),
teniposide, mitomycin C, cytarabine, decitabine, vinblastine,
vincristine, vindesine, vinorelbine, valrubicin, doxorubicin,
daunorubicin, epirubicin, idarubicin, mitoxantrone, pixantrone,
plicamycin, pazopanib, topotecan, camptothecin, irinotecan,
sunitinib, derivatives, or salts thereof, under conditions such
that the cartilage cells replicate. Typically this is done by
adding other growth factors to the cartilage replicating medium,
e.g., cartilage-derived morphogenetic proteins and/or BMP proteins.
The replicated chondrocytes are implanted to the desired area,
e.g., injected about the site of the area for repair optionally in
combination with either a membrane or a matrix comprising growth
factors such as a CDMP, BMP protein or a compound disclosed
herein.
[0345] Repair of articular cartilage may be performed by marrow
stimulating procedures sometimes referred to as microfracture
surgery. Damaged cartilage is typically ablated by, e.g., drilling
or pounding, exposing the underlying bone--sometimes referred to as
a microfracture. The subchondal bone typically generates a blood
clot followed by cartilage regeneration. In some embodiments the
disclosure relates to methods of generating cartilage by disrupting
bone underlying articular cartilage and placing a compound
disclosed herein about the area of disruption, e.g., by injecting
compounds disclosed herein such as clotrimazole, honokiol,
magnolol, tacrolimus, pimecrolimus, sirolimus, everolimus,
temsirolimus, spironolactone, fluticasone, fluticasone propionate,
fluticasone furoate, linezolid, telmisartan, chlorambucil, retinol,
isotretinoin, acitretin, etretinate, retinoic acid (tretinoin),
teniposide, mitomycin C, cytarabine, decitabine, vinblastine,
vincristine, vindesine, vinorelbine, valrubicin, doxorubicin,
daunorubicin, epirubicin, idarubicin, mitoxantrone, pixantrone,
plicamycin, pazopanib, topotecan, camptothecin, irinotecan,
sunitinib, derivatives, or salts thereof about the site of
disrupted bone for the improved repair or regeneration of cartilage
optionally in combination with a growth factor such as a CDMP
and/or BMP protein. Alternatively it is contemplated that the
compounds are administered to the subject in a pharmaceutical
composition before, during or after the procedure. In another
alternative, it is contemplated that a collagen matrix is implanted
at the site of the exposed underlying bone to improve chondrogenic
differentiation of mesenchymal stem cells. It is also contemplated
that the subject may optionally be postoperative injected with
compounds disclosed herein, hyaluronic acid, and/or mesenchymal
stem cells, e.g., obtained from autologous peripheral blood
progenitor cells.
[0346] Inflammation of the synovial membrane in a joint causes
swelling and joint surface destruction. Removing excess fluid and
material by a lavage or debridement frequently resolves arthritic
knee inflammation and pain. In certain embodiments, the disclosure
relates to the use of compounds disclosed herein such as
clotrimazole, honokiol, magnolol, tacrolimus, pimecrolimus,
sirolimus, everolimus, temsirolimus, spironolactone, fluticasone,
fluticasone propionate, fluticasone furoate, linezolid,
telmisartan, chlorambucil, retinol, isotretinoin, acitretin,
etretinate, retinoic acid (tretinoin), teniposide, mitomycin C,
cytarabine, decitabine, vinblastine, vincristine, vindesine,
vinorelbine, valrubicin, doxorubicin, daunorubicin, epirubicin,
idarubicin, mitoxantrone, pixantrone, plicamycin, pazopanib,
topotecan, camptothecin, irinotecan, sunitinib, derivatives, or
salts thereof before, during, or after a lavage or debridement
inside a joint, e.g., arthroscopic lavage, arthroscopic
debridement. In arthroscopic debridement, joint material or
degenerative cartilage it typically removed by injecting a fluid
and removing it with a vacuum.
[0347] An intervertebral disc (IVD) is found in between two
vertebrae. The IVD contains different tissue types such as the
annulus fibrosus (AF), the nucleus pulposus (NP), and end-plates.
The AF is made up of mainly collagen type I. The amount of collagen
type I decreases and collagen type II increase gradually nearer the
NP which is mostly collagen type II dispersed within a
proteoglycan-rich gelatinous matrix surrounding the NP.
[0348] Porous silk scaffolds may be used for a variety of
tissue-engineering applications, such as the regeneration of bone
and cartilage. Removal of sericin from silk reduces immunogenic
responses. Silk may form a desired sponge-like structure by
freeze-drying a silk solution. Bone marrow mesenchymal stem cells
(BMSC) may be incorporated into porous silk scaffolds wrapped
around a silicone NP substitute to form an artificial
intervertebral disc. In certain embodiments, it is contemplated
that compounds disclosed herein may be used to generate a matrix of
annulus fibrosus by mixing with mesenchymal stem cells and growth
factors. In certain embodiments, the disclosure contemplates
implanting a fabricated intervertebral disc into a subject wherein
the disc comprises annulus fibrosus tissue and placing a compound
disclosed herein about the site of the implant location, e.g., by
injection, optionally in combination with a growth factor such as a
cartilage-derived morphogenetic protein (CDMP), e.g., CDMP-1 or
CDMP-2, and/or bone morphogenetic proteins, e.g., BMP-7 or BMP-14.
The fabricated disc may comprise a NP area with a hydrogel
polymer/copolymer matrix or a collagen and/or hyaluronan and/or
chondroitin-6-sulfate copolymer. A variety of stem cells, such as
mesenchymal stem cells, synovium-derived stem cells (SDSCs), or
notochord cells, may be used for rejuvenation of NP cells.
Therapeutic Applications
[0349] In some embodiments, the disclosure relates to
pharmaceutical compositions comprising compounds disclosed herein
for therapeutic applications. In some embodiments, the disclosure
relates to methods of treating bone degenerative disorders, such as
osteoporosis, osteitis deformans ("Paget's disease of bone"), bone
metastasis (with or without hypercalcaemia), multiple myeloma,
primary hyperparathyroidism, or osteogenesis imperfecta.
Osteoporosis is a disease of bones that leads to an increased risk
of fracture. In osteoporosis, the bone mineral density (BMD) is
reduced, bone microarchitecture is disrupted, and the amount and
variety of proteins in bone is altered. Osteoporosis is most common
in women after menopause, when it is called postmenopausal
osteoporosis, but may also develop in men, and may occur in anyone
in the presence of particular hormonal disorders and other chronic
diseases or as a result of medications, specifically
glucocorticoids, when the disease is called steroid- or
glucocorticoid-induced osteoporosis (SIOP or GIOP).
[0350] Osteoporotic fractures are those that occur in situations
where healthy people would not normally break a bone; they are
therefore regarded as fragility fractures. Typical fragility
fractures occur in the vertebral column, rib, hip and wrist. The
diagnosis of osteoporosis can be made using conventional
radiography by measuring the bone mineral density (BMD).
[0351] In some embodiments, the disclosure relates to treating bone
degenerative disorders by administering pharmaceutical composition
described herein in combination with other agents, such as calcium
carbonate and calcium citrate, vitamine D, cholecalciferol,
1,25-dihydroxycholecalciferol, calcitriol, estrogen, testosterone,
raloxifene, pamidronate, neridronate, olpadronate, alendronate
(Fosamax), ibandronate (Boniva), risedronate (Actonel), zoledronate
(Zometa, Aclasta), etidronate (Didronel), clodronate (Bonefos,
Loron), or tiludronate (Skelid).
[0352] In some embodiments, the disclosure relates to using
compounds disclosed herein such as clotrimazole, honokiol,
magnolol, tacrolimus, pimecrolimus, sirolimus, everolimus,
temsirolimus, spironolactone, fluticasone, fluticasone propionate,
fluticasone furoate, linezolid, telmisartan, chlorambucil, retinol,
isotretinoin, acitretin, etretinate, retinoic acid (tretinoin),
teniposide, mitomycin C, cytarabine, decitabine, vinblastine,
vincristine, vindesine, vinorelbine, valrubicin, doxorubicin,
daunorubicin, epirubicin, idarubicin, mitoxantrone, pixantrone,
plicamycin, pazopanib, topotecan, camptothecin, irinotecan,
sunitinib, derivatives, or salts thereof in the treatment of
chondrodystrophies. Typically an effective amount of a
pharmaceutical composition comprising the compound is administered
to a subject diagnosed with, at risk of, or exhibiting symptoms of
osteoarthritis, achondroplasia, costochondrits, relapsing
polychondritis, or articular cartilage damage. The pharmaceutical
compositions may provide pain relief or slow down the progression
of damage delaying joint replacement (knee replacement)
surgery.
[0353] In some embodiments, the disclosure relates to using
compounds disclosed herein such as clotrimazole, honokiol,
magnolol, tacrolimus, pimecrolimus, sirolimus, everolimus,
temsirolimus, spironolactone, fluticasone, fluticasone propionate,
fluticasone furoate, linezolid, telmisartan, chlorambucil, retinol,
isotretinoin, acitretin, etretinate, retinoic acid (tretinoin),
teniposide, mitomycin C, cytarabine, decitabine, vinblastine,
vincristine, vindesine, vinorelbine, valrubicin, doxorubicin,
daunorubicin, epirubicin, idarubicin, mitoxantrone, pixantrone,
plicamycin, pazopanib, topotecan, camptothecin, irinotecan,
sunitinib, derivatives, or salts thereof in the treatment of a
degenerative intervertebrial disc. Typically an effective amount of
a pharmaceutical composition comprising the compound is
administered to a subject diagnosed with, at risk of, or exhibiting
symptoms of a degenerative disc. The compositions may be
administered orally or injected directly into an intervertebral
disc (IVD), e.g., into the annulus fibrosus (AF) and/or the nucleus
pulposus (NP) optionally in combination with a growth factor such
as a cartilage-derived morphogenetic protein (CDMP), e.g., CDMP-1
or CDMP-2, or a bone morphogenetic protein, e.g., BMP-7 or
BMP-14.
Formulations
[0354] Pharmaceutical compositions disclosed herein may be in the
form of pharmaceutically acceptable salts, as generally described
below. Some preferred, but non-limiting examples of suitable
pharmaceutically acceptable organic and/or inorganic acids are
hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid,
acetic acid and citric acid, as well as other pharmaceutically
acceptable acids known per se (for which reference is made to the
references referred to below).
[0355] When the compounds of the disclosure contain an acidic group
as well as a basic group, the compounds of the disclosure may also
form internal salts, and such compounds are within the scope of the
disclosure. When the compounds of the disclosure contain a
hydrogen-donating heteroatom (e.g. NH), the disclosure also covers
salts and/or isomers formed by transfer of said hydrogen atom to a
basic group or atom within the molecule.
[0356] Pharmaceutically acceptable salts of the compounds include
the acid addition and base salts thereof. Suitable acid addition
salts are formed from acids which form non-toxic salts. Examples
include the acetate, adipate, aspartate, benzoate, besylate,
bicarbonate/carbonate, bisulphate/sulphate, borate, camsylate,
citrate, cyclamate, edisylate, esylate, formate, fumarate,
gluceptate, gluconate, glucuronate, hexafluorophosphate, hibenzate,
hydrochloride/chloride, hydrobromide/bromide, hydroiodide/iodide,
isethionate, lactate, malate, maleate, malonate, mesylate,
methylsulphate, naphthylate, 2-napsylate, nicotinate, nitrate,
orotate, oxalate, palmitate, pamoate, phosphate/hydrogen
phosphate/dihydrogen phosphate, pyroglutamate, saccharate,
stearate, succinate, tannate, tartrate, tosylate, trifluoroacetate
and xinofoate salts. Suitable base salts are formed from bases
which form non-toxic salts. Examples include the aluminium,
arginine, benzathine, calcium, choline, diethylamine, diolamine,
glycine, lysine, magnesium, meglumine, olamine, potassium, sodium,
tromethamine and zinc salts. Hemisalts of acids and bases may also
be formed, for example, hemisulphate and hemicalcium salts. For a
review on suitable salts, see Handbook of Pharmaceutical Salts:
Properties, Selection, and Use by Stahl and Wermuth (Wiley-VCH,
2002), incorporated herein by reference.
[0357] The compounds described herein may be administered in the
form of prodrugs. A prodrug can include a covalently bonded carrier
which releases the active parent drug when administered to a
mammalian subject. Prodrugs can be prepared by modifying functional
groups present in the compounds in such a way that the
modifications are cleaved, either in routine manipulation or in
vivo, to the parent compounds. Prodrugs include, for example,
compounds wherein a hydroxy group is bonded to any group that, when
administered to a mammalian subject, cleaves to form a free hydroxy
group. Examples of prodrugs include, but are not limited to,
acetate, formate, and benzoate derivatives of alcohol functional
groups in the compounds. Methods of structuring a compound as
prodrugs can be found in the book of Testa and Mayer, Hydrolysis in
Drug and Prodrug Metabolism, Wiley (2006). Typical prodrugs form
the active metabolite by transformation of the prodrug by
hydrolytic enzymes, the hydrolysis of amide, lactams, peptides,
carboxylic acid esters, epoxides or the cleavage of esters of
inorganic acids.
[0358] Pharmaceutical compositions for use in the present
disclosure typically comprise an effective amount of a compound and
a suitable pharmaceutical acceptable carrier. The preparations may
be prepared in a manner known per se, which usually involves mixing
the at least one compound according to the disclosure with the one
or more pharmaceutically acceptable carriers, and, if desired, in
combination with other pharmaceutical active compounds, when
necessary under aseptic conditions. Reference is made to U.S. Pat.
No. 6,372,778, U.S. Pat. No. 6,369,086, U.S. Pat. No. 6,369,087 and
U.S. Pat. No. 6,372,733 and the further references mentioned above,
as well as to the standard handbooks, such as the latest edition of
Remington's Pharmaceutical Sciences.
[0359] Generally, for pharmaceutical use, the compounds may be
formulated as a pharmaceutical preparation comprising at least one
compound and at least one pharmaceutically acceptable carrier,
diluent or excipient and/or adjuvant, and optionally one or more
further pharmaceutically active compounds.
[0360] The pharmaceutical preparations of the disclosure are
preferably in a unit dosage form, and may be suitably packaged, for
example in a box, blister, vial, bottle, sachet, ampoule or in any
other suitable single-dose or multi-dose holder or container (which
may be properly labeled); optionally with one or more leaflets
containing product information and/or instructions for use.
Generally, such unit dosages will contain between 1 and 1000 mg,
and usually between 5 and 500 mg, of the at least one compound of
the disclosure, e.g. about 10, 25, 50, 100, 200, 300 or 400 mg per
unit dosage.
[0361] The compounds can be administered by a variety of routes
including the oral, ocular, rectal, transdermal, subcutaneous,
intravenous, intramuscular or intranasal routes, depending mainly
on the specific preparation used. The compound will generally be
administered in an "effective amount", by which is meant any amount
of a compound that, upon suitable administration, is sufficient to
achieve the desired therapeutic or prophylactic effect in the
subject to which it is administered. Usually, depending on the
condition to be prevented or treated and the route of
administration, such an effective amount will usually be between
0.01 to 1000 mg per kilogram body weight of the patient per day,
more often between 0.1 and 500 mg, such as between 1 and 250 mg,
for example about 5, 10, 20, 50, 100, 150, 200 or 250 mg, per
kilogram body weight of the patient per day, which may be
administered as a single daily dose, divided over one or more daily
doses. The amount(s) to be administered, the route of
administration and the further treatment regimen may be determined
by the treating clinician, depending on factors such as the age,
gender and general condition of the patient and the nature and
severity of the disease/symptoms to be treated. Reference is again
made to U.S. Pat. No. 6,372,778, U.S. Pat. No. 6,369,086, U.S. Pat.
No. 6,369,087, and U.S. Pat. No. 6,372,733 and the further
references mentioned above, as well as to the standard handbooks,
such as the latest edition of Remington's Pharmaceutical
Sciences.
[0362] For an oral administration form, the compound can be mixed
with suitable additives, such as excipients, stabilizers or inert
diluents, and brought by means of the customary methods into the
suitable administration forms, such as tablets, coated tablets,
hard capsules, aqueous, alcoholic, or oily solutions. Examples of
suitable inert carriers are gum arabic, magnesia, magnesium
carbonate, potassium phosphate, lactose, glucose, or starch, in
particular, corn starch. In this case, the preparation can be
carried out both as dry and as moist granules. Suitable oily
excipients or solvents are vegetable or animal oils, such as
sunflower oil or cod liver oil. Suitable solvents for aqueous or
alcoholic solutions are water, ethanol, sugar solutions, or
mixtures thereof. Polyethylene glycols and polypropylene glycols
are also useful as further auxiliaries for other administration
forms. As immediate release tablets, these compositions may contain
microcrystalline cellulose, dicalcium phosphate, starch, magnesium
stearate and lactose and/or other excipients, binders, extenders,
disintegrants, diluents and lubricants known in the art.
[0363] When administered by nasal aerosol or inhalation, the
compositions may be prepared according to techniques well-known in
the art of pharmaceutical formulation and may be prepared as
solutions in saline, employing benzyl alcohol or other suitable
preservatives, absorption promoters to enhance bioavailability,
fluorocarbons, and/or other solubilizing or dispersing agents known
in the art. Suitable pharmaceutical formulations for administration
in the form of aerosols or sprays are, for example, solutions,
suspensions or emulsions of the compounds of the disclosure or
their physiologically tolerable salts in a pharmaceutically
acceptable solvent, such as ethanol or water, or a mixture of such
solvents. If required, the formulation can also additionally
contain other pharmaceutical auxiliaries such as surfactants,
emulsifiers and stabilizers as well as a propellant.
[0364] For subcutaneous or intravenous administration, the
compounds, if desired with the substances customary therefore such
as solubilizers, emulsifiers or further auxiliaries are brought
into solution, suspension, or emulsion. The compounds of formula I
can also be lyophilized and the lyophilizates obtained used, for
example, for the production of injection or infusion preparations.
Suitable solvents are, for example, water, physiological saline
solution or alcohols, e.g. ethanol, propanol, glycerol, sugar
solutions such as glucose or mannitol solutions, or mixtures of the
various solvents mentioned. The injectable solutions or suspensions
may be formulated according to known art, using suitable non-toxic,
parenterally-acceptable diluents or solvents, such as mannitol,
1,3-butanediol, water, Ringer's solution or isotonic sodium
chloride solution, or suitable dispersing or wetting and suspending
agents, such as sterile, bland, fixed oils, including synthetic
mono- or diglycerides, and fatty acids, including oleic acid.
[0365] When rectally administered in the form of suppositories, the
formulations may be prepared by mixing the compounds of formula I
with a suitable non-irritating excipient, such as cocoa butter,
synthetic glyceride esters or polyethylene glycols, which are solid
at ordinary temperatures, but liquefy and/or dissolve in the rectal
cavity to release the drug.
[0366] In certain embodiments, it is contemplated that these
compositions can be extended release formulations. Typical extended
release formations utilize an enteric coating. Typically, a barrier
is applied to oral medication that controls the location in the
digestive system where it is absorbed. Enteric coatings prevent
release of medication before it reaches the small intestine.
Enteric coatings may contain polymers of polysaccharides, such as
maltodextrin, xanthan, scleroglucan dextran, starch, alginates,
pullulan, hyaloronic acid, chitin, chitosan and the like; other
natural polymers, such as proteins (albumin, gelatin etc.),
poly-L-lysine; sodium poly(acrylic acid);
poly(hydroxyalkylmethacrylates) (for example poly(hydroxyethyl
methacrylate)); carboxypolymethylene (for example Carbopol.TM.);
carbomer; polyvinyl pyrrolidone; gums, such as guar gum, gum
arabic, gum karaya, gum ghatti, locust bean gum, tamarind gum,
gellan gum, gum tragacanth, agar, pectin, gluten and the like;
poly(vinyl alcohol); ethylene vinyl alcohol; polyethylene glycol
(PEG); and cellulose ethers, such as hydroxymethylcellulose (HMC),
hydroxyethylcellulose (HEC), hydroxypropylcellulose (HPC),
methylcellulose (MC), ethylcellulose (EC), carboxyethylcellulose
(CEC), ethylhydroxyethyl cellulose (EHEC),
carboxymethylhydroxyethylcellulose (CMHEC),
hydroxypropylmethyl-cellulose (HPMC), hydroxypropylethylcellulose
(HPEC) and sodium carboxymethylcellulose (Na CMC); as well as
copolymers and/or (simple) mixtures of any of the above polymers.
Certain of the above-mentioned polymers may further be crosslinked
by way of standard techniques.
[0367] The choice of polymer will be determined by the nature of
the active ingredient/drug that is employed in the composition of
the disclosure as well as the desired rate of release. In
particular, it will be appreciated by the skilled person, for
example in the case of HPMC, that a higher molecular weight will,
in general, provide a slower rate of release of drug from the
composition. Furthermore, in the case of HPMC, different degrees of
substitution of methoxyl groups and hydroxypropoxyl groups will
give rise to changes in the rate of release of drug from the
composition. In this respect, and as stated above, it may be
desirable to provide compositions of the disclosure in the form of
coatings in which the polymer carrier is provided by way of a blend
of two or more polymers of, for example, different molecular
weights in order to produce a particular required or desired
release profile.
[0368] Microspheres of polylactide, polyglycolide, and their
copolymers poly(lactide-co-glycolide) may be used to form
sustained-release protein or compound delivery systems. Proteins
and/or compounds can be entrapped in the poly(lactide-co-glycolide)
microsphere depot by a number of methods, including formation of a
water-in-oil emulsion with water-borne protein and organic
solvent-borne polymer (emulsion method), formation of a
solid-in-oil suspension with solid protein dispersed in a
solvent-based polymer solution (suspension method), or by
dissolving the protein in a solvent-based polymer solution
(dissolution method). One can attach poly(ethylene glycol) to
proteins (PEGylation) to increase the in vivo half-life of
circulating therapeutic proteins and decrease the chance of an
immune response.
Experimental
Compound Screenings
[0369] Compounds were selected by virtual screening for development
based on computational modeling, docking, and in silico screening.
The structural data revealed that the surfaces of BMPs have binding
epitopes for BMPR-IA interaction involving the main chain amide
groups of amino acids L51 and D53 with minor contribution from the
hydrophobic interactions. Two hydrogen bonds are formed between L51
(main chain amide and carbonyl) of BMP-2 and Q86 of BMPR-IA. The
core structure of BMPR-II shares the same pattern of disulfide
connectivity as ActR-II, with disulfide bonds. In an earlier
analysis a hypothetical complex between BMPR-II and BMP2, created
by superimposing BMPR-II in the position of ActR-II in the ternary
complex between ActR-II, BMP2, and BMPR-IA, suggested that BMPR-II
makes similar overall ligand binding contacts to BMP2 as does
ActR-II. From Residues Tyr67, Trp85, and Phe115 of BMPRII are in
the main hydrophobic patch and Lys81, Ser86, Glu93, and Tyr113 are
also important binding determinants. In addition, His87 and Tyr40
confer specificity in BMPR-II ligand binding. Mutational analysis
has identified A34, H39, S88, L90, and L100 residues as binding
determinants of BMP-2 for BMPR-II and ActR-II. See Sebald et al.,
Biol Chem, 2004, 385(8):697-710.
[0370] Crystal structure of BMPR-II was superimposed on to the
ActR-IIB of ternary complex consisting of BMP-2
dimer/BMPR-IAEC/ActR-IIB using a web based tool Superpose. Using
these superposed structures a new hypothetical ternary complex
consisting of BMP-2/BMPR-IAEC/BMPR-II was modelled. The surface
area (A2) for each residue was calculated and the percent solvent
accessible contact area (% SA) of the model ternary complex of
BMP-2/BMPR-IAEC/BMPR-II and individual monomer structures, and the
position (Pos) and name of the amino acid residues in the binding
interface of both the receptors, BMPR-IA and BMPR-II that bind to
BMP-2 were identified.
[0371] LUDI de novo design method of Accelrys Discovery Studios 3.0
and also NCC library of compounds, which contains Phase I, II, and
III drugs, were used against the receptor regions that bind to BMP.
The BMP binding region of receptors is divided into ten sub regions
(with some overlapping residues) for LUDI runs. The binding site
defined as spheres of radius 10 angstrom covering the amino acids
in the regions. The LUDI de novo design method against these ten
regions derived 300 novel small molecules that were chemically
synthesized for cell-based screening assays. LUDI against the
BMPR-IA and BMPR-II regions that bind to BMP-2 were also used to
obtain molecules from NCC library that are drug molecules from
Phase I, II or III.
[0372] The BMP-potentiating activities of compounds may be
evaluated by monitoring several markers of the osteoblastic
phenotype corresponding to various time points during phenotype
differentiation of C2C12 cells towards terminally differentiated
osteoblasts.
Cell Culture
[0373] Mouse C2C12 cells and Dulbecco's modified Eagle's medium
(DMEM) were purchased from ATCC (Manassas, Va.). The non-heat
inactivated fetal bovine serum (FBS) was purchased from HyClone
Laboratories, Inc. (Logan, Utah). The C2C12 cells at passages 5 to
10 were subcultured in T-75 cm.sup.2 flasks in DMEM supplemented
with 10% FBS at 37.degree. C. in 5% CO.sub.2 with humidification.
When the flasks reached 80% confluence, the cells were trypsinized
and seeded in triplicate.
Alkaline Phosphatase (ALP) Assay
[0374] The C2C12 cells were plated at 200,000 cells/well in 6-well
plates and grown overnight in DMEM containing 10% FBS. On day 2,
the culture medium was replaced with DMEM containing 2% FBS and the
cells were treated with various concentrations of compound for 24
hours. On day 3, the culture medium was replaced with DMEM
containing 2% FBS and the cells were treated with 50 ng/ml of BMP-2
and various concentrations of compound for 72 hours. The cells were
washed with phosphate-buffered saline (PBS) and lysed by addition
of lysis buffer (10 mM Tris-HCl pH 8.0, 1 mM MgCl.sub.2 and 1.0%
Triton X-100). The cell lysates were centrifuged for 5 minutes at
13,000.times.g. The supernatant was removed and the aliquots were
assayed for ALP activity and protein amount. The ALP activity was
measured in triplicate using an ALP assay kit (Sigma-Aldrich, St.
Louis, Mo.) in microtiter plates. The protein amount was determined
with Bio-Rad protein assay reagent (Bio-Rad, Hercules, Calif.)
using bovine serum albumin (BSA) as a standard. The ALP activity
(nmoles of p-nitrophenol per ml) was normalized to the protein
amount (nmoles of p-nitrophenol per .mu.g).
BMP-Noggin Competitive Alkaline Phosphatase (ALP) Assay
[0375] The C2C12 cells were plated at 200,000 cells/well in 6-well
plates and grown overnight in DMEM containing 10% FBS. On day 2,
the culture medium was replaced with DMEM containing 2% FBS and the
cells were treated with 90-100 ng/ml of BMP-2 (as noted in each
experiment), various concentrations of compound, and 100-120 ng/ml
of noggin (as noted in each experiment) for 72 hours. The cells
were washed with phosphate-buffered saline (PBS) and lysed by
addition of lysis buffer (10 mM Tris-HCl pH 8.0, 1 mM MgCl.sub.2
and 1.0% Triton X-100). The cell lysates were centrifuged for 5
minutes at 13,000.times.g. The supernatant was removed and the
aliquots were assayed for ALP activity and protein amount. The ALP
activity was measured in triplicate using an ALP assay kit
(Sigma-Aldrich, St. Louis, Mo.) in microtiter plates. The protein
amount was determined with Bio-Rad protein assay reagent (Bio-Rad,
Hercules, Calif.) using bovine serum albumin (BSA) as a standard.
The ALP activity (nmoles of p-nitrophenol per ml) was normalized to
the protein amount (nmoles of p-nitrophenol per .mu.g).
Collagen Disc Implantation with Macrolides in Rat Ectopic Model
[0376] Sprague Dawley rats about 5-6 weeks of age were chest
implanted with a collagen disc (diameter of 1.0 cm and height of
2.0 mm; total volume of 150 cubic millimeters) and doses of
macrolides in the absence of BMP-2. After 4 weeks the rats were
sacrificed and evaluated for bone growth. See table below. De novo
bone formation is demonstrated locally with sirolimus, everolimus,
and tacrolimus. In certain embodiments, the disclosure contemplates
local bone formation for fracture repair, segmental bone defects,
spine fusion, bone grafting, and regional bone enhancement for
osteopenic bones before they fracture (e.g. hip, vertebral body,
etc) by deliver locally to induce local bone formation.
TABLE-US-00003 Animal # Dose with Xray: Drug (mM) Volume Carrier
1..2..3..4 Results Ave. Siro- 0 100 ul collagen 0, 0, 0, 0 No bone
(0 of 4) 0 limus 10 100 ul collagen 0, (1), 0, 0 Bone (1 of 4) 0.25
15 100 ul collagen 0, 0, 0, (2) Bone (1 of 4) 0.5 20 100 ul
collagen 0, 0, (3), 0 Bone (1 of 4) 0.75 0 100 ul collagen 0, (1),
0, 0 Bone (1 of 4) 0.25 25 100 ul collagen (1), 5, 0, 5 Bone (3 of
4) 2.75 30 100 ul collagen x, 4, 5, 5 Bone (3 of 4) 3.5 35 100 ul
collagen 3, 5, 4, 5 Bone (4 of 4) 4.25 Evero- 0 100 ul collagen 0,
0, 0, 0 No bone (0 of 4) 0 limus 10 100 ul collagen 0, 0, 0, 0 No
bone (0 of 4) 0 15 100 ul collagen 5, 0, (2), 3 Bone (3 of 4) 2.5
20 100 ul collagen 4, 3, 4, 4 Bone (4 of 4) 3.75 0 100 ul collagen
0, 0, 0, 0 No bone (0 of 4) 0 25 100 ul collagen 5, 5, 5, 4 Bone (4
of 4) 4.75 30 100 ul collagen 5, 5, x, 5 Bone (3 of 4) 3.75 35 100
ul collagen 4, 5, 5, 5 Bone (4 of 4) 4.75 Tacro- 0 100 ul collagen
0, 0, 0, 0 No bone (0 of 4) 0 limus 10 100 ul collagen 0, 0, 0, 0
No bone (0 of 4) 0 25 100 ul collagen 4, 5, 4, 4 Bone (4 of 4) 4.25
35 100 ul collagen 2, 5, 4, 5 Bone (4 of 4) 4 0 100 ul collagen 0,
0, 0, 0 No bone (0 of 4) 0 15 100 ul collagen 0, 2, 0, 0 Bone (1 of
4) 0.5 20 100 ul collagen 0, 5, 6, 2 Bone (3 of 4) 3.25 30 100 ul
collagen 0, 3, 5, 2 Bone (3 of 4) 2.5
Sequence CWU 1
1
51396PRTHomo sapiens 1Met Val Ala Gly Thr Arg Cys Leu Leu Ala Leu
Leu Leu Pro Gln Val 1 5 10 15 Leu Leu Gly Gly Ala Ala Gly Leu Val
Pro Glu Leu Gly Arg Arg Lys 20 25 30 Phe Ala Ala Ala Ser Ser Gly
Arg Pro Ser Ser Gln Pro Ser Asp Glu 35 40 45 Val Leu Ser Glu Phe
Glu Leu Arg Leu Leu Ser Met Phe Gly Leu Lys 50 55 60 Gln Arg Pro
Thr Pro Ser Arg Asp Ala Val Val Pro Pro Tyr Met Leu 65 70 75 80 Asp
Leu Tyr Arg Arg His Ser Gly Gln Pro Gly Ser Pro Ala Pro Asp 85 90
95 His Arg Leu Glu Arg Ala Ala Ser Arg Ala Asn Thr Val Arg Ser Phe
100 105 110 His His Glu Glu Ser Leu Glu Glu Leu Pro Glu Thr Ser Gly
Lys Thr 115 120 125 Thr Arg Arg Phe Phe Phe Asn Leu Ser Ser Ile Pro
Thr Glu Glu Phe 130 135 140 Ile Thr Ser Ala Glu Leu Gln Val Phe Arg
Glu Gln Met Gln Asp Ala 145 150 155 160 Leu Gly Asn Asn Ser Ser Phe
His His Arg Ile Asn Ile Tyr Glu Ile 165 170 175 Ile Lys Pro Ala Thr
Ala Asn Ser Lys Phe Pro Val Thr Arg Leu Leu 180 185 190 Asp Thr Arg
Leu Val Asn Gln Asn Ala Ser Arg Trp Glu Ser Phe Asp 195 200 205 Val
Thr Pro Ala Val Met Arg Trp Thr Ala Gln Gly His Ala Asn His 210 215
220 Gly Phe Val Val Glu Val Ala His Leu Glu Glu Lys Gln Gly Val Ser
225 230 235 240 Lys Arg His Val Arg Ile Ser Arg Ser Leu His Gln Asp
Glu His Ser 245 250 255 Trp Ser Gln Ile Arg Pro Leu Leu Val Thr Phe
Gly His Asp Gly Lys 260 265 270 Gly His Pro Leu His Lys Arg Glu Lys
Arg Gln Ala Lys His Lys Gln 275 280 285 Arg Lys Arg Leu Lys Ser Ser
Cys Lys Arg His Pro Leu Tyr Val Asp 290 295 300 Phe Ser Asp Val Gly
Trp Asn Asp Trp Ile Val Ala Pro Pro Gly Tyr 305 310 315 320 His Ala
Phe Tyr Cys His Gly Glu Cys Pro Phe Pro Leu Ala Asp His 325 330 335
Leu Asn Ser Thr Asn His Ala Ile Val Gln Thr Leu Val Asn Ser Val 340
345 350 Asn Ser Lys Ile Pro Lys Ala Cys Cys Val Pro Thr Glu Leu Ser
Ala 355 360 365 Ile Ser Met Leu Tyr Leu Asp Glu Asn Glu Lys Val Val
Leu Lys Asn 370 375 380 Tyr Gln Asp Met Val Val Glu Gly Cys Gly Cys
Arg 385 390 395 223PRTHomo sapiens 2Lys Ile Pro Lys Ala Ser Ser Val
Pro Thr Glu Leu Ser Ala Ile Ser 1 5 10 15 Thr Leu Tyr Leu Asp Asp
Asp 20 328PRTHomo sapiens 3Cys Cys Cys Cys Asp Asp Asp Ser Lys Ile
Pro Lys Ala Ser Ser Val 1 5 10 15 Pro Thr Glu Leu Ser Ala Ile Ser
Thr Leu Tyr Leu 20 25 429PRTHomo sapiensMOD_RES(1)..(1)Where N is
ACETYLATED 4Asn Cys Cys Cys Cys Gly Gly Gly Ser Lys Ile Pro Lys Ala
Ser Ser 1 5 10 15 Val Pro Thr Glu Leu Ser Ala Ile Ser Thr Leu Tyr
Leu 20 25 5431PRTHomo sapiens 5Met His Val Arg Ser Leu Arg Ala Ala
Ala Pro His Ser Phe Val Ala 1 5 10 15 Leu Trp Ala Pro Leu Phe Leu
Leu Arg Ser Ala Leu Ala Asp Phe Ser 20 25 30 Leu Asp Asn Glu Val
His Ser Ser Phe Ile His Arg Arg Leu Arg Ser 35 40 45 Gln Glu Arg
Arg Glu Met Gln Arg Glu Ile Leu Ser Ile Leu Gly Leu 50 55 60 Pro
His Arg Pro Arg Pro His Leu Gln Gly Lys His Asn Ser Ala Pro 65 70
75 80 Met Phe Met Leu Asp Leu Tyr Asn Ala Met Ala Val Glu Glu Gly
Gly 85 90 95 Gly Pro Gly Gly Gln Gly Phe Ser Tyr Pro Tyr Lys Ala
Val Phe Ser 100 105 110 Thr Gln Gly Pro Pro Leu Ala Ser Leu Gln Asp
Ser His Phe Leu Thr 115 120 125 Asp Ala Asp Met Val Met Ser Phe Val
Asn Leu Val Glu His Asp Lys 130 135 140 Glu Phe Phe His Pro Arg Tyr
His His Arg Glu Phe Arg Phe Asp Leu 145 150 155 160 Ser Lys Ile Pro
Glu Gly Glu Ala Val Thr Ala Ala Glu Phe Arg Ile 165 170 175 Tyr Lys
Asp Tyr Ile Arg Glu Arg Phe Asp Asn Glu Thr Phe Arg Ile 180 185 190
Ser Val Tyr Gln Val Leu Gln Glu His Leu Gly Arg Glu Ser Asp Leu 195
200 205 Phe Leu Leu Asp Ser Arg Thr Leu Trp Ala Ser Glu Glu Gly Trp
Leu 210 215 220 Val Phe Asp Ile Thr Ala Thr Ser Asn His Trp Val Val
Asn Pro Arg 225 230 235 240 His Asn Leu Gly Leu Gln Leu Ser Val Glu
Thr Leu Asp Gly Gln Ser 245 250 255 Ile Asn Pro Lys Leu Ala Gly Leu
Ile Gly Arg His Gly Pro Gln Asn 260 265 270 Lys Gln Pro Phe Met Val
Ala Phe Phe Lys Ala Thr Glu Val His Phe 275 280 285 Arg Ser Ile Arg
Ser Thr Gly Ser Lys Gln Arg Ser Gln Asn Arg Ser 290 295 300 Lys Thr
Pro Lys Asn Gln Glu Ala Leu Arg Met Ala Asn Val Ala Glu 305 310 315
320 Asn Ser Ser Ser Asp Gln Arg Gln Ala Cys Lys Lys His Glu Leu Tyr
325 330 335 Val Ser Phe Arg Asp Leu Gly Trp Gln Asp Trp Ile Ile Ala
Pro Glu 340 345 350 Gly Tyr Ala Ala Tyr Tyr Cys Glu Gly Glu Cys Ala
Phe Pro Leu Asn 355 360 365 Ser Tyr Met Asn Ala Thr Asn His Ala Ile
Val Gln Thr Leu Val His 370 375 380 Phe Ile Asn Pro Glu Thr Val Pro
Lys Pro Cys Cys Ala Pro Thr Gln 385 390 395 400 Leu Asn Ala Ile Ser
Val Leu Tyr Phe Asp Asp Ser Ser Asn Val Ile 405 410 415 Leu Lys Lys
Tyr Arg Asn Asn Val Val Arg Ala Cys Gly Cys His 420 425 430
* * * * *