U.S. patent application number 13/839299 was filed with the patent office on 2013-09-26 for macrolide polymorphs, compositions comprising such polymorphs, and methods of use and manufacture thereof.
This patent application is currently assigned to Optimer Pharmaceuticals, Inc.. The applicant listed for this patent is OPTIMER PHARMACEUTICALS, INC.. Invention is credited to Tessie Mary Che, Yu-Hung Chiu, Yoshi Ichikawa, Alex Romero, Youe-Kong Shue.
Application Number | 20130252914 13/839299 |
Document ID | / |
Family ID | 38661942 |
Filed Date | 2013-09-26 |
United States Patent
Application |
20130252914 |
Kind Code |
A1 |
Chiu; Yu-Hung ; et
al. |
September 26, 2013 |
Macrolide Polymorphs, Compositions Comprising Such Polymorphs, and
Methods of Use and Manufacture Thereof
Abstract
The invention relates to novel forms of compounds displaying
broad spectrum antibiotic activity, especially crystalline
polymorphic forms and amorphous forms of such compounds,
compositions comprising such crystalline polymorphic forms and
amorphous forms of such compounds, processes for manufacture and
use thereof. The compounds and compositions of the invention are
useful in the pharmaceutical industry, for example, in the
treatment or prevention of diseases or disorders associated with
the use of antibiotics, chemotherapies, or antiviral therapies,
including, but not limited to, colitis, for example,
pseudo-membranous colitis; antibiotic associated diarrhea; and
infections due to Clostridium difficile ("C. difficile"),
Clostridium perfringens ("C. perfringens"), Staphylococcus species,
for example, methicillin-resistant Staphylococcus, or Enterococcus
including Vancomycin-resistant enterococci.
Inventors: |
Chiu; Yu-Hung; (San Diego,
CA) ; Che; Tessie Mary; (San Diego, CA) ;
Romero; Alex; (Oakland, CA) ; Ichikawa; Yoshi;
(San Diego, CA) ; Shue; Youe-Kong; (Taipei,
TW) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
OPTIMER PHARMACEUTICALS, INC. |
Jersey City |
NJ |
US |
|
|
Assignee: |
Optimer Pharmaceuticals,
Inc.
Jersey City
NJ
|
Family ID: |
38661942 |
Appl. No.: |
13/839299 |
Filed: |
March 15, 2013 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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12397686 |
Mar 4, 2009 |
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13839299 |
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|
12101552 |
Apr 11, 2008 |
7863249 |
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12397686 |
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11831886 |
Jul 31, 2007 |
7378508 |
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12101552 |
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PCT/US2005/002887 |
Jan 31, 2005 |
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11831886 |
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Current U.S.
Class: |
514/28 ;
536/7.1 |
Current CPC
Class: |
C07H 17/08 20130101;
C07D 407/14 20130101; A61P 31/04 20180101; A61K 31/351
20130101 |
Class at
Publication: |
514/28 ;
536/7.1 |
International
Class: |
C07H 17/08 20060101
C07H017/08 |
Claims
1. A crystalline polymorph of tiacumicin comprising a powder x-ray
diffraction pattern with at least peaks at diffraction angles
2.theta. of 7.7.degree., 15.0.degree., and 18.8.degree..+-.0.2.
2. The crystalline polymorph of claim 1, comprising a polymorph
having the chemical structure of Formula I: ##STR00010##
3. The crystalline polymorph of claim 2, further comprising at
least one compound selected from a mixture of tiacumicins.
4. The crystalline polymorph of claim 2, wherein the polymorph of
Formula I is present in at least about 75% to about 99.99%.
5. The crystalline polymorph of claim 4, wherein the polymorph of
Formula I is present in at least about 85%.
6. The crystalline polymorph of claim 4, wherein the polymorph of
Formula I is present in at least about 90%.
7. The crystalline polymorph of claim 4, wherein the polymorph of
Formula I is present in at least about 95%.
8. The crystalline polymorph of claim 4, wherein the polymorph of
Formula I is present in at least about 99%.
9. A crystalline polymorph of tiacumicin comprising: (i) a DSC
endotherm in the range of about 174.degree. C. to about 186.degree.
C.; and (ii) a chemical structure of Formula I: ##STR00011##
10. The crystalline polymorph of claim 9, further comprising at
least one compound selected from a mixture of tiacumicins.
11. The crystalline polymorph of claim 9, wherein the polymorph of
Formula I is present from about 75% to about 99.99%.
12. The crystalline polymorph of claim 9, wherein the polymorph of
Formula I is present in at least about 85%.
13. The crystalline polymorph of claim 11, wherein the polymorph of
Formula I is present in at least about 90%.
14. The crystalline polymorph of claim 11, wherein the polymorph of
Formula I is present in at least about 95%.
15. The crystalline polymorph of claim 11, wherein the polymorph of
Formula I is present in at least about 99%.
16. A crystalline polymorph of tiacumicin comprising: (i) a powder
x-ray diffraction pattern with at least peaks at diffraction angles
2.theta. of 7.7.degree., 15.0.degree., and 18.8.degree..+-.0.2;
(ii) a DSC endotherm in the range of about 174.degree. C. to about
186.degree. C.; and (iii) a chemical structure of Formula I:
##STR00012##
17. The crystalline polymorph of claim 16, further comprising at
least one compound selected from a mixture of tiacumicins.
18. The crystalline polymorph of claim 16, wherein the Compound of
Formula I is present from about 75% to about 99.99%.
19. The crystalline polymorph of claim 16, wherein the Compound of
Formula I is present in about 90%.
20. A composition comprising the crystalline polymorph of claim 1
and a pharmaceutically acceptable carrier.
Description
1. RELATED APPLICATIONS
[0001] The present application is a continuation-in-part
application of PCT Application PCT/US05/02887, filed Jan. 31, 2005,
and claims the benefit of U.S. provisional patent application No.
60/881,950, filed Jan. 22, 2007, the entire disclosures of each are
herein incorporated by reference.
2. FIELD OF THE INVENTION
[0002] The invention encompasses novel forms of compounds
displaying broad spectrum antibiotic activity, especially
crystalline polymorphic forms and amorphous forms of such
compounds, compositions comprising such crystalline polymorphic
forms and amorphous forms of such compounds, processes for
manufacture and use thereof. The compounds and compositions of the
invention are useful in the medical and pharmaceutical industry,
for example, in the treatment or prevention of diseases or
disorders associated with the use of antibiotics, chemotherapies,
or antiviral therapies, including, but not limited to, colitis, for
example, pseudo-membranous colitis; antibiotic associated diarrhea;
and infections due to Clostridium difficile ("C. difficile"),
Clostridium perfringens ("C. perfringens"), Staphylococcus species,
for example, methicillin-resistant Staphylococcus, or Enterococcus
including Vancomycin-resistant enterococci.
3. BACKGROUND OF THE INVENTION
[0003] Antibiotic-associated diarrhea ("AAD") diseases are caused
by toxin producing strains of C. difficile, Staphylococcus aureus
("S. aureus") including methicillin-resistant Staphylococcus aureus
("MRSA") and C. perfringens. AAD represents a major economic burden
to the healthcare system that is conservatively estimated at $3-6
billion per year in excess hospital costs in the United States
alone.
[0004] AAD is a significant problem in hospitals and long-term care
facilities. C. difficile is the leading cause of AAD in the
hospital setting, accounting for approximately 20% of cases of AAD
and the majority of cases of antibiotic-associated colitis ("AAC").
The rising incidence of C. difficile associated diarrhea ("CDAD")
has been attributed to the frequent prescribing of broad-spectrum
antibiotics to hospitalized patients.
[0005] The tiacumicins are a group of 18-membered macrolide
antibiotics originally isolated from the fermentation broth of
Dactylosporangium aurantiacum. The tiacumicins are effective
Gram-positive antibiotics. In particular, tiacumicins, specifically
Tiacumicin B, show activity against a variety of bacterial
pathogens and in particular against C. difficile, a Gram-positive
bacterium (Antimicrob. Agents Chemother., 1991, 1108-1111). A
purification of tiacumicins was carried out in suitable solvents,
wherein tiacumicin B exhibited a melting point of 143-145.degree.
C. (See, e.g., J. E. Hochlowski, et al., J. Antibiotics, vol. XL,
no. 5, pages 575-588 (1987)).
[0006] The polymorphic behavior of a compound can be of crucial
importance in pharmacy and pharmacology. Polymorphs are, by
definition, crystals of the same molecule having different physical
properties as a result of the order of the molecules in the crystal
lattice. The differences in physical properties exhibited by
polymorphs affect pharmaceutical parameters such as storage
stability, compressibility and density (important in formulation
and product manufacturing), and dissolution rates (an important
factor in determining bio-availability). Differences in stability
can result from changes in chemical reactivity (e.g., differential
oxidation, such that a dosage form discolors more rapidly when
comprised of one polymorph than when comprised of another
polymorph) or mechanical changes (e.g., tablets crumble on storage
as a kinetically favored polymorph converts to thermodynamically
more stable polymorph) or both (e.g., tablets of one polymorph are
more susceptible to breakdown at high humidity). As a result of
solubility/dissolution differences, in the extreme case, some
polymorphic transitions may result in lack of potency or, at the
other extreme, toxicity. In addition, the physical properties of a
crystal may be important in processing: for example, one polymorph
might be more likely to form solvates or might be difficult to
filter and wash free of impurities (i.e., particle shape and size
distribution might be different between one polymorph relative to
the other).
[0007] Each pharmaceutical compound has an optimal therapeutic
blood concentration and a lethal concentration. The
bio-availability of the compound determines the dosage strength in
the drug formulation necessary to obtain the ideal blood level. If
the drug can crystallize as two or more polymorphs differing in
bio-availability, the optimal dose will depend on the polymorph
present in the formulation. Some drugs show a narrow margin between
therapeutic and lethal concentrations. Thus, it becomes important
for both medical and commercial reasons to produce and market the
drug in its most thermodynamically stable polymorph, substantially
free of other kinetically favored or disfavored polymorphs.
[0008] Thus, there is a clear need to develop safe and effective
polymorphs of drugs that are efficacious at treating or preventing
disorders associated with bacterial pathogens. The present
inventors have identified novel crystalline and amorphous forms of
18-membered macrolide compounds that exhibit broad spectrum
antibiotic activity.
4. SUMMARY OF THE INVENTION
[0009] The invention encompasses novel crystalline and amorphous
forms of the macrolide compounds that are useful in treating or
preventing bacterial infections and protozoal infections. In an
illustrative embodiment, the novel crystalline and amorphous forms
of the macrolide compounds of the invention exhibit broad spectrum
antibiotic activity. Thus, surprisingly novel crystalline and
amorphous forms of the macrolide compounds have been identified,
which act as antibiotics possessing a broad spectrum of activity in
treating or preventing bacterial infections and protozoal
infections, especially those associated with Gram-positive and
Gram-negative bacteria and in particular, Gram-positive
bacteria.
[0010] In one embodiment, the invention encompasses novel
crystalline and amorphous forms of the macrolide of Formula I:
##STR00001##
[0011] In another embodiment, the invention encompasses a mixture
of compounds with varying amounts of the Compound of Formula I,
which forms have the requisite stability for use in preparing
pharmaceutical compositions.
[0012] In another embodiment, the invention encompasses a polymorph
obtained from a mixture of tiacumicins and a Compound of Formula
I.
[0013] In still another embodiment, the invention encompasses novel
crystalline and amorphous forms of the Compound of Formula I.
[0014] In another embodiment, the invention encompasses a
pharmaceutical composition comprising a Compound of Formula I.
[0015] In another embodiment, the invention encompasses a
pharmaceutical composition comprising a Compound of Formula I,
wherein the Compound of Formula I is present in an amount greater
than 90% by weight.
[0016] In another embodiment, the invention encompasses a
pharmaceutical composition comprising one or more novel crystalline
and amorphous forms of a Compound of Formula I.
[0017] In another embodiment, the invention encompasses a
pharmaceutical composition comprising a mixture of tiacumicins and
Compound of Formula I.
[0018] In another embodiment, the invention encompasses a
pharmaceutical composition comprising a mixture of tiacumicins and
at least about 75% or more by weight of Compound of Formula I. In
another embodiment, the invention encompasses a pharmaceutical
composition comprising a mixture of tiacumicins and at least about
80% or more by weight of Compound of Formula I. In another
embodiment, the invention encompasses a pharmaceutical composition
comprising a mixture of tiacumicins and at least about 85% or more
by weight of Compound of Formula I. In another embodiment, the
invention encompasses a pharmaceutical composition comprising a
mixture of tiacumicins and at least about 90% or more by weight of
Compound of Formula I. In another embodiment, the invention
encompasses a pharmaceutical composition comprising a mixture of
tiacumicins and at least about 95% or more by weight of Compound of
Formula I. In another embodiment, the invention encompasses a
pharmaceutical composition comprising a mixture of tiacumicins and
at least about 99% or more by weight of Compound of Formula I.
[0019] The invention also encompasses methods for treating or
preventing a disease or disorder including, but not limited to,
bacterial infections and protozoal infections comprising
administering to a subject, preferably a mammal, in need thereof a
therapeutically or prophylactically effective amount of a
composition or formulation comprising a compound of the
invention.
[0020] In one illustrative embodiment, the composition or
formulation comprises a mixture of compounds with varying amounts
of the Compound of Formula I. In another embodiment, the
composition or formulation comprises a mixture of tiacumicins and a
Compound of Formula I. In still another embodiment, the composition
or formulation comprises novel crystalline and amorphous forms of
the Compound of Formula I. In still another embodiment, the
composition or formulation comprises novel crystalline and
amorphous forms of the Compound of Formula I and a mixture of
tiacumicins.
[0021] In another particular embodiment, the disease or disorder to
be treated or prevented are caused by toxin producing strains of C.
difficile, Staphylococcus aureus ("S. aureus") including
methicillin-resistant Staphylococcus aureus ("MRSA") and C.
perfringens. In another particular embodiment, the disease or
disorder to be treated or prevented is antibiotic-associated
diarrhea.
5. BRIEF DESCRIPTION OF THE DRAWINGS
[0022] FIG. 1 shows the X-ray powder diffraction patterns of a
first polymorph Compound of Formula I produced from methanol and
water.
[0023] FIG. 2 shows the X-ray powder diffraction patterns of a
second polymorph Compound of Formula I produced from ethyl
acetate.
[0024] FIG. 3 shows the effect of temperature on a mixture of
tiacumicins produced from methanol and water. The DSC indicates an
endothermic curve beginning at 169.degree. C., and weight loss
beginning at 223.degree. C. The endothermic curve at about
177.degree. C. corresponds to the melting of a first polymorph of a
Compound of Formula I.
6. DETAILED DESCRIPTION OF THE DRAWINGS
6.1. General Description
[0025] The invention broadly encompasses mixtures of compounds with
varying amounts of the Compound of Formula I. The inventors have
surprisingly determined that the formation of crystalline
polymorphic forms and amorphous forms of a Compound of Formula I
and optionally mixtures of tiacumicin depends on the selection of
the crystallization solvent and on the method and conditions of
crystallization or precipitation.
[0026] In one embodiment the invention encompasses a mixture of
tiacumicins and a Compound of Formula I. In another embodiment, the
invention encompasses novel crystalline and amorphous forms of the
Compound of Formula I and optionally a mixture of tiacumicins. In
still another embodiment, the invention encompasses novel
crystalline and amorphous forms of the Compound of Formula I and a
mixture of tiacumicins. In another embodiment, the invention
encompasses a mixture of comprising a first polymorph of a Compound
of Formula I, a second polymorph of a Compound of Formula I, and
other polymorphic forms, amorphous forms and mixtures thereof.
[0027] In another particular embodiment, the crystalline polymorphs
and amorphous forms are obtained from a mixture of tiacumicins.
[0028] In another embodiment, a crystalline polymorph of a Compound
of Formula I exhibits a representative powder diffraction pattern
comprising at least peaks at the following diffraction angles
2.theta. of 7.7.degree., 15.0.degree., and 18.8.degree..+-.0.04,
preferably .+-.0.1, more preferably .+-.0.15, even more preferably
.+-.0.2. In another embodiment, a crystalline polymorph of a
Compound of Formula I exhibits a representative powder diffraction
pattern comprising at least peaks at the following diffraction
angles 2.theta. of 7.8.degree., 15.1.degree., and
18.8.degree..+-.0.04, preferably .+-.0.1, more preferably .+-.0.15,
even more preferably .+-.0.2.
[0029] In another embodiment, the polymorph has the chemical
structure:
##STR00002##
[0030] In another embodiment, the polymorph has the chemical
structure of a Compound of Formula I:
##STR00003##
[0031] In another embodiment, the polymorph further comprises at
least one compound selected from a mixture of tiacumicins.
[0032] In another embodiment, the polymorph of Formula I is present
in an amount from at least about 75% to about 99.99%.
[0033] In another embodiment, the polymorph of Formula I is present
in an amount of at least about 75%.
[0034] In another embodiment, the polymorph of Formula I is present
in an amount of at least about 80%.
[0035] In another embodiment, the polymorph of Formula I is present
in an amount of at least about 85%.
[0036] In another embodiment, the polymorph of Formula I is present
in an amount of at least about 90%.
[0037] In another embodiment, the polymorph of Formula I is present
in an amount of at least about 93%.
[0038] In another embodiment, the polymorph of Formula I is present
in an amount of at least about 95%.
[0039] In another embodiment, the polymorph of Formula is present
in an amount of at least about 99%.
[0040] In another embodiment, the crystalline polymorph is obtained
from a mixture of tiacumicins that exhibits a melting point of
about 163.degree. C. to about 169.degree. C. In another embodiment,
the crystalline polymorph is obtained from a mixture of tiacumicins
that exhibits a melting point of about 160.degree. C. to about
170.degree. C. In another embodiment, the crystalline polymorph is
obtained from a mixture of tiacumicins that exhibits a melting
point of about 155.degree. C. to about 175.degree. C.
[0041] In another embodiment, the crystalline polymorph is obtained
from a mixture of tiacumicins and exhibits a DSC endotherm in the
range of about 174.degree. C. to about 186.degree. C.; preferably
175-185.degree. C.
[0042] In another embodiment, the crystalline polymorph is obtained
from a mixture of tiacumicins that exhibits a powder diffraction
pattern comprising at least peaks at the following diffraction
angles 2.theta. of 7.7.degree., 15.0.degree., and
18.8.degree..+-.0.04, preferably .+-.0.1, more preferably .+-.0.15,
even more preferably .+-.0.2 and exhibits a melting point of about
163.degree. C. to about 169.degree. C.
[0043] In another embodiment, the crystalline polymorph is obtained
from a mixture of tiacumicins that exhibits a powder diffraction
pattern comprising at least peaks at the following diffraction
angles 2.theta. of 7.7.degree., 15.0.degree., and
18.8.degree..+-.0.04, preferably .+-.0.1, more preferably .+-.0.15,
even more preferably .+-.0.2 and exhibits a melting point of about
160.degree. C. to about 170.degree. C.
[0044] Another embodiment encompasses a crystalline polymorph
obtained from a mixture of tiacumicins that exhibits a powder
diffraction pattern comprising at least peaks at the following
diffraction angles 2.theta. of 7.7.degree., 15.0.degree., and
18.8.degree..+-.0.04, preferably .+-.0.1, more preferably .+-.0.15,
even more preferably .+-.0.2. In a particular embodiment, the
polymorph has the chemical structure of a Compound of Formula I. In
another embodiment, the crystalline polymorph further comprises at
least one compound selected from a mixture of tiacumicins.
[0045] In another embodiment, a crystalline polymorph is obtained
from a mixture of tiacumicins that exhibits a melting point of
about 150.degree. C. to about 156.degree. C.
[0046] In another embodiment, a crystalline polymorph is obtained
from a mixture of tiacumicins that exhibits a powder diffraction
pattern comprising at least peaks at the following diffraction
angles 2.theta. of 7.4.degree., 15.5.degree., and
18.8.degree..+-.0.2 and exhibits a melting point of about
150.degree. C. to about 156.degree. C.
[0047] Another embodiment of the invention encompasses
pharmaceutical compositions comprising a therapeutically or
prophylactically effective amount of a crystalline polymorph of a
Compound of Formula:
##STR00004##
and a pharmaceutically acceptable carrier.
[0048] In a particular embodiment, the pharmaceutical composition
comprises a first polymorph of a Compound of Formula I, a second
polymorph of a Compound of Formula I, other polymorphic forms of a
Compound of Formula I, amorphous forms of a Compound of Formula I,
and mixtures thereof.
[0049] In another embodiment, the crystalline polymorph of the
pharmaceutical composition has peaks at the following diffraction
angles 2.theta. of 7.7.degree., 15.0.degree., and
18.8.degree..+-.0.04, preferably .+-.0.1, more preferably .+-.0.15,
even more preferably .+-.0.2.
[0050] In another embodiment, the crystalline polymorph of the
pharmaceutical composition further comprises at least one compound
selected from a mixture of tiacumicins.
[0051] In another embodiment, the Compound of Formula I is present
from at least about 75% to about 99.99%, preferably about 75%,
about 85%, about 95%, or about 99%.
[0052] In another embodiment, the crystalline polymorph of the
pharmaceutical composition exhibits a melting point of about
163.degree. C. to about 169.degree. C.
[0053] Another embodiment encompasses a pharmaceutical composition
comprising a crystalline polymorph of tiacumicin comprising peaks
at the following diffraction angles 2.theta. of 7.6.degree.,
15.4.degree., and 18.8.degree..+-.0.04, preferably .+-.0.1, more
preferably .+-.0.15, even more preferably .+-.0.2. In a particular
embodiment, the pharmaceutical composition further comprises at
least one compound selected from a mixture of tiacumicins. In
another particular embodiment, the Compound of Formula I is present
from about 75% to about 99.99%, preferably 75%, 85%, 95%, or
99%.
[0054] In another embodiment, the invention encompasses a
pharmaceutical composition containing stereomerically pure
R-Tiacumicin and less than 15% of a mixture of tiacumicins. In
another embodiment, the invention encompasses a pharmaceutical
composition containing stereomerically pure R-Tiacumicin and less
than 10% of a mixture of tiacumicins. In another embodiment, the
invention encompasses a pharmaceutical composition containing
stereomerically pure R-Tiacumicin and less than 7% of a mixture of
tiacumicins. In another embodiment, the invention encompasses a
pharmaceutical composition containing stereomerically pure
R-Tiacumicin and less than 5% of a mixture of tiacumicins. In
another embodiment, the invention encompasses a pharmaceutical
composition containing stereomerically pure R-Tiacumicin and less
than 1% of a mixture of tiacumicins. In another embodiment, the
invention encompasses a pharmaceutical composition containing
stereomerically pure R-Tiacumicin and less than 15% of a mixture of
S-Tiacumicin. In another embodiment, the invention encompasses a
pharmaceutical composition containing stereomerically pure
R-Tiacumicin and less than 10% of a mixture of S-Tiacumicin. In
another embodiment, the invention encompasses a pharmaceutical
composition containing stereomerically pure R-Tiacumicin and less
than 7% of a mixture of S-Tiacumicin. In another embodiment, the
invention encompasses a pharmaceutical composition containing
stereomerically pure R-Tiacumicin and less than 5% of a mixture of
S-Tiacumicin. In another embodiment, the invention encompasses a
pharmaceutical composition containing stereomerically pure
R-Tiacumicin and less than 1% of a mixture of S-Tiacumicin. In
another embodiment, the invention encompasses a pharmaceutical
composition containing stereomerically pure R-Tiacumicin and less
than 15% of a mixture of Lipiarmycin A4. In another embodiment, the
invention encompasses a pharmaceutical composition containing
stereomerically pure R-Tiacumicin and less than 10% of a mixture of
Lipiarmycin A4. In another embodiment, the invention encompasses a
pharmaceutical composition containing stereomerically pure
R-Tiacumicin and less than 7% of a mixture of Lipiarmycin A4. In
another embodiment, the invention encompasses a pharmaceutical
composition containing stereomerically pure R-Tiacumicin and less
than 5% of a mixture of Lipiarmycin A4. In another embodiment, the
invention encompasses a pharmaceutical composition containing
stereomerically pure R-Tiacumicin and less than 1% of a mixture of
Lipiarmycin A4.
[0055] In another embodiment, the crystalline polymorph of the
pharmaceutical composition exhibits a melting point of about
153.degree. C. to about 156.degree. C.
[0056] In another embodiment, the therapeutically or
prophylactically effective amount is from about 0.01 mg/kg to about
1000 mg/kg, preferably 0.01, 0.1, 1, 2.5, 5, 10, 20, 50, 100, 250,
or 500 mg/kg.
[0057] In another embodiment, the crystalline polymorph of the
pharmaceutical composition is suitable for parenteral
administration, preferably intravenous, intramuscular, or
intraarterial.
[0058] In another embodiment, the crystalline polymorph of the
pharmaceutical composition is suitable for peroral
administration.
[0059] Another embodiment of the invention encompasses a method for
treating a bacterial infection comprising administering a
pharmaceutical composition comprising a polymorph of the invention
to a subject in need thereof.
[0060] In a particular embodiment, the bacterial infection is in
the gastrointestinal tract, particularly AAC or AAD.
6.2. Definitions
[0061] The term "antibiotic-associated condition" refers to a
condition resulting when antibiotic therapy disturbs the balance of
the microbial flora of the gut, allowing pathogenic organisms such
as enterotoxin producing strains of C. difficile, S. aureus and C.
perfringens to flourish. These organisms can cause diarrhea,
pseudomembranous colitis, and colitis and are manifested by
diarrhea, urgency, abdominal cramps, tenesmus, and fever among
other symptoms. Diarrhea, when severe, causes dehydration and the
medical complications associated with dehydration.
[0062] The term "asymmetrically substituted" refers to a molecular
structure in which an atom having four tetrahedral valences is
attached to four different atoms or groups. The commonest cases
involve the carbon atom. In such cases, two optical isomers (D- and
L-enantiomers or R- and S-enantiomers) per carbon atom result which
are nonsuperposable mirror images of each other. Many compounds
have more than one asymmetric carbon. This results in the
possibility of many optical isomers, the number being determined by
the formula 2n, where n is the number of asymmetric carbons.
[0063] The term "broth" as used herein refers to the fluid culture
medium as obtained during or after fermentation. Broth comprises a
mixture of water, the desired antibiotic(s), unused nutrients,
living or dead organisms, metabolic products, and the adsorbent
with or without adsorbed product.
[0064] As used herein and unless otherwise indicated, the terms
"bacterial infection(s)" and "protozoal infection(s)" are used
interchangeably and include bacterial infections and protozoal
infections that occur in mammals, fish and birds as well as
disorders related to bacterial infections and protozoal infections
that may be treated or prevented by antibiotics such as the
Compounds of the Invention. Such bacterial infections and protozoal
infections, and disorders related to such infections, include the
following: disorders associated with the use of antibiotics,
chemotherapies, or antiviral therapies, including, but not limited
to, colitis, for example, pseudo-membranous colitis, antibiotic
associated diarrhea, and infections due to Clostridium difficile,
Clostridium perfringens, Staphylococcus species,
methicillin-resistant Staphylococcus, or Enterococcus including
Vancomycin-resistant enterococci; antibiotic-associated diarrhea
including those caused by toxin producing strains of C. difficile,
S. aureus including methicillin-resistant Staphylococcus aureus,
and C. perfringens; and antibiotic-associated colitis; pneumonia,
otitis media, sinusitis, bronchitis, tonsillitis and mastoiditis
related to infection by Staphylococcus pneumoniae, Haemophilus
influenzae, Moraxella catarrhalis, Staphlococcus aureus, or
Peptostreptococcus spp.; pharynigis, rheumatic fever and
glomerulonephritis related to infection by Streptococcus pyogenes,
Groups C and G streptococci, Clostridium diptheriae, or
Actinobacillus haemolyticum; respiratory tract infections related
to infection by Mycoplasma pneumoniae, Legionella pneumophila,
Streptococcus pneumoniae, Haemophilus influenzae, or Chlamydia
pneumoniae; uncomplicated skin and soft tissue infections,
abscesses and osteomyelitis, and puerperal fever related to
infection by Staphlococcus aureus, coagulase-positive staphlococci
(e.g., S. epidermis and S. hemolyticus), Staphylococcus pyogenes,
Streptococcus agalactiae, Streptococcal groups C-F (minute-colony
streptococci), viridans streptococci, Corynebacterium minutissimum,
Clostridium spp., or Bartonella henselae; uncomplicated acute
urinary tract infections related to infection by Staphylococcus
saprophyticus or Enterococcus spp.; urethritis and cervicitis; and
sexually transmitted diseases related to infection by Chlamydia
trachomatis, Haemophilus ducreyi, Treponema pallidum, Ureaplasma
urealyticum, or Neiserria gonorrhea; toxin diseases related to
infection by S. aureus (food poisoning and Toxic Shock Syndrome),
or Groups A, B and C streptococci; ulcers related to infection by
Helicobacter pylori, systemic febrile syndromes related to
infection by Borrelia recurrentis; Lyme disease related to
infection by Borrelia burgdorferi, conjunctivitis, keratitis, and
dacrocystitis related to infection by Chlamydia trachomatis,
Neisseria gonorrhoeae, S. aureus, S. pneumoniae, S. pyogenes, H.
influenzae, or Listeria spp.; disseminated Mycobacterium avium
complex (MAC) disease related to infection by Mycobacterium avium,
or Mycobacterium intracellulare; gastroenteritis related to
infection by Campylobacter jejuni, intestinal protozoa related to
infection by Cryptosporidium spp.; odontogenic infection related to
infection by viridans streptococci; persistent cough related to
infection by Bordetella pertussis; gas gangrene related to
infection by Clostridium perfringens or Bacteroides spp.; and
atherosclerosis related to infection by Helicobacter pylori or
Chlamydia pneumoniae. Bacterial infections and protozoal infections
and disorders related to such infections that may be treated or
prevented in animals include the following: bovine respiratory
disease related to infection by P. haem., P. multocida, Mycoplasma
bovis, or Bordetella spp.; cow enteric disease related to infection
by E. coli or protozoa (e.g., coccidia, cryptosporidia, etc.);
dairy cow mastitis related to infection by Staph. aureus, Strep.
uberis, Strep. agalactiae, Strep. dysgalactiae, Klebsiella spp.,
Corynebacterium, or Enterococcus spp.; swine respiratory disease
related to infection by A. pleuro., P. multocida or Mycoplasma
spp.; swine enteric disease related to infection by E. coli
Lawsonia intracellularis, Salmonella, or Serpulina hyodyisinteriae;
cow foxtrot related to infection by Fusobacterium spp.; cow
metritis related to infection by E. coli; cow hairy warts related
to infection by Fusobacterium necrophorum or Bacteroides nodosus;
cow pink-eye related to infection by Moraxella bovis; cow premature
abortion related to infection by protozoa (e.g., neosporium)
urinary tract infection in dogs and cats related to infection by E.
coli; skin and soft tissue infections in dogs and cats related to
infection by Staph. epidermidis, Staph. intermedius, coagulase neg.
Staph. or P. multocida; and dental or mouth infections in dogs and
cats related to infection by Alcaligenes spp., Bacteroides spp.,
Clostridium spp., Enterobacter spp., Eubacterium,
Peptostreptococcus, Porphyromonas, or Prevotella. Other bacterial
infections and protozoal infections and disorders related to such
infections that may be treated or prevented in accord with the
methods of the invention are referred to in Sanford, J. P., et al.,
"The Sanford Guide To Antimicrobial Therapy," 27.sup.th Edition
(Antimicrobial Therapy, Inc., 1996).
[0065] As used herein and unless otherwise indicated, the term
"binders" refers to agents used to impart cohesive qualities to the
powdered material. Binders, or "granulators" as they are sometimes
known, impart cohesiveness to the tablet formulation, which insures
the tablet remaining intact after compression, as well as improving
the free-flowing qualities by the formulation of granules of
desired hardness and size. Materials commonly used as binders
include starch; gelatin; sugars, such as sucrose, glucose,
dextrose, molasses, and lactose; natural and synthetic gums, such
as acacia, sodium alginate, extract of Irish moss, panwar gum,
ghatti gum, mucilage of isapol husks, carboxymethylcellulose,
methylcellulose, polyvinylpyrrolidone, Veegum, microcrystalline
cellulose, microcrystalline dextrose, amylose, and larch
arabogalactan, and the like.
[0066] As used herein and unless otherwise indicated, the terms
"biohydrolyzable amide," "biohydrolyzable ester," "biohydrolyzable
carbamate," "biohydrolyzable carbonate," "biohydrolyzable ureide,"
"biohydrolyzable phosphate" mean an amide, ester, carbamate,
carbonate, ureide, or phosphate, respectively, of a compound that
either: 1) does not interfere with the biological activity of the
compound but can confer upon that compound advantageous properties
in vivo, such as uptake, duration of action, or onset of action; or
2) is biologically inactive but is converted in vivo to the
biologically active compound. Examples of biohydrolyzable esters
include, but are not limited to, lower alkyl esters, lower
acyloxyalkyl esters (such as acetoxylmethyl, acetoxyethyl,
aminocarbonyloxy-methyl, pivaloyloxymethyl, and pivaloyloxyethyl
esters), lactonyl esters (such as phthalidyl and thiophthalidyl
esters), lower alkoxyacyloxyalkyl esters (such as
methoxycarbonyloxy-methyl, ethoxycarbonyloxyethyl and
isopropoxycarbonyloxyethyl esters), alkoxyalkyl esters, choline
esters, and acylamino alkyl esters (such as acetamidomethyl
esters). Examples of biohydrolyzable amides include, but are not
limited to, lower alkyl amides, a amino acid amides, alkoxyacyl
amides, and alkylaminoalkyl-carbonyl amides. Examples of
biohydrolyzable carbamates include, but are not limited to, lower
alkylamines, substituted ethylenediamines, aminoacids,
hydroxyalkylamines, heterocyclic and heteroaromatic amines, and
polyether amines.
[0067] As used herein and unless otherwise indicated, the term
"carrier" refers to a diluent, adjuvant, excipient, or vehicle with
which a composition is administered. Such pharmaceutical carriers
can be sterile liquids, such as water and oils, including those of
petroleum, animal, vegetable or synthetic origin, such as peanut
oil, soybean oil, mineral oil, sesame oil and the like.
[0068] As used herein and unless otherwise indicated, the term
"Compounds of the Invention" means, collectively, a Compound of
Formula I and/or pharmaceutically acceptable salts and polymorphs
thereof. The compounds of the invention are identified herein by
their chemical structure and/or chemical name. Where a compound is
referred to by both a chemical structure and a chemical name, and
that chemical structure and chemical name conflict, the chemical
structure is determinative of the compound's identity. The
compounds of the invention may contain one or more chiral centers
and/or double bonds and, therefore, exist as stereoisomers, such as
double-bond isomers (i.e., geometric isomers), enantiomers, or
diastereomers. According to the invention, the chemical structures
depicted herein, and therefore the compounds of the invention,
encompass all of the corresponding compound's enantiomers and
stereoisomers, that is, both the stereomerically pure form (e.g.,
geometrically pure, enantiomerically pure, or diastereomerically
pure) and enantiomeric and stereoisomeric mixtures. Enantiomeric
and stereoisomeric mixtures can be resolved into their component
enantiomers or stereoisomers by well known methods, such as
chiral-phase gas chromatography, chiral-phase high performance
liquid chromatography, crystallizing the compound as a chiral salt
complex, or crystallizing the compound in a chiral solvent.
Enantiomers and stereoisomers can also be obtained from
stereomerically- or enantiomerically-pure intermediates, reagents,
and catalysts by well known asymmetric synthetic methods. The
Compounds of the Invention are preferably substantially
stereomerically pure. In a particular embodiment, the term
"Compounds of the Invention" refers to a Compound of Formula that
is greater than 75% pure, preferably greater than 85% pure, more
preferably greater than 95% pure and most preferably greater than
99% pure and polymorphic form (e.g., a polymorph of Compound of
Formula I) and amorphous forms thereof.
[0069] As used herein and unless otherwise indicated, "diluents"
are inert substances added to increase the bulk of the formulation
to make the tablet a practical size for compression. Commonly used
diluents include calcium phosphate, calcium sulfate, lactose,
kaolin, mannitol, sodium chloride, dry starch, powdered sugar,
silica, and the like.
[0070] As used herein and unless otherwise indicated,
"disintegrators" or "disintegrants" are substances that facilitate
the breakup or disintegration of tablets after administration.
Materials serving as disintegrants have been chemically classified
as starches, clays, celluloses, algins, or gums. Other
disintegrators include Veegum HV, methylcellulose, agar, bentonite,
cellulose and wood products, natural sponge, cation-exchange
resins, alginic acid, guar gum, citrus pulp, cross-linked
polyvinylpyrrolidone, carboxymethylcellulose, and the like.
[0071] When administered to a subject (e.g., to an animal for
veterinary use or to a human for clinical use) the compounds of the
invention are administered in isolated form. As used herein and
unless otherwise indicated, "isolated" means that the compounds of
the invention are separated from other components of either (a) a
natural source, such as a plant or cell, preferably bacterial
culture, or (b) a synthetic organic chemical reaction mixture,
preferably, via conventional techniques, the compounds of the
invention are purified. As used herein, "purified" means that when
isolated, the isolate contains at least about 70% preferably at
least about 80%, more preferably at least about 90%, even more
preferably at least about 95%, and most preferably at least about
99% of a compound of the invention by weight of the isolate.
[0072] The term "macrolide" or "macrocycle" refers to organic
molecules with large ring structures usually containing over 10
atoms.
[0073] The term "18-membered macrocycles" refers to organic
molecules with ring structures containing 18 atoms.
[0074] The term "MIC" or "minimum inhibitory concentration" refers
to the lowest concentration of an antibiotic that is needed to
inhibit growth of a bacterial isolate in vitro. A common method for
determining the MIC of an antibiotic is to prepare several tubes
containing serial dilutions of the antibiotic, that are then
inoculated with the bacterial isolate of interest. The MIC of an
antibiotic can be determined from the tube with the lowest
concentration that shows no turbidity (no growth).
[0075] The term "MIC50" refers to the lowest concentration of
antibiotic required to inhibit the growth of 50% of the bacterial
strains tested within a given bacterial species.
[0076] The term "MIC90" refers to the lowest concentration of
antibiotic required to inhibit the growth of 90% of the bacterial
strains tested within a given bacterial species.
[0077] As used herein and unless otherwise indicated, the term
"mixture of tiacumicins" refers to a composition containing at
least one macrolide compound from the family of compounds known
tiacumicins. In another embodiment, the term "mixture of
tiacumicins" includes a mixture containing at least one member of
the compounds known tiacumicins and a Compound of Formula I,
wherein the Compound of Formula I is present in an amount of about
50%, 60%, 70%, 80%, 90%, 95%, 99%, 99.9%, or 99.99% by weight. In
particular, the term "mixture of tiacumicins" refers to a
compositions comprising a Compound of Formula I, wherein the
Compound of Formula I has a relative retention time ("RTT") ratio
of 1.0, and further comprising at least one of the following
compounds:
##STR00005## ##STR00006## ##STR00007## ##STR00008##
[0078] In certain illustrative embodiments, when compound 109 is
present in the mixture optionally one of compounds 110, 111, and/or
112 is also present in the mixture. Compound 109 is also sometimes
referred to as Lipiarmycin A4. Compound 110 is also sometimes
referred to as Tiacumicin F. Compound III is also sometimes
referred to as Tiacumicin C. Compound 112 is also sometimes
referred to as Tiacumicin A.
[0079] As used herein, and unless otherwise indicated, the terms
"optically pure," "stereomerically pure," and "substantially
stereomerically pure" are used interchangeably and mean one
stereoisomer of a compound or a composition that comprises one
stereoisomer of a compound and is substantially free of other
stereoisomer(s) of that compound. For example, a stereomerically
pure compound or composition of a compound having one chiral center
will be substantially free of the opposite enantiomer of the
compound. A stereomerically pure compound or composition of a
compound having two chiral centers will be substantially free of
other diastereomers of the compound. A typical stereomerically pure
compound comprises greater than about 80% by weight of one
stereoisomer of the compound and less than about 20% by weight of
other stereoisomers of the compound, more preferably greater than
about 90% by weight of one stereoisomer of the compound and less
than about 10% by weight of the other stereoisomers of the
compound, even more preferably greater than about 95% by weight of
one stereoisomer of the compound and less than about 5% by weight
of the other stereoisomers of the compound, and most preferably
greater than about 97% by weight of one stereoisomer of the
compound and less than about 3% by weight of the other
stereoisomers of the compound.
[0080] As used herein and unless otherwise indicated,
"pharmaceutically acceptable" refers to materials and compositions
that are physiologically tolerable and do not typically produce an
allergic or similar untoward reaction, such as gastric upset,
dizziness and the like, when administered to a human. Typically, as
used herein, the term "pharmaceutically acceptable" means approved
by a regulatory agency of the Federal or a state government or
listed in the U.S. Pharmacopeia or other generally recognized
pharmacopeia for use in animals, and more particularly in
humans.
[0081] As used herein and unless otherwise indicated, the term
"pharmaceutically acceptable hydrate" means a Compound of the
Invention that further includes a stoichiometric or
non-stoichiometric amount of water bound by non-covalent
intermolecular forces.
[0082] As used herein and unless otherwise indicated, the term
"pharmaceutically acceptable polymorph" refers to a Compound of the
Invention that exists in several distinct forms (e.g., crystalline,
amorphous), the invention encompasses all of these forms.
[0083] As used herein and unless otherwise indicated, the term
"pharmaceutically acceptable prodrug" means a derivative of a
modified polymorph of a compound of Formula I that can hydrolyze,
oxidize, or otherwise react under biological conditions (in vitro
or in vivo) to provide the compound. Examples of prodrugs include,
but are not limited to, compounds that comprise biohydrolyzable
moieties such as biohydrolyzable amides, biohydrolyzable esters,
biohydrolyzable carbamates, biohydrolyzable carbonates,
biohydrolyzable ureides, and biohydrolyzable phosphate analogues.
Other examples of prodrugs include compounds that comprise
oligonucleotides, peptides, lipids, aliphatic and aromatic groups,
or NO, NO.sub.2, ONO, and ONO.sub.2 moieties. Prodrugs can
typically be prepared using well known methods, such as those
described in Burger's Medicinal Chemistry and Drug Discovery, 172
178, 949 982 (Manfred E. Wolff ed., 5th ed. 1995), and Design of
Prodrugs (H. Bundgaard ed., Elselvier, New York 1985).
[0084] The phrase "pharmaceutically acceptable salt(s)," as used
herein includes but is not limited to salts of acidic or basic
groups that may be present in compounds used in the present
compositions. Compounds included in the present compositions that
are basic in nature are capable of forming a wide variety of salts
with various inorganic and organic acids. The acids that may be
used to prepare pharmaceutically acceptable acid addition salts of
such basic compounds are those that form non-toxic acid addition
salts, i.e., salts containing pharmacologically acceptable anions
including, but not limited to, sulfuric, citric, maleic, acetic,
oxalic, hydrochloride, hydrobromide, hydroiodide, nitrate, sulfate,
bisulfate, phosphate, acid phosphate, isonicotinate, acetate,
lactate, salicylate, citrate, acid citrate, tartrate, oleate,
tannate, pantothenate, bitartrate, ascorbate, succinate, maleate,
gentisinate, fumarate, gluconate, glucaronate, saccharate, formate,
benzoate, glutamate, methanesulfonate, ethanesulfonate,
benzenesulfonate, p-toluenesulfonate and pamoate (i.e.,
1,1'-methylene-bis-(2-hydroxy-3-naphthoate)) salts. Compounds
included in the present compositions that include an amino moiety
may form pharmaceutically acceptable salts with various amino
acids, in addition to the acids mentioned above. Compounds,
included in the present compositions, which are acidic in nature
are capable of forming base salts with various pharmacologically
acceptable cations. Examples of such salts include alkali metal or
alkaline earth metal salts and, particularly, calcium, magnesium,
sodium lithium, zinc, potassium, and iron salts.
[0085] As used herein and unless otherwise indicated, the term
"prophylactically effective" refers to an amount of a Compound or
Composition of the Invention or a pharmaceutically acceptable salt,
solvate, polymorph, or prodrug thereof causing a reduction of the
risk of acquiring a given disease or disorder. Accordingly, the
Compounds of the Invention may be used for the prevention of one
disease or disorder and concurrently treating another (e.g.,
prevention of AAC, while treating urinary AAD). In certain
embodiments, the compositions of the invention are administered to
a patient, preferably a human, as a preventative measure against
such diseases. As used herein, "prevention" or "preventing" refers
to a reduction of the risk of acquiring a given disease or
disorder.
[0086] As used herein, the term "subject" can be a mammal,
preferably a human or an animal. The subject being treated is a
patient in need of treatment.
[0087] As used herein and unless otherwise indicated, the phrase
"therapeutically effective amount" of a Compound or Composition of
the Invention or a pharmaceutically acceptable salt, solvate,
polymorph, or prodrug thereof is measured by the therapeutic
effectiveness of a compound of the invention, wherein at least one
adverse effect of a disorder is ameliorated or alleviated. In one
embodiment, the term "therapeutically effective amount" means an
amount of a drug or Compound of the Invention that is sufficient to
provide the desired local or systemic effect and performance at a
reasonable benefit/risk ratio attending any medical treatment. In
one embodiment, the phrase "therapeutically effective amount" of a
composition of the invention is measured by the therapeutic
effectiveness of a compound of the invention to alleviate at least
one symptom associated with bacterial or protazoal infections.
Surprisingly, the inventors have found that therapeutically
effective amounts of the compounds of the invention are useful in
treating or preventing bacterial and protazoal infections.
[0088] As used herein and unless otherwise indicated, the terms
"treatment" or "treating" refer to an amelioration of a disease or
disorder, or at least one discernible symptom thereof, preferably
associated with a bacterial or protozoal infection. In another
embodiment, "treatment" or "treating" refers to an amelioration of
at least one measurable physical parameter, not necessarily
discernible by the patient. In yet another embodiment, "treatment"
or "treating" refers to inhibiting the progression of a disease or
disorder, either physically, e.g., stabilization of a discernible
symptom, physiologically, for example, stabilization of a physical
parameter, or both. In yet another embodiment, "treatment" or
"treating" refers to delaying the onset of a disease or
disorder.
6.3. Compositions of the Invention for Therapeutic/Prophylactic
Administration
[0089] The invention encompasses compositions comprising a first
polymorph of a Compound of Formula I, a second polymorph of a
Compound of Formula I, other polymorphic forms, amorphous form or
mixtures thereof of a mixture of tiacumicins with varying amounts
of the Compound of Formula I.
[0090] The invention further encompasses an antibiotic composition
that is a mixture of tiacumicins for use in treating CDAD as well
as, AAD and AAC. The mixture of tiacumicins contains about 76 to
about 100% of a Compound of Formula I, which belongs to the
tiacumicin family of 18-member macrolide.
[0091] Due to the activity of the Compounds of the Invention, the
compounds are advantageously useful in veterinary and human
medicine. The Compounds of the Invention are useful for the
treatment or prevention of bacterial and protozoal infections. In
some embodiments, the subject has an infection but does not exhibit
or manifest any physiological symptoms associated with an
infection.
[0092] The invention provides methods of treatment and prophylaxis
by administration to a patient of a therapeutically effective
amount of a composition comprising a crystalline polymorph or
amorphous form of a Compound of the Invention. The patient is a
mammal, including, but not limited, to an animal such a cow, horse,
sheep, pig, chicken, turkey, quail, cat, dog, mouse, rat, rabbit,
guinea pig, etc., and is more preferably a human.
[0093] The present compositions, which comprise one or more
crystalline polymorph or amorphous form of a Compounds of the
Invention or a mixture of tiacumicins may be administered by any
convenient route, for example, peroral administration, parenteral
administration, by infusion or bolus injection, by absorption
through epithelial or mucocutaneous linings (e.g., oral mucosa,
rectal and intestinal mucosa, etc.) and may be administered
together with another biologically active agent. Administration can
be systemic or local. Various delivery systems are known, e.g.,
encapsulation in liposomes, microparticles, microcapsules,
capsules, etc., and can be used to administer a compound of the
invention. In certain embodiments, more than one Compound of the
Invention and mixture of tiacumicins is administered to a patient.
Methods of administration include but are not limited to
intradermal, intramuscular, intraperitoneal, intravenous,
subcutaneous, intranasal, epidural, oral, sublingual, intranasal,
intracerebral, intravaginal, transdermal, rectally, by inhalation,
or topically, particularly to the ears, nose, eyes, or skin. The
preferred mode of administration is left to the discretion of the
practitioner, and will depend in-part upon the site of the medical
condition. In most instances, administration will result in the
release of the crystalline polymorph or amorphous form of a
Compound of the Invention into the bloodstream.
[0094] In specific embodiments, it may be desirable to administer
one or more crystalline polymorph or amorphous form of a Compound
of the Invention locally to the area in need of treatment. This may
be achieved, for example, and not by way of limitation, by local
infusion during surgery, topical application, e.g., in conjunction
with a wound dressing after surgery, by injection, by means of a
catheter, by means of a suppository, or by means of an implant,
said implant being of a porous, non-porous, or gelatinous material,
including membranes, such as sialastic membranes, or fibers. In one
embodiment, administration can be by direct injection at the site
(or former site) of an atherosclerotic plaque tissue.
[0095] Pulmonary administration can also be employed, e.g., by use
of an inhaler or nebulizer, and formulation with an aerosolizing
agent, or via perfusion in a fluorocarbon or synthetic pulmonary
surfactant. In certain embodiments, the compounds of the invention
can be formulated as a suppository, with traditional binders and
vehicles such as triglycerides.
[0096] In another embodiment, the a crystalline polymorph or
amorphous form of a Compound of the Invention can be delivered in a
vesicle, in particular a liposome (see Langer, 1990, Science
249:1527-1533; Treat et al., in Liposomes in the Therapy of
Infectious Disease and Cancer, Lopez-Berestein and Fidler (eds.),
Liss, New York, pp. 353-365 (1989); Lopez-Berestein, ibid., pp.
317-327; see generally ibid.).
[0097] In yet another embodiment, the compounds of the invention
can be delivered in a controlled release system. In one embodiment,
a pump may be used (see Langer, supra; Sefton, 1987, CRC Crit. Ref
Biomed. Eng. 14:201; Buchwald et al., 1980, Surgery 88:507 Saudek
et al., 1989, N. Engl. J. Med. 321:574). In another embodiment,
polymeric materials can be used (see Medical Applications of
Controlled Release, Langer and Wise (eds.), CRC Pres., Boca Raton,
Fla. (1974); Controlled Drug Bioavailability, Drug Product Design
and Performance, Smolen and Ball (eds.), Wiley, New York (1984);
Ranger and Peppas, 1983, J. Macromol. Sci. Rev. Macromol. Chem.
23:61; see also Levy et al., 1985, Science 228:190; During et al.,
1989, Ann. Neurol. 25:351; Howard et al., 1989, J. Neurosurg.
71:105). In yet another embodiment, a controlled-release system can
be placed in proximity of the target of the compounds of the
invention, e.g., the liver, thus requiring only a fraction of the
systemic dose (see, e.g., Goodson, in Medical Applications of
Controlled Release, supra, vol. 2, pp. 115-138 (1984)). Other
controlled-release systems discussed in the review by Langer, 1990,
Science 249:1527-1533) may be used.
[0098] The present compositions will contain a therapeutically
effective amount of a crystalline polymorph or amorphous form of a
Compound of the Invention, optionally more than one crystalline
polymorph or amorphous form of a Compound of the Invention,
preferably in purified form, together with a suitable amount of a
pharmaceutically acceptable vehicle so as to provide the form for
proper administration to the patient.
[0099] In a specific embodiment, the term "pharmaceutically
acceptable" means approved by a regulatory agency of the Federal or
a state government or listed in the U.S. Pharmacopeia or other
generally recognized pharmacopeia for use in animals, and more
particularly in humans. The term "vehicle" refers to a diluent,
adjuvant, excipient, or carrier with which a compound of the
invention is administered. Such pharmaceutical vehicles can be
liquids, such as water and oils, including those of petroleum,
animal, vegetable or synthetic origin, such as peanut oil, soybean
oil, mineral oil, sesame oil and the like. The pharmaceutical
vehicles can be saline, gum acacia, gelatin, starch paste, talc,
keratin, colloidal silica, urea, and the like. In addition,
auxiliary, stabilizing, thickening, lubricating and coloring agents
may be used. When administered to a patient, the compounds of the
invention and pharmaceutically acceptable vehicles are preferably
sterile. Water is a preferred vehicle when the compound of the
invention is administered intravenously. Saline solutions and
aqueous dextrose and glycerol solutions can also be employed as
liquid vehicles, particularly for injectable solutions. Suitable
pharmaceutical vehicles also include excipients such as starch,
glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk,
silica gel, sodium stearate, glycerol monostearate, talc, sodium
chloride, dried skim milk, glycerol, propylene, glycol, water,
ethanol and the like. The present compositions, if desired, can
also contain minor amounts of wetting or emulsifying agents, or pH
buffering agents.
[0100] The present compositions can take the form of solutions,
suspensions, emulsion, tablets, pills, pellets, capsules, capsules
containing liquids, powders, sustained-release formulations,
suppositories, emulsions, aerosols, sprays, suspensions, or any
other form suitable for use. In one embodiment, the
pharmaceutically acceptable vehicle is a capsule (see e.g., U.S.
Pat. No. 5,698,155). Other examples of suitable pharmaceutical
vehicles are described in "Remington's Pharmaceutical Sciences" by
A. R. Gennaro.
[0101] In a preferred embodiment, the crystalline polymorph or
amorphous form of a Compound of the Invention is formulated in
accordance with routine procedures as a pharmaceutical composition
adapted for intravenous administration to human beings. Typically,
a crystalline polymorph or amorphous form of a Compound of the
Invention for intravenous administration is a solution in sterile
isotonic aqueous buffer. Where necessary, the compositions may also
include a solubilizing agent. Compositions for intravenous
administration may optionally include a local anesthetic such as
lidocaine to ease pain at the site of the injection. Generally, the
ingredients are supplied either separately or mixed together in
unit dosage form, for example, as a dry lyophilized powder or water
free concentrate in a hermetically sealed container such as an
ampoule or sachette indicating the quantity of active agent. Where
the crystalline polymorph or amorphous form of a Compound of the
Invention is to be administered by infusion, it can be dispensed,
for example, with an infusion bottle containing sterile
pharmaceutical grade water or saline. Where the compound of the
invention is administered by injection, an ampoule of sterile water
for injection or saline can be provided so that the ingredients may
be mixed prior to administration.
[0102] It is preferred that the compositions of the invention be
administered orally. Compositions for oral delivery may be in the
form of tablets, lozenges, aqueous or oily suspensions, granules,
powders, emulsions, capsules, syrups, or elixirs, for example.
Orally administered compositions may contain one or more optionally
agents, for example, sweetening agents such as fructose, aspartame
or saccharin; flavoring agents such as peppermint, oil of
wintergreen, or cherry; coloring agents; and preserving agents, to
provide a pharmaceutically palatable preparation. Moreover, where
in tablet or pill form, the compositions may be coated to delay
disintegration and absorption in the gastrointestinal tract thereby
providing a sustained action over an extended period of time.
Selectively permeable membranes surrounding an osmotically active
driving compound are also suitable for orally administered
crystalline polymorph or amorphous form of a Compound of the
Invention. In these later platforms, fluid from the environment
surrounding the capsule is imbibed by the driving compound, which
swells to displace the agent or agent composition through an
aperture. These delivery platforms can provide an essentially zero
order delivery profile as opposed to the spiked profiles of
immediate release formulations. A time delay material such as
glycerol monostearate or glycerol stearate may also be used. Oral
compositions can include standard vehicles such as mannitol,
lactose, starch, magnesium stearate, sodium saccharine, cellulose,
magnesium carbonate, etc. Such vehicles are preferably of
pharmaceutical grade.
[0103] The amount of a crystalline polymorph or amorphous form of a
Compound of the Invention that will be effective in the treatment
of a particular disorder or condition disclosed herein will depend
on the nature of the disorder or condition, and can be determined
by standard clinical techniques. In addition, in vitro or in vivo
assays may optionally be employed to help identify optimal dosage
ranges. The precise dose to be employed in the compositions will
also depend on the route of administration, and the seriousness of
the disease or disorder, and should be decided according to the
judgment of the practitioner and each patient's circumstances.
However, suitable dosage ranges for oral administration are
generally about 0.001 milligram to 1000 milligrams of a compound of
the invention per kilogram body weight. In specific preferred
embodiments of the invention, the oral dose is 0.01 milligram to
500 milligrams per kilogram body weight, more preferably 0.1
milligram to 100 milligrams per kilogram body weight, more
preferably 0.5 milligram to 50 milligrams per kilogram body weight,
and yet more preferably I milligram to 10 milligrams per kilogram
body weight. In a most preferred embodiment, the oral dose is 1
milligram of a crystalline polymorph or amorphous form of a
Compound of the Invention per kilogram body weight. The dosage
amounts described herein refer to total amounts administered; that
is, if more than one compound of the invention is administered, the
preferred dosages correspond to the total amount of the compounds
of the invention administered. Oral compositions preferably contain
10% to 95% active ingredient by weight.
[0104] Suitable dosage ranges for intravenous (i.v.) administration
are 0.001 milligram to 1000 milligrams per kilogram body weight,
0.1 milligram to 100 milligrams per kilogram body weight, and 1
milligram to 10 milligrams per kilogram body weight. Suitable
dosage ranges for intranasal administration are generally about
0.01 pg/kg body weight to 1 mg/kg body weight. Suppositories
generally contain 0.01 milligram to 50 milligrams of a compound of
the invention per kilogram body weight and comprise active
ingredient in the range of 0.5% to 10% by weight. Recommended
dosages for intradermal, intramuscular, intraperitoneal,
subcutaneous, epidural, sublingual, intracerebral, intravaginal,
transdermal administration or administration by inhalation are in
the range of 0.001 milligram to 1000 milligrams per kilogram of
body weight. Suitable doses of the compounds of the invention for
topical administration are in the range of 0.001 milligram to 1
milligram, depending on the area to which the compound is
administered. Effective doses may be extrapolated from
dose-response curves derived from in vitro or animal model test
systems. Such animal models and systems are well known in the
art.
[0105] The invention also provides pharmaceutical packs or kits
comprising one or more containers filled with one or more
crystalline polymorph or amorphous form of a Compound of the
Invention. Optionally associated with such container(s) can be a
notice in the form prescribed by a governmental agency regulating
the manufacture, use or sale of pharmaceuticals or biological
products, which notice reflects approval by the agency of
manufacture, use or sale for human administration. In a certain
embodiment, the kit contains more than one crystalline polymorph or
amorphous form of a Compound of the Invention.
[0106] The crystalline polymorph or amorphous form of a Compound of
the Invention is preferably assayed in vitro and in vivo, for the
desired therapeutic or prophylactic activity, prior to use in
humans. For example, in vitro assays can be used to determine
whether administration of a specific compound of the invention or a
combination of compounds of the invention is preferred for lowering
fatty acid synthesis. The compounds of the invention may also be
demonstrated to be effective and safe using animal model
systems.
[0107] Other methods will be known to the skilled artisan and are
within the scope of the invention.
6.4. General Synthesis of the Compounds of the Invention
[0108] The 18-membered macrocycles and analogs thereof are produced
by fermentation. Cultivation of Dactylosporangium aurantiacum
subspecies hamdenensis AB 718C-41 NRRL 18085 for the production of
the tiacumicins is carried out in a medium containing carbon
sources, inorganic salts and other organic ingredients with one or
more absorbents under proper aeration conditions and mixing in a
sterile environment.
[0109] The microorganism to produce the active antibacterial agents
was identified as belonging to the family Actinoplanaceae, genus
Dactylosporangium (J. Antibiotics, 1987, 40: 567-574 and U.S. Pat.
No. 4,918,174). It has been designated Dactylasporangium
aurantiacum subspecies hamdenensis 718C-41. The subculture was
obtained from the ARS Patent Collection of the Northern Regional
Research Center, United States Department of Agriculture, 1815
North University Street, Peoria, Ill. 61604, U.S.A., where it was
assigned accession number NRRL 18085. The characteristics of strain
AB 718C-41 are given in the Journal of Antibiotics, 1987, 40:
567-574 and U.S. Pat. No. 4,918,174.
[0110] This invention encompasses the composition of novel
antibiotic agents, Tiacumicins, by submerged aerobic fermentation
of the microorganism Dactylosporangium aurantiacum subspecies
hamdenensis. The production method is disclosed in WO 2004/014295
A2, which is hereby incorporated by reference.
7. EXAMPLES
7.1. Preparation of the Crude Mixtures of Tiacumicins and the
Subsequent Crystallization of Certain Polymorphs of the
Mixtures
[0111] In an illustrative embodiment, a mixture of tiacumicins
containing the Compound of Formula I is prepared by a process
comprising: [0112] (i) culturing a microorganism in a nutrient
medium to accumulate the mixture in the nutrient medium; and [0113]
(ii) isolating the mixture from the nutrient medium; wherein the
nutrient medium comprises an adsorbent to adsorb the mixture.
[0114] The nutrient medium preferably comprises from about 0.5 to
about 15% of the adsorbent by weight. The absorbent is preferably
an adsorbent resin. More preferably, the adsorbent resin is
Amberlite.RTM., XAD16, XAD16HP, XAD2, XAD7HP, XAD1180, XAD1600,
IRC50, or Duolite.RTM. XAD761. The microorganism is preferably
Dactylosporangium aurantiacum subspecies hamdenensis. The nutrient
medium comprises the following combination based on weight: from
about 0.2% to about 10% of glucose, from about 0.02% to about 0.5%
of K.sub.2HPO.sub.4, from about 0.02% to about 0.5% of
MgSO.sub.4.7H.sub.2O, from about 0.01% to about 0.3% of KCl, from
about 0.1% to about 2% of CaCO.sub.3, from about 0.05% to about 2%
of casamino acid, from about 0.05% to about 2% of yeast extract,
and from about 0.5% to about 15% of XAD-16 resin. The culturing
step is preferably conducted at a temperature from about 25.degree.
C. to about 35.degree. C. and at a pH from about 6.0 to about
8.0.
[0115] Upon completion of fermentation, the solid mass (including
the adsorbent resin) is separated from the broth by sieving. The
solid mass is eluted with organic solvents such as, for example,
ethyl acetate then concentrated under reduced pressure.
7.2. Structure of R-Tiacumicin B
[0116] The structure of the R-Tiacumicin B (the major most active
component) is shown below in Formula I. The X-ray crystal structure
of the R-Tiacumicin B was obtained as a colorless,
parallelepiped-shaped crystal (0.08.times.0.14.times.0.22 mm) grown
in aqueous methanol. This x-ray structure confirms the structure
shown below. The official chemical name is
3-[[[6-Deoxy-4-O-(3,5-dichloro-2-ethyl-4,6-dihydroxybenzoyl)-2-O--
methyl-.beta.-D-mannopyranosyl]oxy]-methyl]-12(R)-[[6-deoxy-5-C-methyl-4-O-
-(2-methyl-1-oxopropyl)-.beta.-D-lyxo-hexopyranosyl]oxy]-11(S)-ethyl-8(S)--
hydroxy-18(S)-(1(R)-hydroxyethyl)-9,13,15-trimethyloxacyclooctadeca-3,5,9,-
13,15-pentaene-2-one.
##STR00009##
7.2.1 Analytical Data of R-Tiacumicin B
[0117] The analytical data of R-Tiacumicin B (which is almost
entirely (i.e., >90%) R-Tiacumicin).
[0118] mp 166-169.degree. C. (white needle from isopropanol);
[0119] [.alpha.].sub.D.sup.20-6.9 (c 2.0, MeOH);
[0120] MS m/z (ESI) 1079.7 (M+Na).sup.+;
[0121] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 7.21 (d, 1H), 6.59
(dd, 1H), 5.95 (ddd, 1H), 5.83 (br s, 1H), 5.57 (t, 1H), 5.13 (br
d, 1H), 5.09 (t, 1H), 5.02 (d, 1H), 4.71 (m, 1H), 4.71 (br s, 1H),
4.64 (br s, 1H), 4.61 (d, 1H), 4.42 (d, 1H), 4.23 (m, 1H), 4.02
(pentet, 1H), 3.92 (dd, 1H), 3.73 (m, 2H), 3.70 (d, 1H), 3.56 (s,
3H), 3.52-3.56 (m, 2H), 2.92 (m, 2H), 2.64-2.76 (m, 3H), 2.59
(heptet, 1H), 2.49 (ddd, 1H), 2.42 (ddd, 1H), 2.01 (dq, 1H), 1.81
(s, 3H), 1.76 (s, 3H), 1.65 (s, 3H), 1.35 (d, 3H), 1.29 (m, 1H),
1.20 (t, 3H), 1.19 (d, 3H), 1.17 (d, 3H), 1.16 (d, 3H), 1.14 (s,
3H), 1.12 (s, 3H), 0.87 (t, 3H);
[0122] .sup.13C NMR (100 MHz, CD.sub.3OD) .delta. 178.4, 169.7,
169.1, 154.6, 153.9, 146.2, 143.7, 141.9, 137.1, 137.0, 136.4,
134.6, 128.5, 126.9, 125.6, 124.6, 114.8, 112.8, 108.8, 102.3,
97.2, 94.3, 82.5, 78.6, 76.9, 75.9, 74.5, 73.5, 73.2, 72.8, 71.6,
70.5, 68.3, 63.9, 62.2, 42.5, 37.3, 35.4, 28.7, 28.3, 26.9, 26.4,
20.3, 19.6, 19.2, 18.7, 18.2, 17.6, 15.5, 14.6, 14.0, 11.4.
7.3. Preparation of a First Polymorph of R-Tiacumicin B
[0123] Another illustrative embodiment of the invention comprises a
process for producing a polymorph of a Compound of Formula I from a
mixture of tiacumicins comprising the steps of [0124] a) dissolving
a crude mixture of tiacumicins containing from about 76% to about
100% of a Compound of Formula I in a minimum amount of solution
comprising methanol, water, acetonitrile, acetic acid, or isopropyl
alcohol mixtures thereof; [0125] b) allowing the solution of a) to
evaporate while standing at room temperature (e.g., about
22.degree. C.) for 3 to 7 days to precipitate a first polymorph of
a Compound of Formula I; and [0126] c) separating the polymorph
from the solution by techniques known in the art.
7.3.1. Illustrative Example 1 of the Preparation of a Polymorph of
R-Tiacumicin B
[0127] After the fermentation process as described for example in
Section 7.1., the crude material was purified by reverse phase
chromatography using a Biotage Flash 75L system containing a 1.2
kg, Biotage KP-C18-HS silica column, eluted with 70:30:1,
MeOH/H.sub.2O/AcOH. The collected fractions containing 75-80% of
Compound of Formula I were combined and concentrated to one-third
of the original volume to produce a precipitate. The precipitate is
filtered and washed with water. The solid was dried under high
vacuum to afford an off-white powder. HPLC analysis showed the
powder contains about 78% of Compound of Formula I as a major
product and a mixture of tiacumicins as the minor component.
[0128] The mixture of tiacumicins containing about 78% of Compound
of Formula I (i.e., 50 mg) was dissolved in 2 mL of methanol
followed by addition of 1 mL of water. The solution was allowed to
evaporate, while standing at room temperature for 7 days to produce
a crystalline precipitate. The crystal is separated from the
solution by filtration. After methanol/water recrystallization, the
crystals contain about 90% of Compound of Formula I based on
HPLC.
7.3.2. Illustrative Example 2 of the Preparation of a Polymorph of
R-Tiacumicin
[0129] After the fermentation process as described for example in
Section 7.1., the crude material was purified by reverse phase
chromatography using a Biotage Flash 150 system containing a 3.75
kg, Biotage KP-C18-HS silica column, eluted with 52:48:1,
EtOH/H.sub.2O/AcOH. The collected fractions containing about 80-88%
of Compound of Formula I were combined and concentrated to
one-third the original volume to produce a precipitate. The
precipitate was filtered and washed with water. The solid was dried
under high vacuum. HPLC analysis showed the powder contains 85.4%
of Compound of Formula I as a major product and a mixture of
tiacumicins as the minor component.
[0130] The mixture containing about 85% of Compound of Formula I
(i.e., 1000 mg) was dissolved in 20 mL of a mixture of methanol and
water at ratios 1:1 methanol water. The solution was allowed to
evaporate/stand at room temperature for 3 days to produce a
polymorph crystalline precipitate. The crystal was separated from
the solution by filtration.
[0131] The composition obtained is a mixture containing a first
polymorph of a Compound of Formula I, and at least one of the
tiacumicin compounds based on HPLC analysis. The composition has a
melting point of 165-169.degree. C.
7.3.3. Illustrative Example 3 of the Preparation of a Polymorph of
R-Tiacumicin
[0132] After the fermentation process as described for example in
Section 7.1., the crude material was purified by reverse phase
chromatography using a Biotage Flash 75L system containing a 1.2
kg, Biotage KP-C18-HS silica column, eluted with MeOH/H.sub.2O/AcOH
67:33:4 to 70:30:1. The collected fractions containing >90% of
Compound of Formula I was combined and concentrated to one-third
volume. The precipitate was filtered and washed with water. The
solid was dried under high vacuum. HPLC analysis showed the powder
contains 94.0% of Compound of Formula I.
[0133] The solid was tested by X-ray diffraction (XRD) and
Differential Scanning calorimetry (DSC) (See FIGS. 2 and 4). The
X-ray diffraction of the solid shows peaks at angles 2.theta. of
7.7.degree., 15.0.degree., and 18.8.degree..+-.0.1 indicating the
solid is the form of a first polymorph of a Compound of Formula I.
The DSC plot shows an endothermic curve starting at about at
169.degree. C. and peak at 177.degree. C.
7.3.4. Illustrative Example 4 of the Preparation of a Polymorph of
R-Tiacumicin
[0134] After the fermentation process as described for example in
Section 7.1., the crude material was purified by reverse phase
chromatography using a Biotage Flash 75L system containing a 1.2
kg, Biotage KP-C18-HS silica column, eluted with 52:48:1,
EtOH/H.sub.2O/AcOH. The collected fractions containing >90% of
Compound of Formula I were combined, one-third volume of water was
added and left at room temperature overnight. The precipitate was
filtered and washed with water. The solid was dried under high
vacuum. HPLC analysis showed the powder contains 94.7% of Compound
of Formula I.
[0135] The powder containing 94.7% of Compound of Formula I (i.e.,
98 mg) was dissolved in 3 mL of methanol and then 1 mL of water was
added. The solution was allowed to evaporate and stand at room
temperature for 7 days to produce a crystalline precipitate. The
crystals were separated from the solution by filtration and washed
with methanol/water 3:1. The crystals were analyzed by X-ray
diffraction.
[0136] Composition of the precipitate is a mixture comprising a
Compound of Formula I based on HPLC analysis with a melting point
of 166-169.degree. C.
7.3.5. Illustrative Example 5 of the Preparation of a Polymorph of
R-Tiacumicin
[0137] After the fermentation process as described for example in
Section 7.1, the mixture was purified on a column, and a 0.06 gm of
a mixture of tiacumicins was dissolved in 16 mL of methanol and 4
mL of water in a 20 mL vial. The vial is covered with parafilm, and
pinholes were punched through. The covered vial is placed in a
desiccator and stored at room temperature for ten days. Parafilm
cover is then removed, and the vial is returned to desiccator.
Crystalline material is produced within three to five days after
the parafilm is removed. The crystalline material is washed with a
solution of methanol and water and the Compound of Formula I was
isolated in 75.6%.
[0138] X-ray powder diffraction pattern of the crystalline material
is shown in FIG. 3 included 2.theta. of 7.7.degree., 15.0.degree.,
and 18.0.degree..
7.3.6. Illustrative Example 6 of the Preparation of a Polymorph of
R-Tiacumicin
[0139] Preparation of a Polymorph From Isopropanol
[0140] After the fermentation process as described for example in
Section 7.1., the crude material was purified by reverse phase
chromatography using a Biotage Flash 150 system containing a 3.75
kg, Biotage KP-C18-HS silica column, eluted with 52:48:1,
EtOH/H.sub.2O/AcOH. The collected fractions containing 80-88% of
Compound of Formula I were combined and concentrated to one-third
of the original volume to produce a precipitate. The precipitate
was filtered and washed with water. The solid was dried under high
vacuum. HPLC analysis showed the powder contains 85.4% of Compound
of Formula I.
[0141] The powder containing 85.4% Compound of Formula I (i.e.,
2000 mg) was dissolved in 900 mL of isopropanol. The solution was
heated to increase solubility and then filtered to remove insoluble
materials. The clear solution was allowed to evaporate/stand at
room temperature for 14 days to produce a crystalline precipitate.
The crystal is separated from the solution by filtration.
[0142] Composition of the precipitate is a mixture comprising
Compound of Formula I and at least one of other related substances
based on HPLC analysis with mp of 163-165.degree. C.
[0143] X-ray diffraction of the precipitate shows peaks at angles
2.theta. of 7.6.degree. and 15.4.degree..
7.3.7. Illustrative Example 7 of the Preparation of a Polymorph of
R-Tiacumicin
[0144] After the fermentation process as described for example in
Section 7.1., and column purification, a mixture of Compound of
Formula I, >90%, 15 g) was dissolved in minimum amount of
methanol (from about 20 mL to about 30 mL), the solution was
triturated with isopropanol (.about.100 mL) to produce a polymorph.
The solid is separated from the solution by filtration with melting
point of 165-168.degree. C.
[0145] The XRD diagram shows a distinct polymorph pattern
comprising 2 theta values of 7.5.degree., 15.2.degree.,
15.7.degree., 18.6.degree. 18.7.degree..
7.3.8. Illustrative Example 5 of the Preparation of a Polymorph of
R-Tiacumicin
[0146] Preparation of a Polymorph from Acetonitrile
[0147] The mixture of tiacumicins obtained as described above and
(85.44% of Compound of Formula I, 1000 mg) was dissolved in 30 mL
of acetonitrile. The solution was allowed to evaporate and stand at
room temperature for 12 days to produce a crystalline precipitate.
The crystal is separated from the solution by filtration, and
exhibits a melting point of 165-169.degree. C.
[0148] The XRD diagram of this crystal shows the pattern of a
polymorph comprising 2 theta values of 7.8.degree., 15.1.degree.,
18.8.degree..
7.4. Preparation of Other Polymorphs of R-Tiacumicin
[0149] Another illustrative embodiment of the invention comprises a
process for producing a polymorph of a Compound of Formula I
comprising the steps of: [0150] a) dissolving crude mixture of
tiacumicins containing from about 78 to about 100% of a Compound of
Formula I in a minimum amount of ethyl acetate; [0151] b) allowing
the solution to evaporate and stand at room temperature for 3 to 7
days to precipitate a polymorph; and [0152] c) separating polymorph
from the solution
7.4.1. Illustrative Example 1 of the Preparation of a Polymorph of
R-Tiacumicin
[0153] Preparation of Polymorph from Ethyl Acetate
[0154] After the fermentation process as described for example in
Section 7.1., the crude material was purified by reverse phase
chromatography using a Biotage Flash 150 system containing a 3.75
kg, Biotage KP-C18-HS silica column, eluted with 52:48:1,
EtOH/H.sub.2O/AcOH. The collected fractions containing 70-88% of
Compound of Formula I was combined and concentrated to one-third
volume to produce a precipitate. The precipitate is filtered and
washed with water. The solid was dried under high vacuum. HPLC
analysis showed the powder contains 85.4% of Compound of Formula
I.
[0155] This crude tiacumicin mixture (1000 mg) was then dissolved
in 30 mL of ethyl acetate. The solution was allowed to evaporate
and stand at room temperature for 12 days to produce a crystalline
precipitate of Polymorph B of the Compound of Formula I. The
crystals were separated from the solution by filtration. The
crystals have a melting point of about 153-156.degree. C., which
confirm a different polymorphic form from the first polymorph.
7.4.2. Illustrative Example 2 of the Preparation of a Polymorph of
R-Tiacumicin
[0156] Preparation of a Polymorph from Methanol and
Isopropanol.
[0157] After the fermentation process as described for example in
Section 7.1, six different batches of crude material of varying
amounts of Compound of Formula I were combined such that the
combination has an average of 91% of Compound of Formula I. The
combination was dissolved in methanol and concentrated by rotary
evaporation. The concentrated solution is then mixed with
isopropanol, filtered, and dried by vacuum to produce a white
powder with a melting point of 156-160.degree. C.
[0158] X-ray powder diffraction of the white powder is shown in
FIG. 6 comprising 2 theta values of 7.5.degree., 15.4.degree., and
18.7.degree..
7.4.3. Illustrative Example 3 of the Preparation of a Polymorph of
R-Tiacumicin
[0159] Preparation of Polymorph B from Chloroform
[0160] After the fermentation process as described for example in
Section 7.1., a crude material of tiacumicins containing Compound
of Formula I was dissolved in chloroform and concentrated by
evaporation at room temperature to produce a solid with a melting
point of 156-160.degree. C.
7.4.4. Illustrative Example 4 of the Preparation of a Polymorph of
R-Tiacumicin
[0161] Preparation of a Polymorphic Form From Acetone
[0162] After the fermentation process as described for example in
Section 7.1., a crude material of tiacumicins containing Compound
of Formula I was dissolved in acetone and concentrated by
evaporation at room temperature to produce a solid with a melting
point of 156-160.degree. C.
7.5. Preparation of Amorphous Forms of Compound of Formula
[0163] Preparation of Amorphous Mixture of Tiacumicins
[0164] The amorphous mixture of tiacumicins was obtained after
column purification without any further processing steps.
Alternatively, chloroform or acetone may be added to the mixture of
tiacumicins and the solvent is evaporated to form the amorphous
product.
[0165] X-ray powder diffraction of the product exhibits no defined
diffraction peaks.
8. EXPERIMENTAL DATA
8.1. Polymorph Experimental Data
[0166] A first polymorph of a Compound of a Compound of Formula I
is characterized by Differential Scanning calorimetry ("DSC") and
powder X-Ray Diffraction ("XRD").
[0167] The DSC plot of the polymorph shows an endothermic curve at
177.degree. C.
[0168] The XRD diagram (reported in FIG. 1) shows peaks comprising
at diffraction angles 2.theta. of 7.7.degree., 15.0.degree.,
18.8.degree.. The XRD was analyzed with a Phillips powder
Diffractometer by scanning from 20 to 70 degrees two-theta at 1.0
degree per minute using Cu K-alpha radiation, at 35 kV and 20 ma.
The instrumental error (variant) is 0.04 (2 theta value).
[0169] The melting point of the mixtures containing various amounts
of Compound of Formula I is summarized in Table 1. All of the
products with at least 85% of a Compound of Formula I in the form
of a polymorph appear to have a melting point in the range of
163-169.degree. C. measured by Melting Point apparatus, MEL-TEMP
1001.
TABLE-US-00001 TABLE 1 Melting point of polymorph mixtures in
different solvent conditions Compound of Formula I Content (%) of
the crystalline Crystallization No. material Mp (.degree. C.)
Solvent 1 85 165-169 MeOH/Water 2 85 163-165 Isopropanol 3 85
164-168 Acetonitrile 4 90 165-168 MeOH/Isopropanol 5 94 166-169
MeOH/Water 6 95 166-169 MeOH/Water 7 98 163-164
MeOH/Isopropanol
[0170] Composition of the a polymorphic crystal from a mixture
comprising Compound of Formula I and optionally at least on
compound that is a mixture of tiacumicins based on HPLC analysis
with a melting point of 166-169.degree. C.
[0171] X-ray diffraction of a polymorphic crystal shows
characteristic peaks at angles 2.theta. of 7.8.degree.,
15.0.degree., 18.8.degree., and 23.9.degree.. Table 2 is a listing
of the obtained X-ray diffraction peaks for first polymorph of
R-Tiacumicin from Experiment 7.2.2.
TABLE-US-00002 TABLE 2 X-ray diffraction peaks for a First
Polymorph from Experiment 7.3.2. Two-Theta Relative Intesity 3.3568
44.0000 3.4400 47.0000 7.7815 112.0000 10.1575 32.0000 13.6023
21.0000 15.0951 139.0000 17.0178 18.0000 18.8458 36.0000 19.3771
9.0000 20.0300 16.0000 20.4842 10.0000 23.9280 136.0000 24.8338
10.0000 25.0889 19.0000 25.7256 10.0000 30.9126 75.0000 31.9970
10.0000 34.4507 30.0000
[0172] Table 3 is a listing of the obtained X-ray diffraction peaks
for Polymorph from Experiment 7.3.6.
TABLE-US-00003 TABLE 3 X-ray diffraction peaks for a Polymorph from
Experiment 7.3.6. Two-Theta Relative Intensity 3.2978 41.0000
7.5615 400.0000 9.9482 21.0000 15.4289 31.0000 22.0360 20.0000
22.5361 20.0000 24.9507 12.0000 29.5886 10.0000 34.8526 19.0000
37.7092 17.0000 40.4361 13.0000 42.2446 18.0000
8.2. Second Polymorph of R-Tiacumicin Experimental Data
[0173] A second polymorph of Compound of Formula I is also
characterized by Differential Scanning calorimetry (DSC) and powder
X-Ray Diffraction (XRD).
[0174] The DSC plot of polymorph B shows an endothermic curve at
158.degree. C. The XRD diagram (reported in FIG. 5) shows peaks
comprising at the values of the diffraction angles 2-theta of
7.6.degree., 15.4.degree. and 18.8.degree.. Polymorph B has a
melting point in the range of 153-156.degree. C. measured by
Melting Point apparatus, MEL-TEMP 1001.
[0175] It is believed that crystalline polymorphic forms of
Compounds of Formula I other than the above-discussed A and B exist
and are disclosed herein. These crystalline polymorphic forms,
including A and B, and the amorphous form or mixtures thereof
contain varying amounts of Compound of Formula I and in certain
cases mixtures of tiacumicins can be advantageously used in the
production of medicinal preparations having antibiotic
activity.
[0176] X-ray powder diffraction of the crystals is shown in FIG. 3
with peaks at angles 2.theta. of 7.5.degree., 15.7.degree., and
18.9.degree..+-.0.04 indicating the presence of Polymorph B.
[0177] The DSC plot of Polymorph B shows an endothermic curve
starting at about at 150.degree. C. and peak at 158.degree. C.
[0178] Table 4 is a summary of the various data that was isolated
for illustrative crystallization lots.
TABLE-US-00004 TABLE 4 Data Summarizing Various Lots Compound of
Formula I DSC Content Mp (.degree. C.) Crystallization No. (%)
(.degree. C.) Peak XRD (2 theta) Solvent 1 76.3 155-158 7.7, 15.0,
18.8, MeOH/Water 2 85.3 159-164 180 7.8, 14.9, 18.8, MeOH/Water 3
85.4 163-165 7.6, 15.4 Iso-propanol (IPA) 4 85.4 164-168 7.9, 15.0,
18.8 Acetonitrile 5 85.4 153-156 7.5, 15.7, 18.9 EtOAc 6 90 165-168
7.5, 15.2, 15.7, MeOH/ 18.6 Isopropanol 7 97.2 160-163 177 7.4,
15.4, 18.7 IPA 8 94.0 166-169 177 7.6, 15.1, 18.6 MeOH/Water 9 97.2
167-173 187 7.8, 14.8, 18.8 MeOH/Water 10 96.7 160 7.5, 15.4, 18.8
EtOAc 11 98.3 163-164 178 7.7, 15.0, 18.8 MeOH/IPA
[0179] The present invention is not to be limited in scope by the
specific embodiments disclosed in the examples which are intended
as illustrations of a few aspects of the invention and any
embodiments which are functionally equivalent are within the scope
of this invention. Indeed, various modifications of the invention
in addition to those shown and described herein will become
apparent to those skilled in the art and are intended to fall
within the appended claims.
[0180] A number of references have been cited, the entire
disclosures of which are incorporated herein by reference.
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