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(11) Patent Number: KE 595 (45) Date of grant: 07/08/2013
 
(12) PATENT
 
(73)    Owner:
Bayer Business Services GmbH of Law & Patents, Patents and Licensing, 51368 Leverkusen, Germany
(72) Inventors:
WINTER, Gabriele, Stieleichenstrasse 15 16567 Schonfliess, Germany; SCHNEIDER, Matthias, Wollestrasse 31, 14482 Potsdam, Germany; GOTTFRIED, Michael, Herzogstrasse 44, 42103 Wuppertal, Germany and GEISLER, Jens, Merianweg 10, 13587 Berlin, Germany
(74)    Agent/address for correspondence:
Kaplan & Stratton Advocates, P.O. Box 40111-00100, Nairobi
 
(54) Title:
N-(2-AMINOPHENYL)-44N-PYRIDINE-3-YL)-METHOXYCARBONYL-AMINOMETHYL1- BENZAMIDE (MS-275)POLYMORPH B.
(57) Abstract:
The    crstalline    Polymorph    B    of    N-(2-aminopheny1)-44N-pyridine-3-y1)methoxy-
carbonyaminomethylibenzamide (MS-275 of formula (1) is described, as well as the process for the production of said compound, and its use as a medicament for the tretment of selected diseases.
 
N-(2-aminopheny1)-4-N-(pyridine-3-y1)-methoxycarbonyl-aminomethyg-
benzamide (MS-275) polymorph B
The invention refers to the crystalline Polymorph B of N-(2-aminopheny1)-4-[1\1- (pyridine-3-yOmethoxycarbonylaminomethyl]benzamide (MS-275), the process for the production, and the use of said compound as a medicament for the treatment of selected diseases.
In EP 0 847 992 Al (which co-patent is US 6,794,392) benzamide derivatives as medicament for the treatment of malignant tumors, autoimmune diseases, de¬rmatological diseases and parasitism are described. In particular, these deriva¬tives are highly effective as anticancer drugs, preferred for the haematological malignancy and solid tumors. The preparation of N-(2-aminopheny1)-4-[N¬(pyridine-3-yl)methoxycarbonylaminomethyl]tenzamide is described on page 57, Example 48. The compound is neither purified by chromatography nor puri¬fied by treatment with charcoal. The final step of the process comprises the re-crystallization from ethanol.
Said compound has a melting point (mp) of 159 — 160 °C.
The IR spectrum shows the following bands: IR(KBr) cm-1: 3295, 1648, 1541, 1508, 1457, 1309, 1183, 742.
The data indicate the Polymorph A form.
In EP 0 974 576 B1 a method for the production of monoacylated phenylenediamine derivatives is described. The preparation of N-(2- aminopheny1)-4-[N-(pyridine-3-yl)methoxycarbonylamino-methyl] benzamide is described on pages 12 to 13, Example 6. The final step of the process com¬prises the purification of the compound via silica gel column chromatography.
Said compound has a melting point (mp) of 159 — 160 °C.
The IR spectrum shows the following bands: IR(KBr) cm-1: 3295, 1648, 1541, 1508, 1457, 1309, 1183, 742.
The data indicate the Polymorph A form.
 
2
In J. Med. Chem. 1999, 42, 3001-3003, the synthesis of new benzamide deriva-tives and the inhibition of histone deacetylase (HDAC) is described. The
process for the production of N-(2-aminophenyl)-4-[N-(pyridine-3-yl) meth-
5 oxycarbonylaminomethyl] benzamide is described. The final step of the process comprises the purification of the compound via silica gel column chromatogra¬phy (ethyl acetate).
Said compound has a melting point (mp) of 159 —160 °C.
The IR spectrum shows the following bands: IR(KBr) cm-1: 3295, 1648, 1541, 10    1508, 1457, 1309, 1183, 742.
The data indicate the Polymorph A form.
In WO 01/12193 Al a pharmaceutical formulation comprising N-(2- aminopheny1)-4-N-(pyridine-3-y1)methoxycarbonylamino-methylibenzamide is 15    described.
In WO 01/16106 a formulation comprising N-(2-aminopheny1)-44N-(pyridine-3- yl)methoxycarbonylamino-methylJbenzamide, having an increased solubility and
20 an improved oral absorption for benzamide derivatives, and pharmaceutically acceptable salts thereof are described.
In WO 2004/103369 a pharmaceutical composition is described which com¬prises histone deacetylase inhibitors. That application concerns the combined
25 use of N-(2-aminopheny1)-44N-(pyridine-3-yOmethoxycarbonylamino-methyljbenzamide together with different cancer active compounds. In fact that application is a later application, which is based on the above mentioned matter and thus concerns the Polymorph A form.
 
3
Finally, JP 2001-131130 (11-317580) describes a process for the purification of monoacylphenylenediamine derivatives. In Reference Example 2, the
process for the production of crude N-(2-aminophenyl)-4-[N-(pyridine-3-yl) meth-oxycarbonylaminomethyl] benzamide is described.
5 Said compound has a melting point (mp) of 159 — 160 °C,
The IR spectrum shows the following bands: IR(KBr) cm 1: 3295, 1648, 1541, 1508, 1457, 1309, 1183, 742.
The data indicate the Polymorph A form.
Moreover, Working Example 1 describes the purification of crude N-(2-
10 aminophenyl)-4-[N-(pyridine-3-yl) methoxycarbonylaminomethyl] benzamide in aqueous acid medium together with carbon .The final crystallization is done un¬der aqueous conditions at 40-50°C.
Following the description to that example it can be seen from the Comparative
Examples 1 — 3 that the crude N-(2-aminopheny1)-44N-(pyridine-3-y1) meth-
15 oxycarbonylaminomethyl] benzamide is not purified by dissolution under reflux conditions in either ethanol, methanol or acetonitrile followed by a recrystalliza¬tion at 2°C. As a result, these recrystallisations do not yield any pure compound.
In addition a "purification" of crude N-(2-aminophenyl)-4-[N-(pyridine-3-yl) methoxycarbonylaminomethyl] benzamide in ethanol under reflux conditions to-
20 .gether with carbon is decribed. After filtering off the carbon the compound is re¬crystallized at 2°C. The purification effect of this method is very limited. 1,1% of an impurity remain in the N-(2-aminophenyl)-4-[N-(pyridine-3-yl) methoxycar¬bonylaminomethyl] benzamide. As a result, this procedure does not yield any pure compound.
25
None of the state of the art documents refer to a polymorph B of N-(2- aminopheny1)-44N-(pyridine-3-y1)Methoxyca rbonylamino-methyl]benzamide and no physicochemical features of said compound are known.
 
4
Several biological and clinical studies have been done with N-(2-aminopheny1)- 4-[N-(pyridine-3-y1) meth-oxycarbonylaminomethyl] benzamide. For example, Kummar et al., Clin Cancer Res. 13 (18), 2007, pp 5411-5417 describe a phase I trial of N-(2-aminophenyl)-4-[N-(pyridine-3-yl) meth-oxycarbonylaminomethyl]
5 benzamide in refractory solid tumors. The compound was applied orally.
A further study in advanced refractory solid tumors or lymphoma was published by Ryan et al., J. Clin. Oncol., Vol. 23, 17, 2005, pp. 3912 — 3922 and Gore et al., Clin Cancer Res., Vol 14, 2008, pp. 4517 — 4525.
Further activity in adults with refractory and relapsed acute leukemias are pub-10    lished by Gojo et al., Blood, Vol. 109, 2007, pp. 2781-2790.
In the course of process development (scale up and modified purification proc¬ess) it has now surprisingly been found that the known form of N-(2-
aminopheny1)-4-[N-(pyridine-3-yl)methoxycarbonylamino-methyl]benzamide
15 which is described in the above mentioned state of the art, is not the thermody¬namically stable polymorph of said compound, at least not in the relevant tem-
perature range below 60°C, but is polymorph A of N-(2-aminopheny1)-4-N-(pyridine-3-yl)methoxycarbonylamino-methypenzamide.
20 This generates problems of not using the thermodynamically stable polymorph of N-(2-aminophenyI)-4-[N-(pyridine-3-yl)methoxycarbonylamino-methyl]benzamide for drug development when using polymorph A. This bears
for example the risk that the polymorph A form of N-(2-aminopheny1)-44N¬(pyridine-3-y1)methoxycarbonylamino-methyl]benzamide transforms partly or
25 completely into other polymorph forms of N-(2-aminopheny1)-44N-(pyridine-3- yl)methoxycarbonylamino-methylibenzamide (e.g. during storage as drug sub¬stance as well as drug product). However, a stable solid state form is a prereq¬uisite for developing a medicinal product because a conversion of the solid form is associated with changes in properties.
30
 
5
In addition, it was not possible to establish a reliable manufacturing process for polymorph A as pure polymorphic phase at larger scale.
An additional problem is the reliable manufacture of polymorph A with high chemical purity.
5 These problems are now solved by the thermodynamically more stable poly-morph of N-(2-aminopheny1)-44N-(pyridine-3-yOmethoxycarbonylamino-methyl]- benzamide, polymorph B, as described herein.
The inventive N-(2-aminophenyl)-4-[N-(pyridine-3-yl)methoxy-carbonylamino¬10 methyllbenzamide polymorph B of formula I
 
15 can be obtained via a process, which comprises the following process steps:
a) crude N-(2-aminopheny1)-44N-(pyridine-3- yl)methoxycarbonylaminomethyll-benzamide is suspended in water and diluted hydrochloric acid is added to the reaction mixture, at a constant in¬ternal reaction vessel temperature below 5°C, and
20    b) to said reaction mixture charcoal is added and the reaction mixture is
then stirred for -I to 20 hours at a constant temperature below 5°C, and
c) is filtered to remove the charcoal from the solution and is rinsed with wa¬ter, and
 
6
d)    while keeping the internal vessel temperature below 5°C the pH of the re¬action mixture is adjusted to > 8 with a diluted sodium hydroxide solution, and
e)    the resulting precipitated N-(2-aminopheny1)-44N-(pyridine-3- yOmethoxycarbonylaminomethyl]-benzamide is washed with water and ethanol, and is dried, and
f)    the precipitate is suspended into a mixture of ethanol and water and is heated up to a temperature of 40 - 90°C for 1 to 10 hours, and
g)    after cooling down the mixture, the resulting precipitate is rinsed with wa-
10    ter and ethanol to give the pure N-(2-aminopheny1)-44N-(pyridine-3-
y1)methoxycarbonylaminomethylkbenzamide polymorph B which is sub-sequently dried at a temperature between 30-60 °C.
The inventive process is also an object of the instant invention
15
The crude N-(2-aminopheny1)-44N-(pyridine-3-y1)methoxycarbonylaminomethyl]- benzamide of step a) can be produced according to the method described in ex¬ample 6 of EP 0974 576 B1.
20 The inventive N-(2-aminophenyl)-4-[N-(pyridine-3-yl)methoxycarbonylamino-
methyl]-tenzamide polymorph B, produced according to the instant process might have a chemically purity of at least 94%, preferred of at least 96%, more preferred of at least 98%, much more preferred of at least 99%, most preferred of at least 99.5%.
25    It should be noted that the inventive N-(2-aminopheny1)-4-N-(pyridine-3-
yl)methoxycarbonylamino-methylFbenzamide polymorph B is free of any Poly-morph A form.
 
7
For example, a higher purity can be achieved by repeating step b) and step c) of the above mentioned process if after passing step c), an insufficient purity of the N-(2-am in opheny1)-44N-(pyrid ne-3-yl)methoxycarbonyla m ino-methyli¬benzamide is determined.
5 One embodiment provides for polymorph B of N-(2-aminopheny1)-44N-(pyridine¬3-yl)methoxycarbonylamino-methyllbenzamide substantially free of any other solid state form (e.g., another polymorph).
A further embodiment provides for polymorph B of N-(2-aminopheny1)-4-1N-(pyridine-3-yl)methoxycarbonylamino-methypenzamide substantially free of
to polymorph A of N-(2-aminopheny1)-44N-(pyridine-3-yl)methoxycarbonylamino-methypenzamide.
As referred to herein, the term "substantially free" includes a solid state compo-
sition completely free of other solid state forms of N-(2-aminopheny1)-44N¬(pyridine-3-yl)methoxycarbonylamino-methyl]benzamide. The term "substan-
15 tially free" also includes solid state compositions of polymorph B that contain less than 10 %, less than 5%, less than 2%, less than 1%, or less than 0.5% of other solid state forms or other polymorphs; wherein the content of other form of of N-(2-aminopheny1)-44N-(pyridine-3-yl)methoxycarbonylamino¬methyl]benzamide is determined by various analytical methods, e.g. X-ray poW-
20 der diffraction, FT-Raman spectroscopy, IR spectroscopy, differential scanning calorimetry, microcalorimetry and solid state NMR.
In addition to polymorph B of N-(2-aminopheny1)-44N-(pyridine-3- yl)methoxycarbonyl-aminomethylibenzamide, a third anhydrous form, the poly-
25 morph C of N-(2-aminopheny1)-44N-(pyridine-3-yl)methoxycarbo-nylamino¬methyl]-benzamide and an amorphous phase were found.
The polymorph C and the amorphous phase of N-(2-aminopheny1)-44N-(pyridine-3-yl)methoxycarbo-nylamino-methyl]-benzamide are thus also an ob¬ject of the invention.
30
 
8
The polymorph B of N-(2-aminophenyI)-4-[N-(pyridine-3-yl)methoxycarbonyl-aminomethyl]benzamide is an anhydrous form and has a melting point (mp) of 156 —158 °C (see Example 2).
The polymorph C of N-(2-aminophenyl)-4-[N-(pyridine-3-yl)methoxycarbo-
5 nylamino-methyl]-benzamide is characterized by its melting point of 152 —155 °C. The amorphous phase is characterized by a glass transition tempera-
ture between 30 °C and 50 °C. Depending on heating rate the amorphous phase recrystallizes during heating into one of the polymorphs.
Beside the melting points, polymorph A, polymorph B and polymorph C are also
10 be distinguishable by the positions of the reflections in the X-ray powder diffrac¬tion (XRPD) pattern (see Example 3). The amorphous phase is characterized by the a X-ray powder diffraction pattern which shows only one ore two broad dif¬fuse maxima, but no defined XRPD reflections.
The three different polymorphs, as well as the amorphous phase, can further be 15 distinguished by their characteristic bands in the FT-Raman spectra (see Exam¬ple 4).
A differentiation between the present state of the art, polymorph A, and the new polymorph B of N-(2-aminopheny1)-44N-(pyridine-3-yOmethoxycarbonylamino-methyl]-benzamide is also possible by IR spectroscopy (see Example 5).
The polymorph B is the thermodynamically stable polymorph of N-(2- aminopheny1)-44N-(pyridine-3-yOmethoxycarbonylaminomethyl]-benzamide, at least in the relevant temperature range below 60°, which is therefore the pre¬ferred form for the use as medicament.
Thus, the crystalline N-(2-aminopheny1)-44N-(pyridine-3-yl)methoxycarbonyl¬amino-methylj-benzamide polymorph B is characterized in that its X-ray diffrac¬togram has a reflection at 2Theta = 21.1°.
 
9
Further, the crystalline N-(2-aminopheny1)-4-N-(pyridine-3-yl)methoxycarbonyl-amino-methyll-benzamide polymorph B is characterized in that its X-ray diffrac-togram has reflections at 2Theta = 21.1°, 20.4° and 27.4°.
Additional characterization is given by the Raman spectrum, wherein the crystal-
5    line N-(2-aminopheny1)-4-[N-(pyridine-3-yOmethoxycarbonylamino-methyl]-
benzamide polymorph B has its a bands at 902 cm-1, 3036 cm-1, 1639 cm-1 and 916 cm-1.
The IR-spectrum of polymorph B of N-(2-aminopheny1)-44N-(pyridine-3-yl-methoxycarbonylamino-methyll-benzamide shows the following characteristic
10        bands: IR (KBr) cm-I: : 3349, 3311, 1705, 1641, 1262 and 751. and IR (AIR)
cm 1: 3349, 3309, 1702, 1638, 1260 and 749.
Thus, the characteristics of the pure crystalline N-(2-aminopheny1)-4-[1■1- (pyridine-3-yl)methoxycarbonylamino-methyl]-benzamide polymorph B are its X-ray diffractogram which has a reflection at 2Theta = 21.1°, 20.4° and 27.4°, and
15 its FT-Raman spectrum which has bands at 1705 cm-1, 1641 cm-1, 1262 cm-land 751 cmI, and its IR (ATR) spectrum with bands at 1702 cm-1, 1638 cm-I, 1260 cm' and 749 cm-1
Due to the fact that the polymorph B is the thermodynamically stable polymorph of MS-275, at least in the relevant temperature range below 60°, its manufacture
20 in highly pure form at larger scale is easier than the manufacture of polymorph A of MS-275.
The N-(2-aminopheny1)-4-[N-(pyridine-3-yl)methoxycarbonylaminomethyl]- benzamide polymorph B can be used for the production of a medicament for the 25 treatment of different diseases, as well as in established test systems.
For example, N-(2-aminopheny1)-441-(pyridine-3-yOmethoxycarbonylamino-methyl]benzamide polymorph B can be used as medicament for the treatment of malignant tumors, auto — immune diseases, dermatological diseases and para-
30    SitiSM.
 
10
The term "malignant tumors" comprises hematologic malignancy such as acute leukaemia, malignant lymphoma, multiple myeloma and macroglobulinemia as well as solid tumors such as colon cancer, cerebral tumor, head and neck tu¬mor, breast carcinoma, pulmonary cancer, oesophageal cancer, gastric cancer,
5    hepatic cancer, gallbladder cancer, bile duct cancer, pancreatic cancer, nesi-
dioblastoma, renal cell carcinoma, adrenocortical cancer, urinary bladder carci-
noma, prostatic cancer, testicular tumor, ovarian carcinoma, uterine cancer, chorionic carcinoma, thyroid cancer, malignant carcinoid tumor, skin cancer, ma¬lignant melanoma, osteogenic sarcoma, soft tissue sarcoma, neuroblastoma,
10 Wilms tumor and retinoblastoma.
The term "autoimmune diseases" comprises rheumatism, diabetes, systemic lu-
pus erythematodes, human autoimmune lymphocytic lymphadenopathy, im-munoblastic lymphadenopathy, Crohn disease and ulcerative colitis.
The term "dermatologic diseases" comprises psoriasis, acne, eczema and atopic 15    dermatitis.
The term "parasitism" includes diseases such as malaria caused through vermi¬nation.
The invention thus further comprises the use of N-(2-aminopheny1)-44N1-
20 (pyridine-3-yl)methoxycarbonylamino-methyl]-benzamide polymorph B for the treatment of said diseases, as well as the use of of N-(2-aminopheny1)-4411- (pyridine-3-yl)methoxycarbonylamino-methylFbenzamide polymorph B for the production of a medicament for the treatment of said diseases.
25 The use of N-(2-aminopheny1)-44N-(pyridine-3-yOmethoxycarbonylamino-methyli-benzamide polymorph B as medicament for the treatment of malignant tumors is preferred.
 
11
The manufacture of the medicaments/formulations may be performed according to methods known in the art. Commonly known and used adjuvants, as well as further suitable carriers or diluents may be used.
5 Suitable carriers and adjuvants may be such as recommended for pharmacy, cosmetics and related fields in: Ullmann's Encyclopedia of Technical Chemistry, Vol. 4, (1953), pp. 1-39; Journal of Pharmaceutical Sciences, Vol. 52 (1963), p. 918ff; H.v.Czetsch-Lindenwald, "Hilfsstoffe far Pharmazie und angrenzende Ge¬biete"; Pharm. Ind. 2, 1961, p.72ff; Dr. H.P. Fiedler, Lexikon der Hilfsstoffe fur
10 Pharmazie, Kosmetik und angrenzende Gebiete, Cantor KG, Aulendorf in Wurt-temberg, 1971, which are hereby incorporated by reference.
Said medicaments and formulations are also an object of the instant invention.
15 For a therapeutical effect the sensible dose is different and depends on the con-centration in the pharmaceutical composition, the host, the form of application and the intensity of the disease which is treated.
The invention also comprises pharmaceutical compositions, which can be pre-
20    pared by known methods of preparing galenics for oral, enteral, parenteral, e.g.
intravenous, intraperitoneal, intramuscular, subcutaneous or percutaneous ap-plication. The inventive combination can also be implanted into tissue.
Thus, the invention comprises the use of the N-(2-aminopheny1)-4411-(pyridine¬3-yOmethoxycarbonylamino-methyl}-benzamide polymorph B composition for
25 enteral administration, such as nasal, buccal, rectal or, expecially, oral admini¬stration, and for parenteral administration, such as intravenous, intramuscular or subcutaneaous administration, to warm-blooded animals, especially, humans, are especially preferred. The compositions comprise the N-(2-aminophenyI)-4- [N-(pyridine-3-yl)methoxycarbonylamino-methyl]-benzamide polymorph B alone
30 or, preferably, together with a pharmaceutically acceptable diluent and/ or car¬rier. The dosage of the active ingredient depends upon the disease to be treated
 
12
and upon the species, gender, age, weight, and individual condition, the individ¬ual pharmacokinetic data, and the mode of administration.
Preferred amounts of N-(2-aminophenyl)-4-[N-(pyridine-3-yl)methoxycarbonyl-
5 aminomethyll-benzamide polymorph B are in the range of 0.0001 to 100 mg/kg per day, preferred 0.001 to 10 mg/kg per day, more preferred 0.01 to 1 mg/kg per day and most preferred 0.05 to 0.5 mg/kg per day.
Depending on the amount, necessary for the treatment of a patient it might be useful to apply a defined amount of active compound on one or on several days 10    in the same or different dose.
N-(2-aminopheny1)-441-(pyridine-3-yOmethoxycarbonyl-aminomethyll-benzamide polymorph B can also be used prophylactically or especially thera-
peutically, to a process for the preparation of a composition (especially in the form of compositions for the treatment of tumors) and to a method of treating of
is a number of diseases, especially tumor diseases, more especially those men¬tioned hereinabove.
In the preferred embodiment, the N-(2-aminopheny1)-44N-(pyridine-3- yOmethoxycarbonyl-aminomethylFbenzamide polymorph B is suitable for ad-ministration to a warm-blooded animal, especially humans or commercially use-
20 ful mammals suffering form a disease especially a neoplastic disease, and com¬prises an effective quantity of said compound for the treatment of the disease, together with at least one pharmaceutically acceptable carrier.
The N-(2-aminopheny1)-44N-(pyridine-3-yl)methoxycarbonyl-arninomethylF
25 benzamide polymorph B can also be administered in the form of tablets, film coated tablets, wafers, suppositories, pills, dragees, gel capsules, granules, sup¬positories, implants, injectable sterile aqueous or oily solutions, suspensions or
emulsions, ointments, creams, gels, patches for transdermal administration, formulations suitable for administration by inhalation, for instance nasal sprays.
30
As combination with at least one pharmaceutically acceptable carrier, said com-
bination comprises the N-(2-aminophenyl)-4-[N-(pyridine-3-yl)methoxycarbonyl-
 
13
aminomethyl]-benzamide polymorph B from approximately 0.1% to approxi¬mately 95%, single-dose administration forms comprising in the preferred em¬bodiment from approximately 1% to approximately 20% active ingredient. Unit
dose forms are, for example, coated and uncoated tablets, ampoules, vials,
5 suppositories or capsules. Further dosage forms are, for example, ointments, creams, pastes, foams, tinctures, lip-sticks, drops, sprays, dispersions, etc. Ex¬amples are capsules containing from about 0.05 mg to about 1.0 g active ingre¬dients.
10 For the preparation of the pharmaceutical compositions for oral administration, the active agents suitable for the purposes of the present invention as defined above can be admixed with commonly known and used adjuvants and carriers such as for example, gum arabic, talcum, starch, sugars like e.g. mannitose, methyl cellulose, lactose, gelatin, surface-active agents, magnesium stearate,
is aqueous or non-aqueous excipients, paraffin derivatives, crosslinking agents,
dispersants, emulsifiers, lubricants, conserving agents and flavoring agents (e.g., ethereal oils).
In the pharmaceutical composition, the N-(2-aminopheny1)-44N-(pyridine-3- yl)methoxycarbonyl-aminomethyll-benzamide polymorph B may be dispersed in 20    a microparticle, e.g. a nanoparticulate, composition.
The N-(2-aminopheny1)-44N-(pyridine-3-y1)methoxycarbonylaminomethyli-benzamide polymorph B can thus be combined with one or more of those physiologically acceptable pharmaceutical adjuvants and carriers to give an ac-
25 ceptable formulation as medicament.
Further pharmacologically effective adjuvants and carriers are, for example, de-
scribed in Remington's Pharmaceutical Science, 15th ed. Mack Publishing Company, Easton Pennsylvania (1980), which is hereby incorporated by refer¬ence
30
In order to further enhance the bioavailability of the N-(2-aminopheny1)-4-[N-(pyridine-3-yOmethoxycarbonylaminornethyl]-benzamide polymorph B, the corn-
 
14
pound suitable for the purposes of the present invention as defined above can also be formulated as cyclodextrin clathrates by reacting them with a-, 11- or y¬cyclodextrines or derivatives thereof according to the method as disclosed in WO 96/02277.
5
For parenteral administration the N-(2-aminopheny1)-4-N-(pyridine-3- yOmethoxycarbonylamino-methylJ-benzamide polymorph B suitable for the pur¬poses of the present invention as defined above can be dissolved or suspended in a physiologically acceptable diluent, such as e.g., oils with or without solubi-
10 lizers, surface-active agents, dispersants or emulsifiers. As oils for example and without limitation, olive oil, peanut oil, cottonseed oil, soybean oil, castor oil and sesame oil may be used.
The N-(2-aminopheny1)-4-EN-(pyridine-3-y1)methoxycarbonylaminomethylF
15 benzamide polymorph B according to the present invention can also be adminis¬tered via a depot injection or an implant preparation, optionally for sustained de-livery of the active agent(s).
Implants can comprise as inert materials e.g. biologically degradable polymers 20 or synthetic silicones such as e.g. silicone rubber.
For percutaneous applications, the N-(2-aminopheny1)-441•1-(pyridine-3- yl)methoxycarbonylamino-methylj-benzamide polymorph B may also be formu¬lated into adhesives.
25
The preferred mode of administration is oral administration.
Formulations comprising the N-(2-aminopheny1)-44N-(pyridine-3- yOmethoxycarbonylaminomethyli-benzamide polymorph B can be prepared in a
30 manner known per se, for example by means of conventional mixing, granulat¬ing, coating, dissolving or lyophilizing processes.
 
15
Preference is given to the use of solutions of the N-(2-aminopheny1)-4-N¬(pyridine-3-Amethoxycarbonylaminomethyll-benzarnide polymorph B, and also suspensions or dispersions, especially isotonic aqueous solutions, dispersions or suspensions which, for example in the case of lyophilized compositions com-
5 prising the active ingredients alone or together with a carrier, for example manni¬tol, can be made up before use. The pharmaceutical compositions may be steril¬ized and/or may comprise excipients, for example preservatives, stabilizers, wet¬ting agents and/or emulsifiers, solubilizers, salts for regulating osmotic pressure and/or buffers and are prepared in a manner known per se, for example by
to    means of conventional dissolving and lyophilizing processes. The said solutions
or suspensions may comprise visosity-increasing agents, typically sodium car-boxymethylcellulose, carboxymethylcellulose, dextran, polyvinylpyrrolidone, or gelatins, or also solubilizers, for example Twee„ 80 [polyoxyethyl¬ene(20)sorbitan mono-oleate; trademark (RTM) of ICI Americas, Inc. USA].
15
Suspensions in oil comprise as the oil component the vegetable, synthetic, or semi-synthetic oils customary for injection purposes. In respect of such, special mention may be made of liquid fatty acid esters that contain as the acid compo-nent a long-chained fatty acid having from 8 to 22, expecially from 12 to 22, car-
20 bon atoms, for example lauric acid, tripdecylic acid, myristic acid, pentadecylic acid, palmitic acid, margaric acid, stearic acid, arachidic acid, behenic acid or corresponding unsaturated acids, for example oleaic acid, elaidic acid, erucic a¬cid, brassidic acid or linoleic acid, if desired with the addition of anti-oxidants, for example vitamine E, a-carotene or 3,5-di-tert-butyl-4-hydroxytoluene. The alco-
25 hol component of these fatty acid esters has a maximum of 6 carbon atoms and is a monovalent or polyvalent, for example a mono-, di- or trivalent, alcohol, for example methanol, ethanol, propanol, butanol or pentanol or the isomers there¬of, but especially glycol and glycerol. As fatty acid esters, therefore, the following are mentioned: ethyl oleate, isopropyl myristate, isopropyl palmitate, „Labrafil M
30 2375"RTM (polyoxyethylene glycerol trioleate from Gattefosse, Paris), „Labrafil M 1944 CS"RTM (unsaturated polyglycolized glycerides prepared by alcoholysis of apricot kernel oil and consisting of glycerides and polyethylene glycol ester; Gat¬tefosse, France), „Labrasol"RTM (saturated polyglycolized glycerides prepared by
 
16
alcoholysis of TCM and consistitn of glycerides and polyethylene glycol ester; Gattefosse, France), and/or „Miglyol 812"RTM (triglyceride of saturated fatty acids of chain length C8 to C12 from HCJIS AG, Germany), but especially vegetable oils such as cottonseed oil, almond oil, olive oil, castor oil, sesame oil, soybean oil
5 and more expecially groundnut oil.
The manufacture of injectable preparations is usually carried out under sterile conditions, as is filling, for example into ampoules or vials, and the sealing of the containers.
10
Pharmaceutical compositions for oral administration can be obtained, for exam¬ple, by combining the N-(2-aminopheny1)-44N-(pyridine-3-yl)methoxycarbonyi-aminomethyl}-benzamide polymorph B with one or more solid carriers, if desired granulating a resulting mixture, and processing the mixture of granules, if de-
15        sired or necessary, by the inclusion of additional excipients, to form tablets or
tablet cores.
Suitable carriers are especially fillers, such as sugars, for example lactose, sac-charose, mannitol or sorbitol, cellulose preparations, and/or calcium phosphates,
20 for example tricalcium phosphate or calcium hydrogen phosphate, and also bin¬ders, such as starches, for example corn, wheat, rice or potato starch, methyl-cellulose, hydroxypropyl methylcellulose, sodium carboxymethyl-cellulose, and/or polyvinylpyrrolidone, and/or, if desired, disintegrators, such as the above-
mentioned starches, also carboxymethyl starch, crosslinked polyvinylpyrroli-
25 done, alginic acid or a salt thereof, such as sodium alginate. Additional excipi¬ents are especially flow conditioners and lubricants, for example silicic acid, talc, stearic acid or salts thereof, such as magnesium or calcium stearate, and/or po¬lyethylene glycol, or derivatives thereof.
30 Tablet cores can be provided with suitable, optionally enteric, coatings through the use of, inter elle, concentrated sugar solutions, which may comprise gum arabic, talc, polyvinylpyrrolidone, polyethylene glycol and/or titanium dioxide, or coating solutions in suitable organic solvents or solvent mixture, or for the prepa-
 
17
ration of enteric coatings, solutions of suitable cellulose preparations, such as acetyl cellulose phthalate or hydroxypropylmethylcellulose phthalate. Dyes or pigments may be added to the tablets or tablet coatings, for example for identifi-
cation purposes or to indicate different doses of the N-(2-aminophenyl)-4-[N-
s    polymorph B.
Pharmaceutical compositions for oral administration also include hard capsules
consisting of gelatin, and also soft, sealed capsules consisting of gelatin and plasticizer, such as glycerol or sorbitol. The hard capsules may contain the N-(2-
10 aminopheny1)-4-1N-(pyridine-3-yl)methoxycarbonylaminomethyli-benzamide polymorph B in the form of granules, for example in admixture with fillers, such
as corn starch, binders, and/or glidants, such as talc or magnesium stearate, and optionally stabilizers. In soft capsules, the N-(2-aminopheny1)-4-1N-(pyridine¬3-yl)methoxycarbonylaminomethyll-benzamide polymorph B is preferably dis-
cs        solved or suspended in suitable liquid excipients, such as fatty oils, paraffin oil or
liquid polyethylene glycols or fatty acid esters of ethylene or propylene glycol, to
which stabilizers and detergents, for example of the polyoxyethylene sorbitan fatty acid ester type, may also be added.
20 Other oral dosage forms are, for example syrups prepared in customary manner which comprise the active ingredient, for example, in suspended form and in a concentration of about 5% to 20%, preferably about 10%, or in similar concen-tration that provides a suitable single dose, for example, when administered in measures of 5 or 10 ml. Also suitable are, for example, powdered or liquid con-
25 centrates for the preparation of shakes, for example in milk. Such concentrates may also be packaged in single-dose units.
Pharmaceutical compositions suitable for rectal administration are, for example, suppositories that consist of a combination of the N-(2-aminophenyl)-4-[N-
30 (pyridine-3-Amethoxycarbonylaminomethy1]-benzamide polymorph B and a suppository base. Suitable suppository bases are, for example, naturral or syn-thetic triglycerides, paraffin hydrocarbons, polyethylene glycols or higher alka-nols.
 
18
For parenteral administration, aqueous solutions of the N-(2-aminopheny1)-44N-(pyridine-3-yl)methoxycarbonylaminomethylltenzamide polymorph B in water-soluble form, for example of a water-soluble salt, or aqueous injection suspen-
5 sions that contain viscosity-increasing substances, for example sodium car¬boxymethylcellulose, sorbitol and/or dextran, and, if desired, stabilizers, are es¬pecially suitable. The N-(2-aminopheny1)-44N-(pyridine-3-yOmethoxycarbonyl-aminomethyl]-benzamide polymorph B, optionally together with excipients, can also be in the form of a lyophilizate and can be made into a solution before par-
10 enteral administration by the addition of suitable solvents.
Solutions such as are used, for example, for parenteral administration can also be employed as infusion solutions.
15 Preferred preservatives are, for example, antioxidants, such as ascorbic acid, or micro-bicides, such as sorbic acid or benzoic acid.
The present invention relates especially also to the use of the combination, as such or in the form of a pharmaceutical formulation with at least one pharmaceu-
20 tically acceptable carrier for the therapeutic and also prophylactic management of one or more of the diseases mentioned hereinabove, especially a neoplastic disease.
Optionally, the N-(2-aminopheny1)-44N-(pyridine-3-ypmethoxycarbonylamino¬25 methyl]-benzamide polymorph B can be combined with one or more pharmaco¬logically active agents.
For example, the compounds of this invention can be combined with known anti-hyper-proliferative, antiinflammatory, analgesic, immunoregulatory, diuretic, an-
30        tiarrhytmic, anti-hypercholsterolemia, anti-dyslipidemia, anti-diabetic or antiviral
agents, and the like, as well as with admixtures and combinations thereof.
 
19
A combination of the N-(2-aminopheny1)-44N-(pyridine-3-yOmethoxycarbonyl-amino-methyl]-benzamide polymorph B together with cytotoxic agents is pre-ferred.
5 The additional pharmaceutical agent can be aldesleukin, alendronic acid, alfaf-
erone, alitretinoin, allopurinol, aloprim, aloxi, altretamine, aminoglutethimide, amifostine, amrubicin, amsacrine, anastrozole, anzmet, aranesp, arglabin, arse¬nic trioxide, aromasin (exemestan), 5-azacytidine, azathioprine, BCG or twice
BCG, bestatin, betamethasone acetate, betamethasone sodium phosphate,
10    bexarotene, bleomycin sulfate, broxuridine, bortezomib, busulfan, calcitonin,
campath, capecitabine, carboplatin, casodex, cefesone, celmoleukin, cerubidine, chlorambucil, cisplatin, cladribine, 2-cladribine, clodronic acid, cyclophos¬phamide, cytarabine, dacarbazine, dactinomycin, DaunoXome, decadron, decadron phosphate, delestrogen, denileukin diftitox, depo-medrol, deslorelin,
15        dexrazoxane, diethylstilbestrol, diflucan, docetaxel, doxifluridine, doxorubicin,
dronabinol, DW-166HC, eligard, elitek, ellence, emend, epirubicin, epoetin alfa,
epogen, eptaplatin, ergamisol, erlotinib (Tarceva), estrace, estradiol, estra-mustine phosphate sodium, ethinyl estradiol, ethyol, etidronic acid, etopophos, etoposide, fadrozole, farston, filgrastim, finasteride, fligrastim, floxuridine, flu-
20 conazole, fludarabine, 5-fluorodeoxyuridine monophosphate, 5-fluorouracil (5-FU), fluoxymesterone, flutamide, formestane, fosteabine, fotemustine, ful¬vestrant, gammagard, gemcitabine, gemtuzumab, gleevec, gliadel, goserelin, granisetron HCI, histrelin, hycamtin, hydrocortone, eyrthro-hydroxynonyladenine, hydroxyurea, ibritumomab tiuxetan, idarubicin, ifosfamide, interferon alpha, inter-
25        feron-alpha 2, interferon alfa-2A, interferon alfa-2B, interferon alfa-n1, interferon
alfa-n3, interferon beta, interferon gamma-la, interleukin-2, intron A, iressa,  iri-
notecan, kytril, lentinan sulphate, letrozole, leucovorin, leuprolide, leuprolide acetate, levamisole, levofolinic acid calcium salt, Ievothroid, levoxyl, lomustine,
lonidamine, marinol, mechlorethamine, mecobalamin, medroxyprogesterone
30 acetate, megestrol acetate, melphalan, menest, 6-mercaptopurine, Mesna, methotrexate, metvix, miltefosine, minocycline, mitomycin C, mitotane, mitoxan-trone, Mod renal, Myocet, nedaplatin, neulasta, neumega, neupogen, nilutamide,
nolvadex, NSC-631570, OCT-43, octreotide, ondansetron HCI, orapred, ox-
 
20
aliplatin, paclitaxel, pediapred, pegaspargase, Pegasys, pentostatin, picibanil, pilocarpine HCI, pirarubicin, plicamycin, porfimer sodium, prednimustine, predni-solone, prednisone, premarin, procarbazine, procrit, raltitrexed, rebif, rhenium-186 etidronate, rituximab, roferon-A, romurtide, salagen, sandostatin, sargramo-
5    stim, semustine, sizofiran, sobuzoxane, solu-medrol, sparfosic acid, stem-cell
therapy, streptozocin, strontium-89 chloride, synthroid, tamoxifen, tamsulosin, tasonermin, tastolactone, taxotere, teceleukin, temozolomide, teniposide, testos-
terone propionate, testred, thioguanine, thiotepa, thyrotropin, tiludronic acid, topotecan, toremifene, tositumomab, trastuzumab, treosulfan, tretinoin, trexall,
10 trimethylmelamine, trimetrexate, triptorelin acetate, triptorelin pamoate, UFT, uridine, valrubicin, vesnarinone, vinblastine, vincristine, vindesine, vinorelbine, virulizin, zinecard, zinostatin stimalamer, zofran, A131-007, acolbifene, actim¬mune, affinitak, aminopterin, arzoxifene, asoprisnil, atamestane, atrasentan, BAY 43-9006 (sorafenib), avastin, CCI-779, CDC-501, celebrex, cetuximab,
15 crisnatol, cyproterone acetate, decitabine, DN-101, doxorubicin-MTC, dSLIM, dutasteride, edotecarin, eflornithine, exatecan, fenretinide, histamine dihydro-chloride, histrelin hydrogel implant, holmium-166 DOTMP, ibandronic acid, inter-feron gamma, intron-PEG, ixabepilone, keyhole limpet hemocyanin, L-651582, lanreotide, lasofoxifene, libra, lonafarnib, miproxifene, minodronate, MS-209, Ii-
20 posomal MTP-PE, MX-6, nafarelin, nemorubicin, neovastat, nolatrexed, oblimersen, onco-TCS, osidem, paclitaxel polyglutamate, pamidronate disodium, PN-401, QS-21, quazepam, R-1549, raloxifene, ranpirnase, 13-cis -retinoic acid, satraplatin, seocalcitol, T-138067, tarceva, taxoprexin, thymosin alpha 1, tiazo¬furine, tipifarnib, tirapazamine, TLK-286, toremifene, TransMID-107R, valspodar,
25    vapreotide, vatalanib, verteporfin, vinflunine, Z-100, zoledronic acid or combina-
tions thereof.
The additional pharmaceutical agent can also be gemcitabine, paclitaxel, cis-
platin, carboplatin, sodium butyrate, 5-FU, doxirubicin, tamoxifen, etoposide,
30 trastumazab, gefitinib, intron A, rapamycin, 17-AAG, U0126, insulin, an insulin
derivative, a PPAR ligand, a sulfonylurea drug, an a-glucosidase inhibitor, a biguanide, a PTP-1B inhibitor, a DPP-IV inhibitor, a 11-beta-HSD inhibitor, GLP-
 
21
1, a GLP-1 derivative, GIP, a GIP derivative, PACAP, a PACAP derivative, se-cretin or a secretin derivative.
Optional anti-hyper-proliferative agents which can be added to the composition
5 include but are not limited to compounds listed on the cancer chemotherapy drug regimens in the 11th Edition of the Merck Index, (1996), which is hereby in-
corporated by reference, such as asparaginase, bleomycin, carboplatin, car-mustine, chlorambucil, cisplatin, colaspase, cyclophosphamide, cytarabine, dacarbazine, dactinomycin, daunorubicin, doxorubicin (adriamycine), epirubicin,
10 etoposide, 5-fluorouracil, hexamethylmelamine, hydroxyurea, ifosfamide, iri-
notecan, leucovorin, lomustine, mechlorethamine, 6-mercaptopurine, mesna, methotrexate, mitomycin C, mitoxantrone, prednisolone, prednisone, procarba¬zine, raloxifen, streptozocin, tamoxifen, thioguanine, topotecan, vinblastine, yin¬cristine, and vindesine.
15
Other anti-hyper-proliferative agents suitable for use with the composition of the invention include but are not limited to those compounds acknowledged to be
used in the treatment of neoplastic diseases in Goodman and Gilman's The Pharmacological Basis of Therapeutics (Ninth Edition), editor Molinoff et al.,
20 publ. by McGraw-Hill, pages 1225-1287, (1996), which is hereby incorporated by reference, such as aminoglutethimide, L-asparaginase, azathioprine, 5-
azacytidine cladribine, busulfan, diethylstilbestrol, 2',2'-difluorodeoxycytidine, docetaxel, erythrohydroxynonyl adenine, ethinyl estradiol, 5-fluorodeoxyuridine, 5-fluorodeoxyuridine monophosphate, fludarabine phosphate, fluoxymesterone,
25 flutamide, hydroxyprogesterone caproate, idarubicin, interferon, medroxypro-gesterone acetate, megestrol acetate, melphalan, mitotane, paclitaxel, pen-tostatin, N-phosphonoacetyl-L-aspartate (PALA), plicamycin, semustine, teni-poside, testosterone propionate, thiotepa, trimethylmelamine, uridine, and vi-norelbine.
30
Other anti-hyper-proliferative agents suitable for use with the composition of the invention include but are not limited to other anti-cancer agents such as epothilone and its derivatives, irinotecan, raloxifen and topotecan.
 
22
It should be noted that some of the above mentioned compounds are trade-marks (RTM)
5 Generally, the use of cytotoxic and/or cytostatic agents in combination with the N-(2-aminopheny1)-4411-(pyridine-3-y1)methoxycarbonyl-amino-methyll¬benzamide polymorph B of the present invention will serve to:
I) yield better efficacy in reducing the growth of a tumor or even eliminate the to    tumor as compared to administration of either agent alone,
ii) provide for the administration of lesser amounts of the administered chemo-therapeutic agents,
15 iii) provide for a chemotherapeutic treatment that is well tolerated in the patient with fewer deleterious pharmacological complications than observed with
single agent chemotherapies and certain other combined therapies,
iv)    provide for treating a broader spectrum of different cancer types in mam¬20    mats, especially humans,
v)    provide for a higher response rate among treated patients,
vi)    provide for a longer survival time among treated patients compared to stan-25    dard chemotherapy treatments,
vii)    provide a longer time for tumor progression, and/or
30    yield efficacy and tolerability results at least as good as those of the
agents used alone, compared to known instances where other cancer agent combinations produce antagonistic effects.
 
23
The instant invention further comprises the combination of the N-(2-amino¬phenyl)-44N-(pyridine-3-ypmethoxycarbonylamino-methyl]-benzamide poly-morph B together wit one ore more regiments of cytotoxic agents, for example, Folfox4 (fluorouracil/ leucovorin/ oxaliplatin); Folfiri (leukovorin, 5-fluorouracil, iri-
s notecan); DHAP (cisplatin/ cytarabine/ dexamethasone), CEOP (cyclophos¬phamide/ eprirubicin/ vincristine/ prednisone), CHOP (cyclophosphamid, doxorubicin, vincristine and prednisone), FLAG (fludarabine/ cytarabine/ filgras-
tim), M & P (mitoxantrone + prednisone; or melphalan + prednisone), ABMC (doxorubicin/ carmustine/ melphalan/ cyclophosphamide), ICE (ifosfamide, car-
10 boplatin and etoposide), DVP (daunorubicin, vincristine and prednisone), ATRA (all-trans retinoic acid), ABVD (bleomycin, dacarbazin, doxorubicin and vincris-
tine), COP (cydophosphamide, vincristine and prednisone), VAD (vincristine, adriamycin and dexametasone) and MOPP (mechlorethamine, prednisone, pro-cabazine and vincristine).
15
The cytotoxic agents itself or from the regimens are either commercially avail-, able or can be prepared by standard literature procedures.
The N-(2-aminopheny1)-4-[N-(pyridine-3-y1)methoxycarbonylaminomethyl]-
20 benzamide polymorph B can be applied in an pharmaceutically effective amount together with one or more of these cytotoxic agents simultaneously, separately or sequentially.
The amounts (a "pharmaceutically effective amount") of the combined active a-
25 gents to be administered vary within a broad range and depend on the condition to be treated and the mode of administration. They can cover any amount effi¬cient for the intended treatment. Determining a "pharmaceutically effective a¬mount" of the combined active agent is within the purview of a person skilled in the art.
30
Those combinations which comprise the N-(2-aminopheny1)-4-[N-(pyridine-3- ypmethoxy-carbonylaminomethyl]-benzamide polymorph B together with cyto-toxic and/ or cytostatic agents are also matter of the instant invention.
 
24
Examples
Example 1
Process for the production of N-(2-aminopheny1)-4-IN-(pyridine-3-y1)- 5 methoxycarbonylaminomethyll-benzamide, polymorph B form.
30 kg of crude N-(2-aminopheny1)-44N-(pyridine-3- ypmethoxycarbonylaminomethyl]-benzamide (MS-275, crude) was transferred
into a reaction vessel and approximately 300 kg of water have been added
10 (suspended). To that reaction mixture approximately 18 kg of hydrochloric acid, 36% (w/w), diluted in approximately 66 kg of water, have been added, while keeping the internal temperature constantly below 5°C. Under control of the in-
ternal temperature, 15 kg charcoal was added and the reaction mixture was stirred for approximately 10 hours.
15 After that reaction time the reaction solution was filtered to remove the charcoal from the solution and was accordingly rinsed with approximately 90 kg of water.
The purity of N-(2-aminopheny1)-44N-(pyridine-3-yl)methoxycarbonylamino¬methylj-benzamide was determined. If the purity was insufficient, again 15 kg charcoal was added to the filtrate and stirred for further 10 hours, and again the
20 reaction solution was filtered to remove the charcoal from the solution and was accordingly rinsed with approximately 90 kg of water.
While keeping the internal temperature below 5°C the pH of the reaction mixture was adjust to 8 by using a diluted sodium hydroxide solution. After said treat¬ment the N-(2-aminopheny1)-4-[N-(pyridine-3-yOmethoxycarbonylaminomethyl]-
25 benzamide product, which has been precipitated, was washed with approxi¬mately 120 kg water and with approximately 68 kg ethanol.
The precipitate was then dried at a temperature between 30-60 °C. After the precipitate was dried, it was suspended in a mixture of 5-fold (e.g. approximately 63 kg) ethanol and of 7-fold (e.g. approximately 111 kg) water, and was heated
30 up to 40-90°C for 5 hours. After that reaction time the reaction mixture was
 
25
cooled down. The precipitate was rinsed with approximately 120 kg of water and with approximately 52 kg of ethanol.
The N-(2-aminopheny1)-4-N-(pyridine-3-yl)methoxycarbonylaminomethyli-benzamide product was dried at a temperature between 30-60 °C.
5
Yield: 9 kg — 24 kg (30 % - 80% of theory) Melting Point:: 156 — 158 °C
Example 2
to Analysis of the N-(2-aminophemil)-4-1N-(Pyridine-3-yl)methoxycarbonyl¬aminomethyll-benzamide polymorphs and of the amorphous phase by dif¬ferential scanning calorimetry (DSC)
The DSC traces were recorded with a heating rate of 5 K/min in aluminium pans under nitrogen atmosphere. The determined melting temperatures of the three 15    polymorphs are given in the following Table 1.
Table 1
Polymorph A    Polymorph B    Polymorph C
       
160 °C ± 1 K    157 °C ± 1 K    152 °C — 155 °C

20
According to a person ordinary skilled in the art, the determined melting tem-perature depends on the experimental conditions, especially on the used heat¬ing rate. In addition, the melting temperature is influenced by the chemical pu¬rity of the material. The reported melting temperature were determined on
25 batches with a purity of at least 98.5 %.
 
26
Example 3
Analysis of the N-(2-aminoPherlY1)-44N-(PYridine-3-y1)methoxycarbonyl¬aminomethyll-benzamide polymorphs and of the amorphous phase by X-ray powder diffraction
5
The X-ray powder diffraction data were recorded at room temperature using ger¬manium-monochromatized CuKa1-radiation = 1.5406 A). The 2Theta scans were performed using the small linear position sensitive detector with an angular resolution of 0.08° between 2Theta 5 35° (stepwidth 0.5°), at room tempera-
lo    tore.
The 2 Theta values of the strongest reflections of the three polymorphs in the X-ray powder pattern are given in the following Table 2.
Table 2
15
Polymorph A    Polymorph B    Polymorph C
       
18.4    9.2    17.7
18.8    18.1    18.4
19.1    19.4    18.8
20..9    .    20.0    19.2.
22.6    20.4    20.0
26.4    21.1    22.0
26.7    22.1    22.3
27.2    25.8    23.2
    27.4    23.4

One of ordinary skill in the art will appreciate that an X-ray diffraction pattern 20 may be obtained with a measurement error that is dependent upon the meas-
 
27
urement conditions. In particular, it is generally known that intensities in an X-ray diffraction pattern may fluctuate depending upon crystal habitus of the material and measurement conditions employed. Additionally, a measurement error of diffraction angle Theta for a conventional X-ray diffraction pattern at a given
5 temperature is typically about ± 0.1 0, and such degree of measurement error
should be taken into account as pertaining to the diffraction angles. Conse-quently, any crystal form that provides X-ray diffraction patterns that is substan-tially identical as to those disclosed in the accompanying Figures falls within the scope of the present invention.
10
 
28
Example 4
Analysis of the N-(2-aminophenyl)-4-IN-(pyridine-3-y1)methoxycarbonyl-aminomethyll-benzamide polymorphs and the amorphous phase by FT-Raman spectroscopy
5
The FT-Raman spectra were recorded using a resolution of 2 and 64 sans at a laser power of 250 mW
The results of the Raman analysis are shown in Table 3. The data show the wave numbers (cm-1) of characteristic bands in the FT-Raman spectra of Poly-to morph A, Polymorph B and Polymorph C and the amorphous phase.
Table 3
Wave number of characteristic FT-Raman bands (cm '1)

Polymorph A    Polymorph B    Polymorph C    Amorphous
            Phase
           
           
3061    3075    3076    3062
3048    3063    3050    1649
1613    1639    1629    1613
1262    1613    1613    1598
1041    1328    1329    1322.
1034    1299    1311    1256
936    1293    1297    1041
908    1040    1040    907
893    1033    908    776
776    916    783   
    902    775   
    778       
 
29
According to a person ordinary skilled in the art, the acceptable tolerance for the wave number shift is ± 2 cm-1 dependent on the used equipment and measure¬ments conditions. Consequently, any solid state form that provides a FT-Raman spectra that is substantially identical as to those disclosed in the accompanying
s    Figures falls within the scope of the present invention.
Example 5
Analysis of the N-(2-aminopheny1)-441•14pyridine-3-yl)methoxycarbonyl-10 aminomethyll-benzamide polymorph B by IR spectroscopy
The IR spectra of the polymorph B were recorded using diffuse reflection (KBr) and ATR.. The major infrared bands and their assignments are summarized in the following Table 4.
Table 4
Wave number of characteristic IR bands of polymorph B (cm    )
       
    KBr        ATR
3349,    3311,    3216    3349, 3309
    1705        1702
    1641        1638
    1262        1260
    751        749

15
According to a person ordinary skilled in the art, the acceptable tolerance for the wave number shift is ± 2 cm-1 dependent on the used equipment and measure¬ments conditions. Consequently, any solid state form that provides a FT-Raman spectra that is substantially identical as to those disclosed in the accompanying
20    Figures falls within the scope of the present invention.
 
30
Example 6
Biological activity of the N-(2-aminopheny1)-44N-(pyridine-3 yl)methoxy-
carbonylaminomethyll-benzamide polymorph B in the DU-145 human  prostate carcinoma model in nude mice
5
Nude mice were orally treated after tumor transplantation with 10, 20 or 30 mg/kg of N-(2-aminophenyl)-4-[N-(pyridine-3 yl)methoxy-carbonylaminomethyl]- benzamide polymorph B. The active compound was applied in 30% HPRCD. As control, a vehicle of DMSO/ ethanol 1:1 in 0,085% Myrj/NaCl/ 30% HPI1CD was
10 used. The tumor growth was determined after 27 to 56 days after tumor trans¬plantation.
The results of that experiment are shown in Fig. 8.
According to the results, one can see that there is a clear inhibition with regard to the tumor growth if N-(2-aminophenyl)-4-[N-(pyridine-3 yl)methoxy-carbonyl¬15 aminomethyl]-benzamide polymorph B is applied.
 
31
Description of the figures
Fig. 1 shows a DSC trace of Polymorph A and Polymorph B of N-(2- aminopheny1)-44N-(pyridine-3-ypmethoxycarbonylaminomethylibenzamide (MS-275).
5 Fig. 2 shows a DSC trace of Polymorph C of N-(2-aminopheny1)-44N-(pyridine-3-yOmethoxycarbonylaminomethylpenzamide (MS-275).
Fig. 3 shows a DSC trace of the amorphous phase of N-(2-aminopheny1)-44N¬(pyridine-3-y1)methoxycarbonylaminomethyl]benzamide (MS-275).
Fig. 4 shows the X-ray powder diffraction pattern of Polymorph A of N-(2-
10 aminophenyI)-4-[N-(pyridine-3-yl)methoxycarbonylaminomethyl]benzamide (MS-275).
Fig. 5 shows the X-ray powder diffraction pattern of Polymorph B of N-(2- aminopheny1)-44N-(pyridine-3-11)methoxycarbonylaminomethylibenzamide (MS-275).
15 Fig. 6 shows the X-ray powder diffraction pattern of Polymorph C of N-(2- aminophenyt)-44N-(Pyridine-34)methoxycarbonylaminomethyl]benzamide (MS-275).
Fig. 7 shows the X-ray powder diffraction pattern of the amorphous phase of N¬(2-aminopheny1)-44N-(pyridine-3-Amethoxycarbonylaminomethyl]benzamide 20 (MS-275).
Fig. 8 shows the FT-Raman spectrum of Polymorph A of N-(2-aminopheny1)-4- [N-(pyridine-3-yl)methoxycarbonylaminomethyl]benzamide (MS-275).
Fig. 9 shows the FT-Raman spectrum of Polymorph B of N-(2-aminopheny1)-4- [N-(pyridine-3-ypmethoxycarbonylaminomethyl]benzamide (MS-275).
25 Fig. 10 shows the FT-Raman spectrum of Polymorph C of N-(2-aminopheny1)-4- [N-(pyridine-3-yl)methoxycarbonylaminomethyl]benzamide (MS-275).
 
32
Fig. 11 shows the FT-Raman spectrum of the amorphous phase of N-(2- aminopheny1)-44N-(pyridine-3-ypmethoxycarbonylaminomethyl]benzamide (MS-275).
Fig. 12 shows the IR spectrum (KBr) of Polymorph B of N-(2-aminophenyl)-4-[N¬s (pyridine-3-yl)methoxycarbonylaminomethyl]benzamide (MS-275), and the major infrared bands and their assignments.
Fig. 13 shows the IR spectrum (ATR) of Polymorph B of N-(2-aminopheny1)-4- [N-(pyridine-3-yl)methoxycarbonylaminomethyl]benzamide (MS-275), and the major infrared bands and their assignments.
10    Fig. 14 shows the biological activity of the inventive Polymorph B of N-(2-
aminopheny1)-44N-(pyrid ine-3-yl)methoxycarbonylami nomethylibenzamide (MS-275) (ZK 244894), applied in different concentrations against a control (Vehicle¬DMSO).
15
 
35
5. Crystalline N-(2-a inophenyl)-4-[N-(pyridine-311)methoxycarbonylamino-methyl]benzamide polymorph B according to claim 1, characterized in that its X-ray diffractogram has a reflection at 2Theta 21,1'.
s 6. Crystalline N-(2-aminophenyl)-44N-(pyridine-3-yl)methoxycarbonylmino¬methyli-benzamide polymorph B according to claim 1, characterized in that its X-ray diffractogram has reflections at 2Theta 21.1°, 20.4' and 27.4°.
7.    Crystalline N-(2-aminophenyI)-4-[N-(pyridine-3-yl)methoxycarbonylamino¬io    methylj-benzamide polymorph B according to claim 1, characterized in that
the Raman spectrum has a band at 902 cm-1.
8.    Crystalline N-(2-aminophenyl)-44N-(pyridine-3-Amethoxycarbonylamino-methy1]-benzamide polymorph B according to claims 1, characterized in that
15        the Raman spectrum has bands at 902 cm-1, 34§ cm-I, 1639 cm-1 and 916
crn1
9.    Crystalline N-(2-aminopheny1)-4414-(pyridine-3-yl)methoxycarbonylamino-methylFbenzamide polymorph B according to claim 1, characterized in that
20        the IR spectrum (ATR) has bands at 1702 cm-1, 3309 cm-1, 1638 cm-1,
1260 cm-1 and 749 cm-I.
10.    Crystalline N-(2-aminopheny1)-44N-(pyridine-3-yl)methoxycarbonylamino-methyll-benzamide polymorph B according to claim 1, characterized in that
25        the IR spectrum (KBr) has bands at 1705 cm-1, 3311 cm-1, 1641 cm-1,
1262 cm-1 and 751 cm-1.
 
36
11.Crystalline N-(2-aminopheny1)-44N-(pyridine-3-yl)methoxycarbonylamino-methyq-benzamide polymorph B according to claim 1, characterized in that its X-ray diffractogram has a reflection at 2Theta 21,1', 20,4 and 27.4' and the Raman spectrum has bands at 902 cm-1, 30.g'cm-1, 1639 cm-1 and
s        916 cm-1, and the IR (AIR) spectrum has bands at 1702 cm-1, 3311 cm-1,
1641 cm-1, 1262 cm-1 and 751 cm-1.
12.    Use of N-(2-aminophenyl)-4-N-(pyridine-311)methoxycarbonylamino-methyl]benzamide polymorph B for the production of a medicament for the
lo        treatment of malignant tumors, auto — immune diseases, dermatological dis-
eases and parasitism.
13.    A composition comprising the N-(2-aminopheny1)-44N-(pyridine-3- y1)methoxy-carbonylamino-methyllbenzamide polymorph B alone or in corn-15    bination with a pharmaceutically acceptable diluent and/ or carrier.
14.    A combination comprising the N-(2-aminopheny1)-44N-(pyridine-3- y1)methoxy-carbonylaminomethyll-benzamide polymorph B together with one or more cytotoxic or cytolytic agents.
20
15.A combination according to claim 13, characterized in that the N-(2- aminopheny1)-4-114-(pyridine-3-y1)methoxy-carbonylaminomethylj-benzamide polymorph B is for simultaneous, separate or sequential administration to¬gether with the cytotoxic or cytolytic agents.
 
Fig. 'I
 

 
I
 
Polymorph A-
 

 
ES    90    OS    100    10S 110 in 120    Lib
 
Fig. 2
endo
5
mw    Polymorph C   
        160    165    °C
85
12    90    95    100    105    110    115    120    125    130    135    140    145    150    155   
    riiiiiiiiiiiiiiiiiiiiiiiiIiiiiiiriiiiiiilitiilitiiiiiiiliiiiiiiiiiiiiiitiiiiisiiiriis
13    14    15    16    17    18    19    20    21    22    23    24    25    26    27    28    min

 

 

 
Fig. 5
 

                Polymorph B
    100.0.                                   
    80.0.                                   
    60.0.                                   
    40.0                                        
    20.0.                                           
3.0                   
    0.0    
7.0    15.0    *    19.0    '23.0    27.0    2Th eta*
 
Fig. 6
 
Polymorph C
 
100.0
 

 
20.0
 
0.0    •    •    •    •
3.0    7.0    11.0    15.0    19.0    23.0    .27.0'    31.0 2Theta
 

 
Fig. 8
 
 
Fig. 9
 
Polymorph B
0.!
0
1"..
 
C■1
O
 
\k,
 
Afi
 
4
 
) V
 
A,.
 

 
1    I    I    1    1
3000    2500    2000    1500    1000
Wavenurnber cm-1
 
Fig. 10
P (7) c • a)
F3
e               
    o -
,g1
— -
q _
,. _
0    Polymorph C    0
\ AN                \itkiV                i
.,    N.       
                                ryW                   
        3500
3000    2500    2000    1500    1000    500
Wavenumber cm-1   
 
Fig. 11
-4-
0
oi
.--
C
O
6
P
(75
i 8 C
1 ec
8
c5
8
c;
0,
0
ci
0
O
0
3500               
    Amorphous    phase    .                        Mil           
    I
3000    I
2500    I    1
2000    1500    1000    500
Wavanurnber cm-1   
 

 
Fig. 13
 

100,0, 95 90 SS SO 75 70 65
60
55
50
16T
45                                                                           
                                                                           
                                                                           
                                                                            
                                                                           
                                                                           
                                                                           
                                                                           
        3349    3309                                                           
                                                                           
                                                                           
                                                                           
                                                                           
                                                                           
                                                                           
                                                                           
                                                                           
                                                                           
                                                                           
                                                                           
                                                                           
40 35 30 25 20 05 DO 5                                                                           
                                                                           
                            1038                                       
                                                                           
                                                                           
                                                            1250               
                    1702                                                   
                                                        749       
                                                                           
0,0
4030:0    3600    3200    26120    2400    2000    1SC0 •    1600    8400    1200    1043    SOO    650,0
-cakI
 
Fig. 14
 
DU-145 human prostate carcinoma model in nude mouse
 
27    32    35    39    42    46    53    56
 
-DNISO/Et0H 1:1 in 0,085% Myrj/NaCU30% HPBCD
—x—ZK 244894 30m g/kg in 30% HPBCD po daily
—11—ZK 244894 20m glkg in 30% HPBCD po daily
 
days [after tumor transplantation]
—o—ZK 244894 10mg/kg in 30% HPBCD po daily

 

 

 

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