FORM 2
THE PATENT ACT 1970 (39 of 1970)
&
The Patents Rules, 2003
COMPLETE SPECIFICATION
(See section 10 and rule 13)
1. TITLE OF THE INVENTION:
"Synergetic formulation for Anti-tuberculosis"
2. APPLICANT (S)
(a) NAME: Genesen Labs ltd.
(b) NATIONALITY: An Indian Company incorporated under the Indian Companies ACT 1956
(c) ADDRESS: Genesen Labs ltd.,
R-75,TTC Industrial estate, Thane-Belapur road, Navi Mumbai India 400701. Tel No. 91-22-66888720. Fax No. 91-22-66888730.
3. PREAMBLE TO THE DESCRIPTION
The following specification particularly describes the invention and the manner in which it is to be performed.

Synergetic formulation for Anti-tuberculosis
Field of Invention
The present invention relates in general to the field of tuberculosis treatment and more particularly, provides an anti-tuberculosis composition comprising an antitubercular, bacteriostatic compound such as thiacetazone and antitubercular, bactericidal compound such as isoniazide, having a synergistic inexpensive and additive anti-tuberculosis effect. The present invention further provides, a pharmaceutical oral composition for use in anti-tuberculosis in mammals, including humans. Furthermore the present invention provides the process for manufacturing composition,
Background of Invention
Tuberculosis, caused by the bacterium mycobacterium tuberculosis, is an important communicable disease worldwide. Mycobacterium is a genus of bacteria which has special cell membrane structures different from other bacteria. This renders most antibiotics unable to enter the bacterial cells, leading to the failure of antibiotics to inhibit the growth of the bacteria. Tuberculosis, therefore, requires special drugs or combination of drugs for treatment.

Anti-tuberculosis drugs can be divided into two groups. The first line drugs are highly effective and of relatively low toxicity. The second line drugs are less effective and/or of relatively high toxicity, and are used when the bacteria resist the first line drugs. There are five first line drugs, which are isom'azid, rifampin, pyrazinarnide, ethambutol and streptomycin. Standard tuberculosis treatment requires four of these drugs which normally include isom'azid and rifampin with two other drugs, usually pyrazinarnide and ethambutol or streptomycin. The six month treatment starts with these four drugs for two months, followed by treatment with isoniazid and rifampin for four months.
This is because only isom'azid and rifampin are highly effective in killing the bacteria. When M. tuberculosis shows resistance to either pyrazinarnide, ethambutol or streptomycin, the treatment requires the switch to the second line drugs but the patient may still be able to complete the treatment within six months. On the other hand, if the organisms are resistant to isoniazid or rifampin, even the switch to other effective drugs may not render successful treatment within six months. The treatment may need to be lengthened up to eighteen months especially if the organisms resist rifampin so, there is need to explore the effective combination to resist the tuberculosis.

M. tuberculosis is considered a multi-drug resistant strain when it resists both isoniazid and rifampin. Multi-drug resistant tuberculosis is a very serious public health problem because it cannot be cured during the normal course of treatment or perhaps not at all. This is due to the fact that the bacteria have developed resistance to other drugs during the treatment. The patients may have no serious symptoms even though the treatment cannot eliminate the bacteria because the drugs may have control over the organisms to some extent.
The patients can therefore transmit the resistant strains to other people.The limited number of highly effective drugs is a major problem in tuberculosis control. Although isoniazid and rifampin were discovered over 30 years ago, resistant strains have since emerged. There is thus a need to identify new and effective drugs against Mycobacterium tuberculosis
A serious concern in antitubercular therapy is the emergence of multi-drug resistant (MDR) strains, and more recently, extensively drug-resistant (XDR) strains of Mycobacterium tuberculosis (M. tb). Strains of M. tb resistant to isoniazid and rifampicin, important components in the first-line of drug treatment, are categorized as MDR, while XDR strains of M. tb. are defined as those that are also resistant to at least three of the six classes of second-line

drugs (aminoglycosides, polypeptides, fluoroquinolones, thioamides, cycloserine and p-aminosalicylic acid), which seriously limits treatment options. According to a recent WHO report, 10% of MDR cases were XDR, across all geographical regions surveyed, thus posing the threat of an untreatable global epidemic.
Therefore, the need for rapid and continued progress in understanding the mechanism of action of the inexpensive antitubercular agents and the discovery of new cellular drug targets remain ever present. Combined dosage forms with fixed doses of anti-TB drugs, which include products of different groups, or the patient is given a combination of both anti-TB drugs of different classes.
Many Researchers have suggested several different approaches to enhance the effectiveness of treatment outcomes, including different methods of immunomodulation and immunotherapy.
In Chinese patent publication no. CN 1097321, suggested a Chinese medicinal composition for the treatment of tuberculosis wherein the method emphasis on the composition for pulmonary tuberculosis is prepared from Gecko, Pseudobulbos Blatille (Rhizoma Blatille ) extract Radix stemonea extract, carpex et plastron testudinus, carapex trionycis, cornu cervi pantroticnum

etc.... But the major drawback of this therapy is not cost effective due to the commercial unavailability of the above said extract.
The Russian patent No. RU 22253460, teaches the method for the treatment of pulmonary tuberculosis, including chemotherapy combinations of TB drugs such as Isoniazid and Rifampicin, characterized in that additionally administered zinc sulfate at a daily dose 230-35 mg/kg two - three-time ingestion rate of 21-28 days, thereby reducing the duration of chemotherapy due to pronounced anti-apoptotic effect on T-lymphocytes.
The Russian patent No. RU 2182483 proposed a combined anti-TB drugs, isoniazid and contains pyrazinamide, ethambutol hydrochloride. However, the presences in its composition of only two active components are not completely removing the above disadvantages of combination therapy; it requires the effective combination therapy.
In 1954 Kushner et al, J. Am. Chem. Soc. 77:1152-1155, reported the use of ethyl mercaptan and related compounds in experimental treatment of tuberculosis. Isopropyl thiopyrazinoate applied subcutaneously exhibited activity in a standardized mouse test. However, the authors attributed this activity to the release of ethyl mercaptan, not to the pyrazinoyl residue. Brown

et al, J. Am. Chem. Soc. 76:3860 (1954) also reported that ethyl mercapto compounds had antituberculosis activity, thus supporting the Kushner et al. assertion that the activity of ethyl thiolpyrazinoate was due to ethyl mercaptan and not the pyrazinoyl residue. The only suggestion that pyrazinoic acid esters might have some value in tuberculosis therapy is found in Solomons and Spoerri, J. Am. Chem, Soc. 75:679 (1953). In the course of evaluating esters of pyrazinoic and pyrazine-2,3-dicarboxylic acids as local anaesthetics, the authors learned of the effectiveness of pyrazinamide as a tuberculostatic agent. The authors tested their anaesthetic compounds for in vitro activity against Mycobacterium tuberculosis H37RV and reported that a few were active, including N,N-dimethyl-2-aminoethyl pyrazinoate. No further work appears to have been done with this compound, however. In addition, effectiveness against other mycobacteria, including pyrazinamide-resistant M. tuberculosis, would not have been obvious on the basis of this isolated in vitro test.
In U.S. Pat No. US5104875, described to reduce the development of drug resistance of Mycobacterium tuberculosis have been proposed multi-TB drugs that contain three or more active ingredients.

In Russian Patent No. RU 195937, 2003 proposed a combined anti-TB drug containing as combination of isoniazid, rifampin, pyrazinamide, ethambutol and pyridoxine, as well as an excipient metotsel - methyl ether of cellulose containing 14-30% methoxyl groups. Further, Studies have found that the purpose of preparations containing zinc, patients with pulmonary tuberculosis leads to rapid negativatsii phlegm and improves the radiographic readings (Karuadi E, Mest CT, Schultnick M, et al / A double blipd, rlasebo-soptgolled studu ° F vitamip A apd zips surrlemeptatiop regsops ip with ip tubersulosis Ipdopesia: effests on slipisal gesropse apd putritiopal status. Am J Slip Nutr.2002; 75:720-727).
In formulations of combined composition of three or more active ingredient exert the mutual influence of active ingredient significantly reduces the bioavailability of the active ingredient, which reduces the effectiveness of TB control and is the reason for the return of the disease and the development of secondary drug resistance.

Thus, an important aspect in the selection of the active ingredients of the components of combined anti-TB drugs is an account of their interaction and compatibility in the process of formulation and storage.
Summery Of Invention
The present invention relates to the synergetic combination of anti-tuberclusis drugs comprising a therapeutically effective amount of thiacetazone and isoniazide or a pharmaceutically acceptable non- toxic salt thereof.
Further, according to the present invention there is providing the process for preparation of an anti-tuberclusis pharmaceutical composition comprising the combination of Thiacetazone and Isoniazide and by using a dry granulation method, a wet granulation method or a direct compression method with an optionally the film coating on tablet core.
In addition to this, the present invention further extend to the blend of finely powdered active ingredient about to 100 to 900 urn by normal grinding and optionally pulverized or micronised to achieve the particular particle size which concomitantly having the emphasis of bioavailability in VITRO and in VIVO studies.

Detail Description of Invention
Unless defined otherwise, technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art in which this invention belongs. Any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention.
Unless stated to the contrary, any use of the words such as "including," "containing," "comprising," "having" and the like, means "including without limitation" and shall not be construed to limit any general statement that it follows to the specific or similar items or matters immediately following it. Embodiments of the invention are not mutually exclusive, but may be implemented in various combinations.
The described embodiments of the invention and the disclosed examples are
given for the purpose of illustration rather than limitation of the invention as
set forth the appended claims.
For purposes of the present invention, the following terms are defined below. The terms "administering to", "administration of and "administering a" compound, combination or composition should be understood to mean

providing a compound, combination or composition or a prodrug of a compound
to the patient in need of treatment.
The term "excipient (s)" means a component of a pharmaceutical product or
composition that is not the active ingredient, such as a filler, diluents, carrier,
and so on. The excipient(s) that are useful in preparing a pharmaceutical
composition are preferably pharmaceutically acceptable.
The term "pharmaceutically acceptable" as used herein means that the carrier, filler, diluents, binders, disintegrants and lubricants or excipient must be compatible with the other ingredients of the formulation and not deleterious to the recipients thereof.
The term "pharmaceutical composition" is intended to encompass a product comprising the active ingredient(s), pharmaceutically acceptable excipient(s) as well as any product which results, directly or indirectly, from combination, complexation or aggregation of any two or more of the ingredients, or from dissociation of one or more of the ingredients and/or excipinte(s), or from other types of reactions or interactions of one or more of the ingredients and/or excipient(s). Accordingly, the pharmaceutical compositions of the present invention encompass any composition made by admixing the active ingredient, additional active ingredient(s), and pharmaceutically acceptable e

xcipients.
The term "synergy" or "synergistic" includes but is not limited to (i) a quantity or amount, whereby a particular compound, active ingredient or drug can be administered in a lesser dosage when present in combination with one or more other active ingredients, compounds or drugs than the dosage
required when it is administered
individually; and/or (ii) nature/time, whereby a particular active ingredient, compound, or drug present in a combination either exhibits improved efficacy or results in reduced time of treatment than when it is administered individually.
The present invention provides the formulation of synergetic anti-tuberculosis drugs such as effective combination of line 1 and line 3 drugs witch effective against bacterium Mycobacterium tuberculosis in form of oral formulation such as tablet or film coated tablet.
In preferred embodiment according to the invention, the combination of first line and third line anti-tuberculosis therapy has great emphasis in terms of synergetic action and therapy cost in tuberculosis, because the therapy required several doses for several months to get the complete cure for patients. The first line drugs for anti-tuberculosis are isoniazide, rifampicin,

pyrazinamide and ethambutol in a single dose or fixed dose combination but normally in single dose therapy the time required six to nine months.
n another way the combination or fixed dose combination with third line drugs would be effective against the tuberculosis wherein the third line drugs are rifabutine, clarithromycine, linezolid, thiacetazone, thioridazine, arginine and vitamine D. But in our observation it is found that the combination or fixed dose combination of thiacetazone and isoniazide is most effective combination about to the synergetic action as well as cost effective therapy due to the duration of therapy and inexpensive active materials.
Thiaceatzone chemically known as
N[4[[{Aminothioxomethyl)hydrazono]methyl]phenyl]acetamide; which is in form of pale yellow crystals with bitter taste and this crystal is soluble in hot alcohol; very sparingly sol in cold alcohol. Insoluble in water, acetone, benzene, carbon tetrachloride, chloroform, carbon disulfide, petroleum ether. Practically insoluble in the other common organic solvents except glycols.
Isoniazide chemically known as isonicotinohydrazide colorless or white crystals or white crystalline powder. It is odorless and slowly affected by exposure to air and light. It is freely soluble in water, sparingly soluble in alcohol, and slightly soluble in chloroform and in ether.

The composition of thiacetazone and isoniazide comprises the in a weight ratio range is about to 1:4 to 4:1 respectively but preferably 1:2 to 2:1 respectively.
This invention also provides the method of treatment or control of tuberculosis in mammals, comprising the therapeutically effective amount of composition.
As one alterative composition:
a) A thiacetazone or pharmaceutically acceptable salt thereof, is present in amount of 50 mg, 75 mg, 150 mg and 300 mg;
b) A isoniazide or pharmaceutically acceptable salt thereof, is present in amount of 100 mg, 150 mg, 300 mg and 600 mg;
In preferred embodiment, the method of this invention comprises the synergetic formulation for composition of line 1 and line 3 anti-tuberculosis drug such as thiaceatzone and isoniazide by accompanying the pharmaceutically acceptable excipient(s) used in pharmaceutical formulation e.g. for oral administration.
In further preferred embodiment according to the synergetic formulation may be in the form of tablet or film coated tablet, the formulation comprising The tablet formulation according to the present invention wherein tablet core comprising therapeutically equivalent thiaceatzone and isoniazide in the finely

powder particles with particles size 100 to 900 µm but preferably 100 to 200 urn and maximum residue on a 40 mesh sieve of up to 5% and further
excipient(s) that are suitable for production of granules and lubrication which further to compression to form tablet core.
The suitable method for preparation of film coated tablet of thiaceatzone and isoniazide within in the scope of description of invention comprising the steps of dry mixing with active ingredients, granulation, lubrication and compression to form tablet core which concomitantly to further film coating wherein, the suitable pharmaceutically accepted excipiente(s) for the production of granules. The active ingredients i.e. thiacetazone and isoniazide are finely powdered or optionally pulverized or micronized about to particle size 100 to 900 urn but preferably 100 to 200 um blend which further dry mixed with filler and disintegrents such as lactose, dextrose, saccharose, glucose, sorbitol, mannitol, xylitol, potato starch, maize starch, rice starch, wheat starch or, crosscarmalose sodium, microcrystalline cellulose but preferably the combination with maize starch and microcrystalline cellulose in 1 to 15 % weight of tablet core.

Granulation may be by using binders such as gelatin, tragacanth, agar, alginic acid, cellulose ethers, for example methylcellulose, carboxymethylcellulose or hydroxypropylmethylcellulose,
polyethylene glycols or ethylene oxide homopolymers, especially having a degree of polymerisation of approximately from 2.0x10 3 to 1.0x10 5 and an approximate molecular weight of about from 1.0x10 5 to 5.0x10 6' for example excipient(s) known by the name Polyox® (Union Carbide), polyvinylpyrrolidone (PVP k-30) or povidones, especially having a mean molecular weight of approximately 1000 and a degree of polymerization of approximately from 500 to 2500, and also agar or gelatin; but preferably the polyvinylpyrrolidone (PVP k-30) alone or mixture thereof in 0.1 to 10% weight of tablet core.
The lubrication and compacting the granules may be those which are suitable for the conventional direct compression method for examples by dry lubricants such as potato, wheat and maize starch, microcrystalline cellulose, HPMC for example commercial products available under the trademarks Avicel®, Filtrak®, Heweten® or Pharmacel®, highly dispersed silicon dioxide, for example Aerosil®, mannitol, lactose, and also polyethylene glycol, especially having a molecular weight of from 4000 to 6000, crosslinked polyvinylpyrrolidone (Polyplasdone® XL or Kollidon® CL), crosslinked carboxymethylcellulose (Acdisol® CMC-XL), carboxymethylcellulose [Nymcel®,

for example ZSB-10, (Nyma)], hydroxypropylmethylcellulose, for example the quality HPMC 603, carboxymethyl starch [Explotab® (Mendell) or Primojel®
(Scholtens)], microcrystalline cellulose, for example Avicel® PH 102, dicalcium phosphate, for example Emcompresse or talcum powder. The addition of small amounts of, for example, lubricants, such as magnesium stearate, is also advantageous. But preferably the combination pack consists of talcum powder, in 0.1 to 5% weight of tablet core, sodium starch glycolate in 10 to 60% weight of tablet core and magnesium stearate in1 to 10 % weight of tablet core for easy compression and less de-dusting.
Furthermore, the above preferred pharmaceutically acceptable excipiente(s) are suitable for the production of film coated {thiacetazone and isom'azide) tablet with taking in to account of the stability and compatibility with active ingredients. Granules may be produced in a manner known per se, for example using wet granulation methods known for the production of "built-up" granules or "broken-down" granules.
Methods for the formation of built-up granules may operate continuously and comprise, for example simultaneously spraying the granulation mass with granulation solution and drying, for example in a drum granulator, in pan granulators, on disc granulators, in a fluidized bed, by spray-drying or spray-

solidifying, or operate discontinuously, for example in a fluidized bed, in a batch mixer or in a spray-drying drum.
Preferred are methods for the production of broken-down granules, which may be carried out discontinuously and in which the granulation mass first forms a wet aggregate with the granulation solution, which aggregate is then comminuted or formed into granules of the desired particle size and the granules then being dried. Suitable equipment for the granulation step is planetary mixers, low and high shear mixers, wet granulation equipment including extruders and spheronisers include.
Compression to form tablet cores may be carried out in conventional tableting machines or rotary tableting machine. The tablet cores may be of various shapes, for example round, oval, oblong, cylindrical etc., and various sizes, depending on the amount of thiacetazone and isoniazide.
The film coatings on tablet core by Water- permeable or organic solvent-permeable film-forming materials are, for example, hydrophilic mixtures of polyvinylpyrrolidone or of a copolymer of polyvinylpyrrolidone and polyvinyl acetate with hydroxypropylmethylcellulose, polyethylene glycol or mixtures of shellac with hydroxypropylmethylcellulose, polyvinyl acetate or copolymers

thereof with polyvinylpyrrolidone, or mixtures of water-soluble cellulose derivatives, such as hydroxypropylmethylcellulose, and water-insoluble ethylcellulose.
Elastic, film-like materials are especially hydrophilic, partially etherified cellulose derivatives. Hydrophilic, partially etherified cellulose derivatives are, for example, lower alkyl ethers of cellulose having an average degree of molar substitution (MS) that is higher than one and lower than three and an average degree of polymerization of approximately from 100 to 5000 and pigments for coating such as iron oxide and titanium dioxide or a mixture thereof .
The exact dose of thiacetazone and isoniazide and the particular formulation to be administered depend upon a number of factors, e.g. the condition to be treated, the desired duration of treatment and the rate of release of the thiacetazone and isoniazide. For example, the amount of thiacetazone and isoniazide required and the release rate thereof may be determined by in vitro or in vivo techniques, determining how long a particular active agent concentration in the blood plasma remains at an acceptable level for a therapeutic effect.

The following examples illustrate the invention.
Example 1
Tablet core Mg Mg Mg Mg
Thiaceatzone 50 75 150 300
Isoniazide 100 150 300 600
MCCP 40 40 50 55
Maize starch 60 56.8 60 65
PVPK-30 5.2 5.2 10 15
Talcum powder 5.2 5.3 16 20
Aerosil 1.2 1.5 2 3
Sodium starch 30 32 35 40
glycolate
Magnesium 2 3 3.5 5
stearate
Total (Tablet core 266.6 340.00 595.00 1067.00
wt.)
Film coating 9.4 10 15 33
materials
Methylene
dichloride 50 75 150 300
Isopropyl alcohol 60 90 180 360
Total (Mg) 276.00 350.00 610.00 1100.00

Mix properly Thiacetazone (0.187 kg) and Isoniazide (0.375 kg) and MCCP (0.100 kg) and shifted through the SS sieve (40 mesh), weigh out the maize starch (0.150 kg) and passed through the SS sieve (100 mesh) and transfer to SS container to rotate for 10 minutes for proper mixing in high speed mixer. In granulation process a hot yellow solution of purified water and PVPk-30 (as binder) added till to form cohesive mass. Granulate the wet granules using suitable device and dry in tray dryer and shift the wet granules through the SS sieve (8 mesh). Dry the granules and shifted through SS sieve (20 Urn) and transfer to milt mill and again shifted though SS sieve (20 mesh) until adequate consistency achieved. Finally mixture of Talcum powder (0.013 kg), Arosil (0.008 kg), Sodium starch glycolate (0.851 kg) and magnesium sterate (0.005 kg) as lubricant shifted through SS sieve (60 mesh) added to granules with proper mixing. Compress the final mixture to form thiaceazone-isoniazide core tablet. Film coat the tablet with an organic preparation consisting of coating materials mixture HPMC, polyethylene glycol, titanium dioxide, quinoline yellow (0.008 kg) in mixture organic solvents such as Methylene dichloride and isopropyl alcohol by using the auto coater to form a film coated tablet.

Claims:
1) A composition for combating mycobacterium tuberculosis using tuberculosis line 1 and tuberculosis line 3 inhibiters, useful for oral administration and capable for pharmaceutical application comprising the steps of:
a) Thiaceatzone or pharmaceutically acceptable salt thereof and;
b) Isoniazide or pharmaceutically acceptable salt thereof and compound of part (a) and part (b) blended together in predetermined weight ratio in film coated oral administered tablet;
c) a tablet core consisting the therapeutically effective combination
of thiaceatzone and isoniazide, preferably in finely powdered having
medium particle size of approximately 100 to 900 µm and optionally
pulverized or micronized with suitable pharmaceutically accepted
excipient(s) such as fillers, binders, lubricants and disintegrants for the
production of granules to the tablet core and to the film coating on
tablet core respectively;

d) coating of suitable hydrophilic film coating pharmaceutically accepted exceipient(s) or materials for outer coating of tablet core and the percentage of coating is 0.1 to 10% with respective to the tablet core;
2) A composition as claim in claim 1, wherein the said predetermined weight ratio, mentioned in claim 1, of said thiaceatzone or pharmaceutically acceptable salt thereof to said isoniazide or pharmaceutically acceptable salt thereof is in range preferably from an about 1:2 to 2:1 respectively.
3) A composition as claim in claim 1, wherein a process for producing the film coated tablet comprises finely powdered blend of thiacetazone and isoniazide with a particle size approximately 100 to 900 µm but preferably size 10 to 500 µm ,with admixture of excipient(s) that are suitable for granulation process, forming active ingredient in to granules, compressing granules to form tablet cores using tableting process and provide the cores for hydrophilic outer coating as a film coating.

4) A composition as claim in claim 1, wherein tablet core consisting
of suitable disintegrants such as polyvinyl pyrrolidone, microcrystalline cellulose, modified starch, sodium starch glycolate and crosscarmellose sodium in 0.1 to 10% weight of tablet core in alone or mixture thereof.
5) A composition as claim in claim 1, wherein tablet core consisting of
suitable binders such as sucrose , lactose, starch, cellulose, microcrystalline
cellulose, hydroxypropyl cellulose, polyethylene glycol and
polyvinylpyrrolidone (PVP k 30) in 0.1 to 10% weight of tablet core in alone
or a mixture thereof.
6) A composition as claim in claim 1, wherein, tablet core consisting of
lubricants such as talc, silica, aerosol, vegetable sterin, sodium starch
glycolate, HPMC and magnesium stearate in alone or a mixture thereof.
7) A composition as claim in claim 1, wherein, tablet core preferably consisting
microcrystalline cellulose as a disintegrent in 0.1 to 5 % weight of tablet core.

8) A composition as claim in claim 1, wherein tablet core preferably consisting of polyvinylpyrrolidione (PVPk30) as a binder in 0.1 to 5% weight of tablet core.
9) A composition as claim in claim 1, wherein tablet core preferably consisting of talc, aerosol, sodium starch glycolate and magnesium sterate as a lubricants in 10-90 % weight of tablet core or in alone or mixture thereof.
10) A composition as claim in claim 1, wherein, the film coating materials
mixture are consisting of HPMC, polyethylene glycol, titanium dioxide and
percentage of coating mixture is preferably 1-3% weight of tablet core.