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OA13318A - Microsphere of two anti-tubercular drugs and a biodegradable polymer administered by inhalation alone or combined with oral route, and process thereof. - Google Patents

Microsphere of two anti-tubercular drugs and a biodegradable polymer administered by inhalation alone or combined with oral route, and process thereof. Download PDF

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Publication number
OA13318A
OA13318A OA1200600134A OA1200600134A OA13318A OA 13318 A OA13318 A OA 13318A OA 1200600134 A OA1200600134 A OA 1200600134A OA 1200600134 A OA1200600134 A OA 1200600134A OA 13318 A OA13318 A OA 13318A
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polymer
drug
drugs
tubercular
tuberculosis
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OA1200600134A
Inventor
Suryakumar Jayanthi
Pawan Muttil
Himadri Sen
Rolee Sharma
Rakesh Sinha
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Council Scient Ind Res
Lupin Lmd
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Publication of OA13318A publication Critical patent/OA13318A/en

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    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00Medicinal preparations containing organic active ingredients
    • A61K31/33Heterocyclic compounds
    • A61K31/395Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/4409Non condensed pyridines; Hydrogenated derivatives thereof only substituted in position 4, e.g. isoniazid, iproniazid
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00Medicinal preparations containing organic active ingredients
    • A61K31/33Heterocyclic compounds
    • A61K31/395Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/445Non condensed piperidines, e.g. piperocaine
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00Medicinal preparations characterised by special physical form
    • A61K9/0012Galenical forms characterised by the site of application
    • A61K9/007Pulmonary tract; Aromatherapy
    • A61K9/0073Sprays or powders for inhalation; Aerolised or nebulised preparations generated by other means than thermal energy
    • A61K9/0075Sprays or powders for inhalation; Aerolised or nebulised preparations generated by other means than thermal energy for inhalation via a dry powder inhaler [DPI], e.g. comprising micronized drug mixed with lactose carrier particles
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00Medicinal preparations characterised by special physical form
    • A61K9/14Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
    • A61K9/16Agglomerates; Granulates; Microbeadlets ; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction
    • A61K9/1605Excipients; Inactive ingredients
    • A61K9/1629Organic macromolecular compounds
    • A61K9/1641Organic macromolecular compounds obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyethylene glycol, poloxamers
    • A61K9/1647Polyesters, e.g. poly(lactide-co-glycolide)

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  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Epidemiology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Otolaryngology (AREA)
  • Pulmonology (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Description

01 33 1 fi
Oounoxl of Sciontific anal Industrial Âeaoarab,and Lupin îdaited Kessaroh. Part
MICROSPHERE OF TWO ANTI-TUBERCULAR DRUGS AND A BIODEGRADABLE POLYMERADMINISTERED BY INHALATION ALONE OR COMBINED WITH ORAL ROUTE, AND PROCESSTHEREOF
Technical Field 5 The présent invention relates to a biodégradable microparticle composition useful for tiretarget spécifie drug delivery to manage pulmonary tuberculosi-ô, said compositioncomprising two anti-tuberculosis drugs, and a biodégradable polymer for drug delivery in aratio of about 1:2 to 2:1, wherein the anti - tubercular drugs are in thé ratio of 1:2 to 2:1,also, a process for the préparation of the composition, and lastly, a method of treating 10 pulmonary tuberculosis in a subject, said method comprising administering by inhalationalone or in combination with oral route, pharmaceutically effective amount of thecomposition to the subject in need thereof, wherein the dosage for inhalation is rangingbetween 0.5 to 10 mg/kg body weight/day and that for oral route is ranging between 4 to32 mg/kg body weight/day. 15 Background Art
Most infectious diseases are caused by parasitic microorganisms, which résidé in spécifieareas in the human body. The cells présent in these areas are infected by thesemicroorganisms thereby resulting in the localization of these disease-causing agents tospécifie cells. 20 Macrophages are cells that are part of the human immune system and are présent in theliver, lungs, spleen, lymph nodes, thymus, gut, marrow, brain, connective tissue and serouscavities. [11· Their main functions as a first line defence against infectious, is byphagocystosis of the microorganisms. 121 But certain facultative or obligate intracellularparasites use macrophages as safe haven. In patients infected by these microorganisms, the 25 macrophages act as réservoirs for them. ^3,41.
Tuberculosis (TB) is a leading cause of infectious lung disease, and considered theforemost cause of death due to a single microorganism. Tuberculosis has become asignificant opportunistic disease among population with a high incidence of acquiredimmunodeficiency syndrome (AIDS). 30 The occurrence of TB is most often due to Mycobacterium tuberculosis (MTB) infection,and the lungs are the primary site of infection for the systemic pathogen. Problems createdby bacterial infection are linked to their ability to survive and multiply inside the body,especially in the lungs, and to the natural immune response of the infected host. 1 013318
Tuberculosis, and more importantly pulmonary tuberculosis, is best treaied successfully only through a treatment schedule than can achieve sustained drug concentrations for prolonged periods. But sometimes even such methods results in failure. One reason couid be that oral administration of drugs may not achieve appréciable concentrations in the 5 cytosol of target cells like macrophages.
Bacteria reaching the deep lung are phagocytized by alveolar macrophages in the fîrst stepof pathogenesis, Inside the macrophage the bacteria will either be destroyed, beginreplicating, or remain latent indefînitely. If réplication is not prevented, the bacilli willmultiply and eventually cause the macrophage to rupture. I o Current treatments of tuberculosis are limited by their methods of delivery. Persistent, highblood levels of anti tubercular drugs resulting from prolonged oral administration may notbe sufiicient to kill mycobacteria residing in macrophages.
In order to solve this problem, several investigators hâve proposed the administration ofthe drugs in the form of vesicular Systems as inhalations t5J or injectable préparation as 1S also microparticulate Systems for injection. [7,8\ Recently, O’Hara and Hickey suggestedthat administration of biodégradable microspheres through the bronchio-pulmonary routefor better therapy of tuberculosis.
In order to target the microorganisms présent in the macrophages, it is essential to developa formulation whereby the therapeutic agent can be delivered at high concentration into the 20 macrophage, It is a known fact that even when the drugs are in high concentration insoluble form in the sérum, very little of it reach the macrophages. Moreover, macrophagesare programmed to phagocytose any foreign particle they encounter thereby making itdifBcult to build up high intra cellular concentration of the drug in a macrophage. One wayto overcome this problem is using carriers for transporting the drugs. Liposomes and 25 microparticles are two types of carriers, which hâve been widely studied.
Drug doses above those currently administered would présent the risk of toxic side effectssince the anti-tubercular agents are at their maximum tolerated dose for systemic exposure.Targeting anti-tubercular drug delivery to the lung may increase local therapeutic effectand reduce systemic exposure.
30 Various researchers hâve reported the development of dry powder inhalations forpulmonary delivery of drugs t10·11'121. Particles delivered to the lungs are rapidlyphagocytosed by the alveolar macrophages.[13J 2 013318 US Patent No. 6,264,991 discloses compositions and methods for treating intracellularinfections comprising administefing a first effective amount of a suitable drug contained infirst biocompatible microspheres having a diameter less than 10 microns and a second setof microspheres having a diameter more than 10 microns, to provide continuing systemic 5 release of tlie drug. The administration of the first microspheres is by intravenous route andthe second microspheres by subcutaneous route.
In case of tuberculosis, as the microorganisms résidé in the macrophages, the inhaledmicroparticles hâve a potential to deliver the drug directly into the macrophages resultingin higher concentrations than when given orally. 10 Moreover, this may resuit in the possibility of réduction in dosage amount & frequency orduration oftreatment.
Alveolar macrophages having anti-TB drugs in microparticulate System along withmycobacterium travel through lymphatic circulation to secondary lymphoid organs. Thus,mycobacteria disseminate not only through the bloùdstream, but also to sites where 15 alveolar macrophages traffic. Boading bacteria résident alveolar macrophages with drugcontaining microparticles leads to transportation of drugs to ail the sites where migratingmacrophages go thereby mimicking the course of spread of bacteria. As a resuit sufficientdrug concentration may also be achieved- in the various tissues where bacteria tend tomigrate. Due to the presence of a biodégradable polymer, controlled release of drugs may 20 be obtained, thereby prolonging the duration of action,
Following are the advantages of noninvasive (ïnhalable) biodégradablemicroparticles: a) Targeting the primary site of tuberculosis infection i.e. lungs. b) Targeting mycobacterium infected macrophages. 25 · c) Rate controlled release of drugs using sélective polymer, their ratios andmicroparticulate size. d) Substantial increase of drug concentrations within macrophages, thereby decreasingsystemic exposure Of drugs -resulting in reduced potential side effects.
Tuberculosis treatment being a long-term schedule, these advantages can resuit in greater 30 patient compliance along with better bioavailability.
Pharmaceutical Research Vol-18 No 10, Oct 2001 discloses Rifampicin and Isoniazidcombinations which is known to form adduct which might resuit in reduced bio-availabilityand efficacy of these drugs. Furtherhigh concentration ofpolymers as much as 3 times 3 013318 w/w of drugs hâve been used. The instant invention relates to bio-degradable, inhalablemicroparticles containing compatible anti-tubercular drug combination. As the présentinvention uses lesser quantities of the polymer, the cost of the therapy will also be cheaper.The US Patent No. 6,264,991 refers to the treatment of tuberculosis using one ôr twodrugs. However, the dosage and ratios of the constituents are totally distinct. These change in the dosage, size of the particles, and route of administration are critical for the desired »? , results. Further, the published article of the inventors in Pharmaceuticai Research Vol18,No 10 October 2001 refers to the management of tuberculosis. However, tire drugs usedare distinct from that of the instant Application. The ratio of the polymer and the drug iscritical for the functioning of the composition. Also, ratio of drugs is critical for thedesired results. Most importantly, there is no adduct formation in the case of instantApplication.
Objects of the présent invention:
The main object of the présent invention is to develop an inhalable, biodégradablemicroparticle composition comprising one or more anti-tuberculàr drugs for the targetspécifie drug delivery to manage pulmonary tuberculosis.
Another objective of the présent invention is to provide a method of enhancing the efficacyof anti-tubercular drugs by administering them through inhalation route.
Another main invention of the présent invention is to develop a method of treatingpulmonary tuberculosis in a subject, said method comprising administering by inhalationalone or in combination with oral route, pharmaceutically effective amount of composition,comprising bio-degradable inhalable microparticles useful for the target spécifie drugdelivery to manage Pulmonary tuberculosis containing two anti -tubercular drugs and bio-degradable polymer in the drug delivery in the ratio of 1:2 to 2:1 to the subject in needthereof, wherein the anti - tubercular drugs are in the ratio of 1:2 to 2:1,
Still another object of the présent invention is to develop a method to target alveolar f macrophages.
Still another object of the présent invention is to develop a method of treating tuberculosis,which helps the drug reach the site in high amount consistently.
Still another object of the présent invention is to develop a method to help reduce theamount ofdrug required for the treatment of tuberculosis.
Still another object of the présent invention is to develop a process for the préparation of abiodégradable microparticle composition comprising one or more anti-tubercular drugs,and a biodégradable polymer for drug delivery. 4 013318
Summary of the présent Invention
The présent invention relates to a biodégradable microparticle composition useful for thetarget spécifie drug delivery to manage pulmonary tuberculosis, said compositioncomprising two anti-tuberculosis drugs, and a biodégradable polymer for drug delivery in a 5 ratio of about 1:2 to 2:1, wherein the anti - tubercular drugs are in the ratio of 1:2 to 2:1,also, a process for the préparation of the composition, and lastly, a method of treatingpulmonary tuberculosis in a subject, said method comprising administering by inhalationalone or in combination with oral route, pharmaceutically effective amount of thecomposition to the subject in need thereof, wherein the dosage for inhalation is ranging 10 between 0.5 to 10 mg/kg body weight/day and that for oral route is ranging between 4 to32 mg/kg body weight/day. <·,···
Detailed Description of the présent Invention
Accordingly, the présent invention relates to a biodégradable microparticle compositionuseful for the target spécifie drug delivery to manage pulmonary tuberculosis, said 15 composition comprising one or more anti-tuberculosis drugs, and a biodégradable polymerfor drug delivery in a ratio of about 1:2 to 2:1, wherein the anti - tubercular drugs are inthe ratio of 1:2 to 2:1. Also, the invention relates to a process for the préparation of thecomposition, and lastly, a method of treating pulmonary tuberculosis in a subject, saidmethod comprising administering by inhalation alone or in combination with oral route, 20 pharmaceutically effective amount of the composition to the subject in need thereofwherein the dosage for inhalation is ranging between 0.5 to 10 mg/kg body weight/dayand that for oral route is ranging between 4 to 32 mg/kg body weight/day.
In still another embodiment of the présent invention, wherein a biodégradablemicroparticle composition useful for the target spécifie drug delivery to manage pulmonary 25 tuberculosis, said composition comprising one or more anti-tuberculosis drugs, and abiodégradable polymer for drug delivery.
In still another embodiment of the présent invention, wherein the drugs are in the ratio ofabout 1:1.
In still another embodiment of the présent invention, wherein the ratio of drug to polymer 30 is about 1:1.
In still another embodiment of the présent invention, wherein the drugs are selected from agroup comprising Rifabutin, Rifapentine, Rifampicin, Isoniazid, pyrazinamide andethambutol.. 5 013318
In still another embodiment of the présent invention, wherein the drugs are in preferred combination ofRifabutin and Isoniazid. .
In still another embodiment of the présent invention, wherein the polymer is selected from a group comprising polyglycolic acid, polylactic acid, poly(lactic acid-co-glycolic acid), 5 polysebacic anhydride,and polycaprolactone and mixtures thereof.
In still another embodiment of the présent invention, wherein the preferred polymer ispolylactic acid.
In still another embodiment of the présent invention, wherein the size of the biodégradablemicroparticles is ranging between 1-15 micron. 10 In still another embodiment of the présent invention, wherein at least 90% of themicroparticles are of size less than 10 microns.
In still another embodiment of the présent invention, wherein a method of treatingpulmonary tuberculosis in a subject, said method comprising administering by inhalationalone or in combination with oral route, pharmaceutiçally effective amount of composition 15 comprising of bio-degradable inhalable microparticles useful for the target spécifie drugdelivery to manage Pulmonary tuberculosis dontaining one or more anti -tubercular drugsand bio-degradable polymer in the drug delivery in the ratio of 1:2 to 2:1 to the subject inneed thereof, wherein the anti - tubercular drugs are in the ratio of 1:2 to 2:1.
In still another embodiment of the présent invention, wherein the said method targets 20 alveolar macrophages.
In still another embodiment of the présent invention, wherein said method shows nodeleterious effect on the subject.
In still another embodiment of the présent invention, wherein said subject is an animalincluding humans. 25 In still another embodiment of the présent invention, wherein the dosage for inhalation is• ranging between 0.5 to 10 mg/kg body weight/day.
In still another embodiment of the présent invention, wherein the dosage for oraladministration is ranging between 4 to 32 mg/kg body weight/day.
In still another embodiment of the présent invention, wherein said method helps drug reach 30 the site in high amount consistently.
In still another embodiment of the présent invention, wherein said method helps reducesthe amount of drug required for the treatment of tuberculosis. 6 013318
In still another embodiment of the présent invention, wherein a process for the préparationof a' microparticle composition comprising one or more anti-tuberculosis drugs, and abiodégradable polymer for drug delivery, said process comprising steps of; • dissolving the drug (s) in an aqueous solvent preferably, alcohol or any othersuitable solvent to obtain a solution, • dissolving the polymer in dichloromethane or any other suitable solvent to obtainanother solution, • mixing the aforementioned solutions to obtain the final solution, and • spray-drying the final solution to obtain the microparticles.
In still another embodiment of the présent invention, wherein the microparticles are of thesize ranging between 1-15 micron.
In still another embodiment of the présent invention, wherein at least 90% of themicroparticles are of size less than 10 microns.
In still another embodiment of the présent invention, wherein the drugs are selected from agroup comprising Rifabutin, Rifapentine, Rifampicin,. Isoniazid, pyrazinamide andethambutol..
In still another embodiment of the présent invention, wherein the polymer is selected froma group comprising polyglycolic acid, polylactic acid, poly(lactic acid-co-glycolic acid),polysebacic anhydride, polycaprolactone or mixtures thereof.
Brief description of the présent Invention
Figure 1 shows the results after treatment of tuberculosis with the composition of theinstant Application for 3 weeks.
Figure 2 shows the results after treatment of tuberculosis with the composition of theinstant Application for 4 weeks.
Typically anti-tubercuiar drugs such as Rifampicin, Isoniazid, Ethambutol andPyrazinamide are given orally in the range between 4- 32 mg / kg body weight / day.
Table 1: Lists the dosage ranges of the anti-TB drugs per day (Lower dosage is for a subject of body weight less than 50 kgs) ΑΝΤΙ -TB DRUGS ORAL DOSE (WHO APPROVED) PER KG BODY WT. 1. RIFAMPICIN - 450 mg to 600 mg / day Range for ail drugs isbetween 4 to 32 mg/kg bodyweight/day 2. ISONIAZID - 225 mg to 300 mg / day 3. ETHAMBUTOL - 825 mg to 1100 mg / day 4. PYRA ZINAMIDE - 1200 mg to 1600 mg/day 7 013318
Table 2: Lists the dosage ranges of the anti-TB drugs on the basis of per kg bodyweight ANTI-TB DRUGS ON THE BASIS OF PER KG BODY WT. 1. RIFAMPICIN -9 to 12 mg/ kg body weight 2. ISONIAZID - 4.5 to 6 mg/ kg body weight 3. ETHANBUTOL - 16.5 to 22 mg/ kg body weight 4. PYRAZINANIDE - 24 to 32 mg/ kg body 5. RIFABUTIN - 300 mg/day is équivalent to 6 mg/kg body weight
One aspect of the invention provides a method for the préparation of microparticles of oneor more therapeutic agents along with a biodégradable polymer. 5 In a preferred embodiment the microparticles contain one or more anti-tubercular drugs. Ina preferred embodiment, the anti-tubercular drugs are selectedfrom the group consisting ofRifampicin, Rifabutin, Rifapentïïie,' Isoniazid, pyrazinamide and ethambutol.
In another preferred embodiment the ratio of Rifabutin to Isoniazid is 1:2 to 2:1 and thedrugs to polymer ratio is also 1: 2 to 2 :1. 10 In a preferred embodiment, the biodégradable polymer is selected from the groupcomprising polyglycolic acid, polylactic acid, polyflactic acid-co-glycolic acid)polysebacic anhydride and polycaprolactone or mixtures thereof . In another preferredembodiment, the biodégradable polymer is polylactic acid.
The composition of the invention may be formulated using techniques generally known in 15 the art of microparticle synthesis. In a preferred embodiment, the microparticles are madeby the spray - drying technique. In one particular embodiment, the microparticles hâve asize ranging from 1 to 15 microns with at least 90% of the microparticles having a size 'below 10 microns.
To accomplish the objects of the présent invention, the microparticles can be' administered 20 in inhalable form for pulmonary dêlivery. Tfie methods and composition of this inventioncan be used to treat pulmonary tuberculosis in any animal.
Moreover, the invention also describes methods of combining two altemate routes of drugadministration to achieve signifîcant réduction in bacterial levels as compared toconventional therapy. 25 Methods for making spécifie and preferred compositions of the présent invention aredescribed in the example below. However, these examples should not be construed to liraitthe scope of the invention. 8 013318
Préparation of Rifabutin + Isoniazid microparticles
Isoniazid (750 mg) was dissolved in methanol (8 mL) by warming lightly. Poly-lactic acid(1.5 g) and Rifabutin (750 mg) was dissolved in dichloromethane (90 ml). Both solutionswere mixed using 2 mL portions of methanol to wash the Isoniazid solution. The solutionwas spray dried using Buchi Spray Dryer. The solution feed rate was adjusted to 5mL Zminute at a setting of 13% with the inlet température of 52°C and aspirator at 70%. Theatomizing air was supplied at l.ôkgZcm2. The microparticles were analysed for the drug(s)content and the particle size was determined using Malvem Particle sïze Analyzer ModelMastersizer 2000.
Example No. 1
In vivo. Studies on Mice:
Préparation of Microparticles / Drug Solutions i a. Rifabutin + Isoniazid Microparticles ·
The above microparticles were prepared as described earlier. Particle size analysis of themicroparticles showed 90% below 10 microns. b. Rifabutin solution t r.· , ‘
Rifabutin solution was prepared for oral administration using 10% DMSO in water. c. Isoniazid solution
Isoniazid solution was prepared for oral administration using water.
Infection &amp; Treatinent:
Swiss Albino Mice 4-6 weeks old weighing 18-22 gms were used for the study. The micewere infected with 107 CFUs /0.2 mL of Mycobacterium tuberculosis H37RV strain byintravenous route. Ail infected animais were distributed in 8 groups of six mice each.Infected mice were treated with the drug formulations 24 hours post infection as belôw:Group 1- Rifabutin + Isoniazid microparticle by inhalation (15-20 mg / 30 sec /mouse)Group 2- Rifabutin + Isoniazid microparticle by inhalation (15-20 mg / 30 sec /mouse)along with oral administration of free Rifabutin (5 mg / kg) and free Isoniazid (5mg / kg)Group 3- Oral administration office Rifabutin (10 mg /kg) and free Isoniazid (10 mg /kg)Group 4- Oral administration of free Isoniazid (25mg/kg)
Group 5- Oral administration of free Rifabutin (20mg/kg)
Group 6-Early infection control
Group 7- Late infection control
Group 8- Négative (V ehicle control) 9 013318
Treatment Schedule
Ail animais were treated once a day, 5 days per week for 3 weeks / 4 weeks.
In groups treated by inhalation, the microparticles were delivered by generating aérosols ina tube witli a rubber teat.
Early Controls were sacrificed 24-hours post infection i.e. at the start of the treatment.
Ail animais treated as well as untreated late Controls were sacrificed 3 days post treatmentto enumerate the mycobacterial load in the organs. Viable tubercle bacilli from the targetorgans i.e, lung and spleen wêre enumerated by plating out the different dilution onindividual organ homogenates on the Middle brook 7H 10 media plàtes. The plates wereincubated at 30°C for 3-4 weeks. Mycobacterial load in each organ was determined bycounting the colonies on the plates.
Observation:
After treatment for 3 Weeks
In the présent study no growth was seen in Iungs of 4 out of 6 animais when theformulation was administered by inhalation route alone (Group 1). The spleens of theanimais however showed the presence of low levels of viable bacilli.
Lungs and spleen were found to be free of infection in ail the mice treated by inhalation incombination with half the oral dose of pure drugs (Group 2). Significant réduction (2 log)in the bacterial load was observed in both the organs (lung, spleen) in the animais treatedby oral route with either combination of pure drug or alone.
After treatment for 4 Weeks
Lungs in ail the mice were found to be free of infection when treated by inhalation routealone (Group 1). The spleens of the animais however showed the presence of low levels ofviable bacilli.
Lungs and spleen were found to be free of infection in ail the mice treated ’by inhalation incombination with half the oral dose of pure drugs(Group 2), Significant réduction (2 log)in the bacterial load was observed in both the organs (lung, spleen) in the animais treatedby oral route with either combination of pure drug or alone.
The results indicate that treatment of M.tuberculosis infected animais by inhalation routeusing microparticles in combination with oral route (half the dose) results in fasterclearance of tubercle bacilli from the organs.
Figures 1 &amp; 2 show the results after treatment for 3 and 4 weeks respectively. 10 013318
Normally ail the 4 Anti-Tubercular drugs (Rifampicin, Isoniazid, Ethambutol and
Pyrazinamide) are admïnistered to tuberculoses patients for a period of at least 2 months, before switching over to Rifampicin and Isoniazid for atleast 4 months. In the présent invention, lungs and spleen were found to be free of infection in ail the mice by using 2 5 drugs only (Rifabutin and Isoniazid),
Further, the Iesser is the number of drugs in the composition, better is the performance.Bibliography 1. B.Vemon-Robert, The macrophage, in : RJ. Harrison, R.M.H. Mcminn (Eds.),Biological structure and fuhction, Vol 2, Cambridge University Press, Condon, 10 .. 1972. 2. D.P. Speert, in : C.E. Lewis, J.O’D. McGee (Eds.)., The Natural Immune System:The Macrophage, Oxford University Press, New York, 1992, PP. 215-263. 3. J.Stewart, D.M.Weir, Immunity in Bacterial infections, in : D. Greenwood,R.Slack, J. Pentherer (Eds.), 14ή Edition, Medical Microbiology, Churchill 15 Livingstrong, Edinburgh, 1942, PP 195-200. 4. G.R. Donowitz. Tissue-directed antibiotics· and intracellular parasites: Complexinteraction of phagocyte, pathogens and drugs, Clin. Infect. Dis. (1994) 926-930. 5. ΥΓΝ. Kurunov, P.A, Filimonov, A.V. Svistelnik et al. Efficacy of liposomizedantibacterial drugs in. inhalation therapy of experimental tuberculosis. Probl. 20 Tuberk 1:38-40 (1995). 6. P,Deol, G.K.. Khuller, and K.Joshi. Therapeutic effîcacies of isohazid andrifampin encapsulated in lung-specific stealth liposomes against mycobacteriumtuberculosis infection induced in mice. Anthnicrobial agents chemother 41:1211-1214 (1997) 25 7. D.C. Quenelle, J.K St as, G.A.Winchester et. al. Efficacy of microencapsulated t·; , rifampin in mycobacterium tuberculosis - infected mice. Antimicrobial AgentsChemother. 43:1144-1151 (1999) 8. E.L. Barrow, G.A. Winchester, J.K. Staas et.al. Use of microsphere technology fortargeted delivery of rifampin to mycobacterium tuberculosis- infected 30 macrophages. Antimicrobial Agents Chemother. 42:2682-2689. 9. P. O’Hara and AJ. Hickey. Respirable PLGA microspheres contaming rifampicinfor the treatment of tuberculosis : manufacture and characterization. Pharm. Res.17:955- 961 (2000) 11 33 1 g 10. Y. Kawashima, T. Serigano, T.Hino et.al. surface-modified antiasthmatic drugpowder aérosols inhaled intratracheally reduce the pharmacologically effectivedose. Pharm. Res. 15:1753-1759 (1998). 11. J.S. Patton. Deep-lung ’delivery of proteins. Modem Drug Discover 2: 19-28 5 (1999) 12.. R..J. Malcolmson and J.K. Embleton. Dry powder formulations for pulmanory delivery. Pharma. Sci.Tech.index 1: 394-398 (1998). 13. C.Evora, I .Soriano, R.A. Rogers et.al. Relating the phagocytosis of microparticlesby alveolar macrophages to surface chemistry: The effect of 1,2-dipalmitoyl 10 phosphatidylcholine. J. Control. Release 51:43-152 (1998). 12

Claims (19)

  1. 013318 Claims:
    1. A biodégradable inhalable microparticle composition useful for the target spécifie drugdelivery to manage pulmonary tuberculosis, said composition comprising two anti-tubercular drugs selected from a group consisting of Rifabutin, Rifapentine, Isoniazid,pyrazinamide and ethambutol and a biodégradable polymer for drug delivery wherein the 5 . ratio of drug (s): polymer is from about 1: 2 to 2: 1.
  2. 2. The microparticle composition as claimed in claim 1, wherein two anti- tubercular drugsare used in the ratio of about 1: 2 to 2: 1.
  3. 3. The microparticle composition as claimed in claim 2, wherein the drugs are in preferredcombination of Rifabutin and Isoniazid.
  4. 4. The microparticle composition as claimed in claim 1, wherein the polymer is selected from a group comprising polyglycolic acid, polylactic acid, poly (lactic acid-co-glycolicacid), polysebacic anhydride, polycaprolactone or mixtures thereof.
  5. 5. The microparticle composition as claimed in claim 1, wherein the preferred polymer ispolylactic acid.
  6. 6. The microparticle composition as claimed in claim 1, wherein the size of the microparticles is ranging between 1-15 micron.
  7. 7. The microparticle composition as claimed in claim 8, wherein at least 90% of themicroparticles are of size less than 10 microns.
  8. 8. Use of a composition of claim 1, in the manufacture of a médicament for treating 20 pulmonary tuberculosis in a subject.
  9. 9. The use as claimed in claim 9, wherein the said médicament is for targeting alveolarmacrophages.
  10. 10. The use as claimed in claim 9, wherein said subject is an animal including humans.
  11. 11. The use as claimed in claim 9, wherein the dosage for inhalation is ranging between 0.5 25 to lOmg/kgbody -13- 013318
  12. 12. The use as claimed in claim 9, wherein the dosage for oral administration is rangingbetween 4 to 32 mg/kg body
  13. 13. The use as claimed in claim 9, wherein said use helps the drug to reach the site in highamount consistently.
  14. 14. The use as claimed in claim 9, wherein said use helps to reduce the amount of drug required for the treatment of tuberculosis.
  15. 15. The use as claimed in claim 9, wherein said use shows 2 log réduction in the bacterialload.
  16. 16. A process for the préparation of a biodégradable microparticle composition comprisingtwo anti-tubercular drugs selected ffom a group consisting of Rifabutin, Rifapentine,Isoniazid, pyrazinamide and ethambutol and a biodégradable polymer for drug delivery,said process comprising steps of; a. dissolving one anti-tubercular drug into an aqueous solvent to obtain a solution, b. dissolving the polymer and other anti-tubercular drug into dichloromethane to obtainanother solution, c. mixing the aforementioned solutions to obtain the final solution, and d. spray-drying the final solution to obtain the biodégradable microparticles.
  17. 17. A process as claimed in claim 16, wherein the microparticles are of the size rangingbetween 1-15 micron. 1S. A process as claimed in claim 16, wherein the aqueous solvent is preferably alcohol.
  18. 19. A process as claimed in claim wherein at least of the microparticles are of size less than 10 microns.
  19. 20. A process as claimed in claim 16, wherein the polymer is selected from a groupcomprising polyglycolic acid, polylactic acid, poly (lactic acid-co- glycolic acid),polysebacic anhydride, polycaprolactone or mixtures thereof. -14-
OA1200600134A 2003-10-22 2003-10-22 Microsphere of two anti-tubercular drugs and a biodegradable polymer administered by inhalation alone or combined with oral route, and process thereof. OA13318A (en)

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