Classifications

• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
  • A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
  • A61K31/7048—Compounds having saccharide radicals and heterocyclic rings having oxygen as a ring hetero atom, e.g. leucoglucosan, hesperidin, erythromycin, nystatin, digitoxin or digoxin
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K9/00—Medicinal preparations characterised by special physical form
  • A61K9/10—Dispersions; Emulsions
  • A61K9/127—Synthetic bilayered vehicles, e.g. liposomes or liposomes with cholesterol as the only non-phosphatidyl surfactant
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P33/00—Antiparasitic agents
  • A61P33/02—Antiprotozoals, e.g. for leishmaniasis, trichomoniasis, toxoplasmosis
• • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
  • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
  • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
  • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
  • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Definitions

• FIELD OF INVENTION The invention relates to sterol enriched mixed lammelarity 10 Amphotericin intercalating liposomes in saline and the process of preparing it.
• Leishmaniasis is caused by macrophage resident intracellular- protozoan parasites Leishmania donovani, Leishmania infantum and 15 Leishmania chagasi. Parasite transmission occurs through the bite of infected female phlebotomine sandfly. Sandfly obtains infected blood while sucking the blood from their parasite carrier hosts.
• Leishmaniasis is caused by macrophage resident intracellular- protozoan parasites Leishmania donovani, Leishmania infantum and 15 Leishmania chagasi. Parasite transmission occurs through the bite of infected female phlebotomine sandfly. Sandfly obtains infected blood while sucking the blood from their parasite carrier hosts.
• Four different forms of Leishmaniasis with different clinical manifestations occur in humans.
• Visceral Leishmaniasis which is 20 also known as kala-azar is the most severe and has a mortality rate of almost 100% if not treated. Frequent bouts of fever, weight loss, enlargement of spleen and liver accompanied with anemia are characteristic signs and symptoms of Visceral Leishmaniasis.
• Amphotericin B is a polyene antibiotic produced by soil bacteria 25 Streptomyces nodosus. Amphotericin B is known to be the most potent antifungal drug since its discovery in 1950s. Later it was found to be an effective treatment for kala-azar also (Trans Royal Soc. Trop. Med. Hyg. 1963; 57: 266-268).
• Amphotericin B is free of drug resistance problems (CP Thakur et.al. 1993. The National Medical Journal of India; 6: 57-60 and CP Thakur et.al. 1993. Indian Journal of Medical Research; 97: 170-175). Despite being an effective anti-lieshmanial drug, amhotericin B has been used cautiously only as second line drug due to it's infusion related adverse effects and toxicities, predominantly nephrotoxicity, cardio- toxicity and neuro-toxicity.
• amphotericin B Strategic suitability of liposomal formulations of amphotericin B for the treatment of Visceral Lieshmaniasis emanates from several factors including factors such as leishmania resides in the scavenger cells such as macrophages, liposomes are rapidly taken up by the macrophages, amphotericin B is an effective anti-leishmanial drug, and amphotericin B can be formulated as liposomal preparation. All of these facilitate targeted delivery of Liposomal amphotericin B to the desired site of anti-parasitic action and reduce toxicities of amphotericin B.
• liposomes offer an amplification effect through concentrated encapsulation of numerous molecules of drug in each liposome particle, which are delivered at the site of action (Gregoriadis G, 1995. Engineering liposomes for drug delivery: Progress and Problems. TIBTECH; 13: 527-537).
• Prevention of toxicity by encapsulating amphotericin B in liposomes has made liposomal amphotericin B the best option for the treatment of Visceral Leishmaniasis.
• the critical lipid constituent for minimizing amphotericin B toxicity of formulations for use in treatment of systemic mycosis, are sterols.
• amphotericin B constituted of ergosterol with certain phospholipids have been reported to cause lysis of red cells in-vitro and thus believed to be unsuitable (Mehta, R. et.al. 1984, Biochimica et Biophysica Acta; 770: 230-234). Unsuccessful attempts with limited number of phospholipid combinations and ergosterol has prevented further efforts on development of ergosterol containing amphotericin B formulations (New, P.R.C. et.al.1981, J.
• Liposomes are known to be rapidly phagocytosed in macrophages and will obviously result in concentration of liposome encapsulated amphotericin B in the macrophages. To formulate liposomes with higher concentration of amphotericin B without causing toxicity required strategies, which are not yet reported. Understanding of sterol synthesis is important for rational designing of quick, effective and specific treatment of leishmaniasis using liposomal amphotericin B. Lipids account for 15% of the dry weight of the leishmanial cells (Meyer and Ilolz, 1966, J. Biol. Chem. 24: 5000-5007). Metabolism of lipids is crucial lo" "Seve al vital physiological processes and effect parasite's survival.
• Ergosterol or its precursor episterol is synthesized de novo from AcetylCoA to evalonate to Sqalene to Lanosterol followed by four more steps to finally Ergosterol ( Coppens and Courtoy. 1995, Mol. Bichem. Parasitol. 73,179- 188 and Patent No. US 6,403,576B1). It is pertinent to note that Leishmania require cholesterol for their sustenance and fulfill their requirement by salvaging cholesterol from their host macrophage cells. Effective drug designing prudently shall involve adversely effecting survival of Leishmania by depriving Leishmania of their cholesterol requirement.
• the main object of the invention is to provide a sterol enriched mixed lammelarity amphotericin intercalating liposomes for treatment of infections for targeted delivery of liposomal amphotericin B thereby reducing toxicities of amphotericin B.
• Another object of the invention is to make liposomal amphotericin B with suitable replacement of cholesterol.
• Cholesterol is normally used for preventing leakiness of liposomal drugs / vaccines and in case of amphotericin B formulations cholesterol is used for targeting and minimizing drug toxicities. Cholesterol, however, supports leishmania survival inside the host , therefore should not be taken as constituent of formulation. Yet another object of the invention is to deliver higher concentration of amphotericin B to the target area without causing toxicity. Another object of invention is to sonicate liposomal pharmaceuticals before administration to increase the plasma half-life of the liposomal drug for facilitating better bio-distribution in the body.
• Yet another object of the instant invention is to provide a sugar free composition containing liposomal amphotericin B and therefore freely usable by diabetics
• the instant invention provides for sterol enriched mixed lamellarity Amphotericin intercalating liposomes in 0.9% saline wherein the concentration of
• Amphotericin B is 1 to 15 mg/ml for treatment of infections. There is high Amphotericin B concentration in liposomes.
• the sterol is ergosterol with or without cholesterol.
• the sterol are in combination with phospholipid.
• the said phospholipid is phosphatidylcholine.
• the ergosterol constitutes 50% of the total lipid of the liposome.
• One example of the preferred ratio of phosphatidylcholine, ergosterol and amphotorecin B is 5 : 2 : 1. Sonication before administration increases plasma half-life and provides for better bio distribution.
• Choice of lipid constituents of liposomes particularly sterol is critical to prudent designing of potently effective and safe anti- leishmanial drug. Inclusion of appropriate sterol is necessary to prevent binding of liposomal amphotericin B to mammalian host cells. Delivery of cholesterol in macrophage home of Leishmania through the phagocytic uptake of cholesterol containing liposomal amphotericin B negates optimal benefits of targeted delivery of amphotericin B. Another sterol option of ergosterol in combination with certain lipids in liposomes as outlined above has been shown to cause toxicity to red blood cells in-vitro.
• This invention was taken up to design cholesterol free and suitable sterol containing liposomal amphotericin B.
• Ergosterol with or without cholesterol and in combination with phospholipids such as phosphatidylcholine irrespective of their fatty acid compositions from either synthetic or from natural sources such as Soy in a range of ratios was found suitable for formulating liposomal amphotericin B which is safe and potently effective against infections such as Leishmania.
• Amphotericin B concentration range was used between 1-15 mg /ml of the final preparation.
• the lipids i.e.
• phospholipids preferably phosphatidylcholine and ergosterol are taken in such ratios where ergosterol constitutes up to 50 molar % of the total lipids of the liposomal formulations.
• One preferred formulation for example is PC: Ergosterol : Amphotericin B in the molar ratio of 5:2:1 in aqueous medium. Amphotericin B is well documented to precipitate in saline. Amphotericin B in liposomes of phoshopholipid compositions reported herein above are stabilized and no precipitation occurs even on long term storage. The invention permits replacement of dextrose with saline with or without buffers.
• liposomal amphotericin B 0.9 % Sodium Chloride for suspension of liposomal amphotericin B.
• Such liposomal formulations are stable both in suspension or when stored after lyophilization.
• lyophilized Liposomal Amphotericin B may be reconstituted in aqueous solutions such as 0.9% Sodium Chloride.
• the process of preparing the liposomal Amphotericin B is explained but not limited by the following examples.
• Liposomes can be manufactured by employing a variety of methods either alone or . in combination (Szoka, F. Jr. and Papahadjopoulos, D., 1980. Ann. Rev. Biophys. Bioeng. 9: 467-508).
• Example 1 Example 1
• Methanolic solution of the Liposomal constituents are evaporated to give a film of the constituents. This film is hydrated using aqueous solutions and multilamellar Liposomes are produced by sheering of the film.
• Example 2. Methanolic solution of the Liposomal constituents are processed in Spray Dryer and then hydrated and reconstituted in aqueous medium of choice to give mixed lamellar Liposomal preparation.
• Example 3. Methanolic solution of the Liposomal constituents are evaporated to give a film of the constituents, aqueous solution of choice is added and then subjected to Sonication resulting predominantly into Unilamellar Liposomal Preparation.

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• Health & Medical Sciences (AREA)
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• Veterinary Medicine (AREA)
• Pharmacology & Pharmacy (AREA)
• Medicinal Chemistry (AREA)
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• Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
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• Tropical Medicine & Parasitology (AREA)
• Communicable Diseases (AREA)
• Oncology (AREA)
• Medicinal Preparation (AREA)
• Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
• Manufacturing Of Micro-Capsules (AREA)

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• 2005
  • 2005-06-13 CN CNA200580023366XA patent/CN101014322A/zh active Pending
  • 2005-06-13 EP EP05759293A patent/EP1771156B8/en not_active Expired - Lifetime
  • 2005-06-13 AT AT05759293T patent/ATE476965T1/de not_active IP Right Cessation
  • 2005-06-13 JP JP2007526704A patent/JP4966855B2/ja not_active Expired - Fee Related
  • 2005-06-13 BR BRPI0511371-7A patent/BRPI0511371A/pt active Search and Examination
  • 2005-06-13 US US11/570,379 patent/US20070218119A1/en not_active Abandoned
  • 2005-06-13 WO PCT/IN2005/000193 patent/WO2005120460A1/en not_active Ceased
  • 2005-06-13 AU AU2005251572A patent/AU2005251572B2/en not_active Ceased
  • 2005-06-13 DE DE602005022876T patent/DE602005022876D1/de not_active Expired - Lifetime

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