KR100754352B1 - The method of preparing emulsion vehicle for poorly soluble drugs - Google Patents

Abstract

Disclosed are emulsions as excipients or carriers for therapeutic agents, including one or more tocols, co-solvents and stabilized by biocompatible surfactants, which are substantially free of ethanol and can be administered to animals or humans through various routes. Also included in the emulsion is PEGylated Vitamin E. PEGylated α-tocopherols comprise polyethylene glycol subunits attached by succinic acid diesters in the cyclic hydroxyl of vitamin E and act as primary surfactants, stabilizers and secondary solvents in tocol emulsions.

Tocopherol, polyethylene glycol, emulsion, tumor, TPGS

Description

The manufacturing method of the emulsion excipient for poorly soluble drugs {THE METHOD OF PREPARING EMULSION VEHICLE FOR POORLY SOLUBLE DRUGS}

The present invention relates to the field of pharmaceuticals. In particular, the present invention relates to a medicament wherein one or more tocols are used as the primary solvent.

Hundreds of medically useful compounds are discovered each year, but clinical use of these drugs is only possible if drug delivery excipients are developed to carry them as therapeutic targets in the human body. This problem is particularly important for drugs that are required to be intravenously injected or dosed to reach their therapeutic targets but are water insoluble or poorly water soluble. For these hydrophobic compounds, direct injection can be impossible or very dangerous and can cause hemolysis, phlebitis, hypersensitivity, organ destruction and / or death. Such compounds are termed "lipophilic", "hydrophobic", or "amphiphobic" in the most complex form by pharmacists.

A few examples of therapeutic agents in this category are ibuprofen, diazepam, griseofulvin, cyclosporin, cortisone, proleukin, etoposide and paclitaxel. Kagkadis, KA et al. (1996) PDA J Pharm Sci Tech 50 (5): 317323; Dardel, O. 1976. Anaesth Scand 20: 221-24. Sweetana, S and MJU Akers. (1996) PDA J Pharm Sci Tech 50 (5): 330-342.                 

For drugs that cannot be formulated as an aqueous solution, emulsions are typically the most economical and appropriate for administration, although they have the difficult problem of making them sterile and endotoxin so that they can be administered by intravenous injection. The oils used for pharmaceutical emulsions are typically saponifiable oils from triglyceride groups such as soybean oil, sesame seed oil, cottonseed oil, safflower oil and the like. Hansrani, PK et al., (1983) J. Parenter Sci. Technol 37: 145-150. One or more surfactants are used to stabilize the emulsion, and excipients are added to give the emulsion more bioaffinity, stability and less toxicity. Lecithin from egg yolks or soybeans is a commonly used surfactant. Aseptic preparation can be achieved by completely sterilizing all components prior to preparation and then by non-infectious treatment at all stages of manufacture. However, improved ease of manufacture and guarantee of sterility are obtained by the final aseptic treatment by heating or filtration after hygienic preparation. Unfortunately, not all emulsions are suitable for heating or filtration.

Stability is known to be affected by the size and homogeneity of the emulsion. Preferred emulsions consist of suspensions of less than 1 micron having an average droplet diameter of 200 nanometers or less. Stable dispersion of this size range is not easily obtained, but has the advantage that longer circulation in the bloodstream is expected. Moreover, the lack of stable dispersion of this size range results in nonspecific phagocytosis by the reticulocyte system. As a result, the drug is probably more likely to reach its target. Thus, preferred drug emulsions will be designed to be actively ingested by target cells or organs and target away from the RES.

The use of vitamin E in emulsions is known. Injectable oils containing small amounts of vitamin E (e.g., less than 1%, Lyons, RT, Pharm Res 13 (9): S-226, (1996) "lipophilic antitoxins α-tocopherol and β-carotene- With the numerous examples used as antioxidants in emulsions), the first primitive injectable vitamin E emulsions themselves are used for dietary supplementation in sheep and for vitamin E and Hidiroglou was produced for the study of the pharmacokinetics of its derivatives. Hidiroglou M. and Karpinski K. (1998) Brit J Nutrit 59: 509-518.

Vitamin E in the injectable form for mice was prepared by Kato and colleagues. Kato Y., et al. (1993) Chem Pharm Bull 41 (3): 599-604. The micelle solution was prepared with Tween 80, Brij 58 and HCO-60. Isopropanol was used as cosolvent and then removed by vacuum evaporation: the residual oil glass was then vortexed and absorbed in water as a micelle suspension. Emulsions were also prepared by dissolving vitamin E using soybean phosphatidylcholine (lecithin) and soybean oil. Water was added and the emulsion was prepared using sonication.

In 1983, vitamin E emulsion E-Ferol was introduced for vitamin E supplementation and therapy in newborns. Alade S.L. et al. (1986) Pediatrics 77 (4): 593-597. Over the course of several months, 30 children died as a result of the administration of this product, which was immediately recovered by FDA. The surfactant mixture used for E-Ferol to emulsify 25 mg / mL consisted of 9% Tween 80 and 1% Tween 20. After all, the use of these surfactants seems to have caused the unfortunate death. This experience has necessitated the importance of selecting improved formulations and appropriate biocompatible surfactants and carefully monitoring their amounts in parenteral emulsions.

An alternative means of dissolving poorly water soluble compounds is direct dissolution in non-aqueous environments, for example in alcohol (such as ethanol) dimethyl sulfoxide or triacetin. One example in the PCT application WO 95/11039 illustrates the use of vitamin E and vitamin E derivative TPGS in a combination of ethanol and the immunosuppressive molecule cyclosporin. U.S. Patent 5,689,846 discloses various alcohol solutions of paclitaxel. US Pat. No. 5,573,781 discloses the degradation of paclitaxel in ethanol, butanol and hexanol and an increase in the antitumor activity of paclitaxel when delivered in butanol and hexanol relative to ethanol. Alcohol-containing solutions can be administered with caution, but are typically administered intravenously in order to avoid pain, vascular irritation and toxicity associated with bolus injection of these solutions.

U.S. Patent 4,439,432 discloses the preparation of high concentrations of progesterone solutions in tocopherols. Emulsions can be prepared from these solutions for use as a skin treatment for systemic progesterone deficiency, to treat local skin disorders such as psoriasis or for vaginal application. This solution can also be prepared in capsules for oral administration.

European patent application 001,851 (Akzo N.V.) discloses steroid high concentration solutions of the oetran, androstane, and (19-nor-) pregnan series, including tocol and tocol derivatives that are liquid at normal temperatures.                 

PCT publication WO 95/21217 (Dumex Ltd) discloses that tocopherols can be used in particular for the preparation of topical preparations as solvents and / or emulsifiers of drugs which are substantially insoluble in water. The use of vitamin E-TPGS as an emulsifier in formulations containing high levels of α-tocopherol is mentioned in the specification (pages 7-8 and 12). Examples 1-5 disclose topical formulations comprising vitamin E-TPGS in amounts of less than 25% w / w of the preparation as lipid layers (α-tocopherol), drugs and emulsifiers.

PCT publication WO 97/03651 (Danbiosyst UK Ltd) discloses a lipid drug delivery composition comprising at least five components: a therapeutic drug, vitamin E, an oil in which the drug and vitamin E are dissolved, a stabilizer (phospholipid, lecithin or poly Poloxamer, an oxyethylene-polyoxypropylene copolymer) and water. The disclosed therapeutic agents are itraconazole and paclitaxel. A "therapeutic emulsion" composition requires two oils in the disperse phase in which the therapeutic drug is present, other oils, typically triglycerides, such as vitamin E and soybean oil. Example 16, the only working example using packletaxel, also contains both vitamin E and soybean oil.

WO 97/03651 also discloses that tocol derivatives having vitamin E activity and tocotrienols are considered to be the definition of “vitamin E” as used in the publication.

PCT publication WO 97/22358 (Sherman) discloses a microemulsion preconcentrate of cyclosporin dissolved in a solvent system that may include a hydrophilic component selected from tocols, tocopherols, tocotrienols, and derivatives thereof. In addition to α-tocopherol, the publication also refers to β-, δ- and γ-tocopherol, and α-, β-, δ- and γ-tocotrienol or mixtures thereof. These compositions also include a hydrophilic solvent, preferably polyethylene glycol or propylene carbonate, having an average molecular weight of less than 1000. Sherman's second PCT publication (WO 98/30204) discloses a microemulsion preconcentrate of cyclosporin, wherein the solvent system can comprise two hydrophobic solvents, one of which is tocol, tocopherol, tocotrienol and their Selected from derivatives.

N-methyl-2-pyrrolidone (NMP) under the name Pharmosolve ^™ can be used to improve the solubility of poorly soluble drugs in pharmaceutical preparations, which may be used for future use in veterinary medicine with application in humans. Appeared in recent literature. Moreover, polyvinylpyrrolidone (PVP) under the name Povidone ^{™ having} a molecular weight between 2,500 and 100,000 at a concentration of 1 to 5 percent (w / v) of aqueous injectable bases can be used as a co-release with NMP. . U.S. Patent 5,726,181 discloses a suspension comprising an antitumor composition and a highly lipophilic camptothecin derivative with NMP.

); Science and Applications, Taxol

중 p.244, 조제물, 활성도 및 약물동태학.(M. Suffness ed.), CRC Press, New York, 1995). PEG-400의 사용은 파클리탁셀에 한정되지 않으며 폴리에틸렌 글리콜에서의 양호한 용해성을 나타내는 다른 치료제(예를 들어 Etoposide)에 적용될 수 있다. 폴리에틸렌 글리콜 유도체를 포함하는 파클리탁셀의 유도체 형태는 미국 특허 5,614,549에 기술되어 있다. Polyethyleneglycol (PEG) and PVP are examples of two water soluble polymers that are often used to alter the solubility behavior of drugs, including paclitaxel. Although the solubility of paclitaxel is relatively high in both solvents, the solubility of the drug in dilute aqueous solutions suitable for parenteral administration is low and the drug is likely to precipitate during dilution. In a mixture of water containing PEG 400 and 50-100% PEG 400, the solubility of paclitaxel varies from 0.2 to 175 mg / ml, respectively. Thus, paclitaxel solubility is considerably lower when large amounts of water are used, for example 0.03 mg / ml and <0.3 mg / ml, respectively, for 35% PEG 400 and 30% PVP in water. "Solubility of Paclitaxel in a Polyethylene Glycol 400 / Water Mixture" (Straubinger, RM Biopharmacuitics of paclitaxcel (Taxol

); Science and Applications, Taxol

P.244, Formulations, Activity and Pharmacokinetics (M. Suffness ed.), CRC Press, New York, 1995). The use of PEG-400 is not limited to paclitaxel and can be applied to other therapeutic agents (eg Etoposide) that exhibit good solubility in polyethylene glycol. Derivative forms of paclitaxel, including polyethylene glycol derivatives, are described in US Pat. No. 5,614,549.

In addition to poor solubility and surfactants and the likelihood of precipitation with pharmaceutical compositions of drugs in non-aqueous solvents such as alcohols (ethanol, isopropanol, benzyl alcohol, etc.), another problem is that these solvents are toxic, eg from their containers. For example, to extract the plasticizer. For example, the formulations for the currently available anticancer drug paclitaxel consist of a mixture of hydroxylated castor oil and ethanol, and the plasticizer, such as di- (2-ethylhexyl) -phthalate, can be removed from a commonly used intravenous infusion tube and bag. Extract quickly. Harmful reactions to plasticizers, such as dyspnea, have been reported, requiring the use of special injection systems that require additional cost and time. Waugh, et al. (1991) Am J. Hosp. Pharmacists 48: 1520.

In view of these problems, ideal emulsion excipients are inexpensive, nonirritating or even influential and essentially intrinsic, can be finally sterilized by heat or filtration, are stable for at least one year under controlled storage conditions, It can be seen that it accommodates a wide range of water insoluble and poorly soluble drugs and is substantially free of ethanol. In addition to drugs that are lipophilic and soluble in oil, excipients are needed to stabilize drugs that are poorly soluble in lipids and water in the form of emulsions.

Summary of the Invention

To meet these needs, the present invention relates to pharmaceutical compositions comprising: mixtures and therapeutic agents of surfactants, including one or more tocols, surfactants or cosolvents with or without an aqueous phase. The compositions of the present invention may be in emulsion form, ie micelle solution or self-emulsifying drug delivery system. The tocol (or tocopherol) molecule is preferably α-tocopherol. The composition of the present invention is generally substantially free of any monohydric alcohol.

The cosolvent may comprise a polyvinylpyrrolidone with or without a water soluble polymer, preferably polyethylene glycol or N-methyl-2-pyrrolidone. Polyethylene glycol (PEG) having a molecular weight of 100 to 10,000 is the most preferred cosolvent. Most preferred is PEG-400 in an amount greater than 1% by weight relative to the formulation.

Pharmaceutical compositions can be stabilized by the addition of various amphoteric molecules, including anionic, nonionic, cationic, and zwitterionic surfactants. Preferably, these molecules are PEGylated surfactants and optionally PEGylated α-tocopherols.

Amphiphilic molecules include ascorbyl-6-palmitate; Stearylamine; Surfactants such as sucrose fatty acid esters, PEGylated phospholipids, various tocol derivatives and polyoxypropylene-polyoxyethylene glycol nonionic block copolymers. Useful surfactants also include poloxamers, tetronics, TPGS, glutamyl stearate, pegylated mono- and diglycerides, propylene glycol mono- / diesters, polyglyceryl esters, Soutol HS-15, phospholipids, lecithin, fe Lengthened phospholipids, PEGylated sterols, PEGylated cholesterol, and other tocol esters, sucrose esters, fatty acids, bile acids and conjugate bile acids, nonionic and anionic surfactants.

Therapeutic agents include chemotherapy, antibiotics (antiviral, antibacterial, antiparasitic, antimalarial, antifungal), analgesics, antidepressants, antipsychotics, hormones, steroids, angiogenics, inhibitors of angiogenesis Mesine, cytokine.

A more understandable, but not limited, list of drug forms suitable for use in the present invention is listed in PCT application WO 98/30205.

Tocol-based emulsions are particularly advantageous for synergistic biological and therapeutic effects when used to deliver cardiovascular or cancer therapeutics.

An added advantage of particulate emulsions for the delivery of chemotherapeutic agents is the broad nature of the surfactants used in emulsions to overcome multidrug resistance by inhibiting the membrane-bound drug delivery, P-glycoprotein.

The emulsion of the present invention may comprise an aqueous medium when in the form of an emulsion or micelle solution. This medium may contain various additives to help stabilize the emulsion and impart biocompatibility to the formulation.                 

In one form, the present invention relates to a pharmaceutical composition consisting of α-tocopherol, a chemotherapeutic agent selected from taxoids, taxins and taxanes, water and D-α-tocopherol polyethylene glycol 1000 succinate. In another form, the invention relates to a pharmaceutical composition wherein the α-tocopherol, cosolvent, one or more surfactants, water phase and composition are in the form of an emulsion or micelle solution and the solution comprises a therapeutic agent substantially free of monohydric alcohol.

In a preferred form, the cosolvent may be polyethylene glycol, N-methyl-2-pyrrolidone, polyvinyl-pyrrolidone or mixtures thereof.

In a preferred form the surfactant is an α-tocopherol derivative and the polyethylene glycol has a molecular weight of 100 to 10,000 most preferably about 200 to 1000.

In a preferred form the therapeutic agent is a chemotherapeutic agent selected from taxoids, taxins and taxanes.

Pharmaceutical compositions of the invention typically dissolve the therapeutic agent in a co-solvent to form a therapeutic solution; One or more tocols are then formed together with one or more surfactants to the therapeutic solution to form an oil solution of the therapeutic agent in a hydrophilic cosolvent. The oil solution is then mixed with the water phase to form a pre-emulsion. For IV delivery the pre-emulsion is more homogenized to form a fine emulsion. For oral delivery, the oil solution of the therapeutic agent in the co-solvent with the surfactant is typically wrapped in gelatin capsules.

In a preferred form of the method of the invention the therapeutic agent is dissolved directly in polyethylene glycol or tocol oil, which avoids the use of monohydric alcohol as solvent.                 

Detailed description of the invention

The following definitions are provided for a thorough understanding of the present invention:

Tocopherol: Tocopherol is a family of natural or synthetic compounds, also known as tocol or vitamin E. α-tocopherol is the most abundant and active form of this class of compounds and has the following chemical structure (Formula I):

The molecule contains three structural elements, a Chromman head with phenol alcohols and a phytyl tail. Not all tocopherol isomers have three methyl groups on the chromaman head. The simplest isomer does not contain a methyl group on the croman ring, i.e. 6-hydroxy-2-methyl-2-pitylchroman, and in the following uses the term "tocols" to refer to a broader class of compounds. But this is often referred to simply as "token." Other members of this class include α-, β-, γ-, and δ-tocopherol and α-tocopherol derivatives such as tocopherol acetate, phosphate, succinate, nicotinate, and linoleate. In addition to using them as primary solvents, tocopherols and derivatives thereof are useful as therapeutic agents.

Toko phenol 3,7,11 tree - except that with the yen tail has a structure associated with the tocopherol. The structure of d-alpha-tocophenol is shown in formula (II).

Again, as in the case of tocopherol, not all tocophenol isomers have three methyl groups on the Chromaman head. There are four main tocotrienols: α-, β-, γ-, and δ-tocotrienols. Analogous compounds with fewer methyl groups, such as desmethyltocotrienol and didesmethyltocotrienol, are included within the definition of tocotrienols.

Members of the tocopherol family can provide additional benefits and perform specific biological functions.

In addition, the use of these tocopherols allows the emulsion to have a low viscosity and thus is more easily produced and / or processed.

Preferred members of this group include, but are not limited to, d-γ- and d-δ-tocopherols, d-β-, d-δ- and d-γ-tocotrienols.

Tocol : Since all tocopherols and tocotrienols are basically the simplest tocopherols, i.e., derivatives of 6-hydroxy-2-methyl-2-phytylchroman (often referred to as "tocols"), "tocols" has a broad meaning here. It is used to represent tocopherols and tocophenols and their derivative families. Tocols also include tocopherol or tocopherol derivatives, and their common derivatives esterified at 6-hydroxyl on the chroman ring.

Surfactant : A surface active class of amphoteric molecules prepared by chemical methods or purified from natural sources or methods. These may be anionic, cationic, nonionic, and zwitterionic. Typical surfactants are described in Emulsions: Theory and Practice, published by Paul Becher, Robert E. Krieger, Malabar, Florida, 1965; Pharmaceutical Dosage Forms: Dispersed Systems Vol. I, Martin M. Rigear, US Pat. No. 5,595,723, assigned to Surfactants and Sonus Pharmaceuticals, the assignee of this invention. All these reference data are incorporated herein by reference.

TPGS : TPGS and PEGylated Vitamin E are vitamin E derivatives to which polyethylene glycol subunits are attached by succinic acid diesters in the cyclic hydroxyl of the vitamin E molecule. TPGS stands for D-α-tocopherol polyethylene glycol 1000 succinate (MW = 1513). TPGS is a nonionic surfactant (HLB = 16-18) having the structure of Formula III:

Various chemical derivatives of vitamin E TPGS, including ethers and ether bonds of various chemical moieties, fall within the scope of vitamin E TPGS.

Polyethylene glycol : Polyethylene glycol (PEG) is a hydrophilic polymerized form of ethylene glycol composed of repeating units of chemical structure-(CH ₂ -CH ₂ -O). The general formula for polyethylene glycol is HOCH ₂ (CH ₂ OCH ₂ ) _n CH ₂ —OH or H (OCH ₂ OCH ₂ ) _n OH. The molecular weight ranges from 200 to 10,000. These various forms are described as PEG-200, PEG-400 and the like.

N-methyl-2-pyrrolidone : N-methyl-2-pyrrolidone (NMP) is an organic molecule having the following chemical structure:

The GMP grade of this compound can be purchased under the name Pharmasolve ^{TM and} is used to improve the solubility of poorly soluble drugs in pharmaceutical compositions. The enhanced solubility of certain drugs can be attributed to the complexation of the molecule with the nitrogen and carbonyl reactive centers.

Polyvinyl Pyrrolidone : Polyvinyl pyrrolidone (PVP) or Povidone is a water soluble polymer and consists of repeating units of the following chemical structure:

Average MW can vary from 2500 to 3 × 10 ⁶ express pyrrogen-povidone, which can be used for parenteral administration. Concentrations up to 5% w / v can be used as cosolvents for poorly soluble drugs.

Poloxamer or Pluronic : This is a synthetic block copolymer of ethylene oxide and propylene oxide having the following general structure.

The following isomers, based on the values of a and b, can be purchased from BASF Performance Chemicals (Parsippany, New Jersey) under the trade name Pluronic, and under the CTFA designation Poloxamer 108, 188, 217, 237, 238, 288, 338 , 407, 101, 105, 122, 123, 124, 181, 182, 183, 184, 212, 231, 282, 331, 401, 402, 185, 215, 234, 235, 284, 333, 334, 335, and It consists of a group of surfactants called 403. The values of a and b for the most commonly used poloxamers 124, 188, 237, 338 and 407 are 12/20, 79/28, 64/37, 141/44 and 101/56, respectively.

Solutol HS-15 : This is polyethylene glycol 660 hydroxystearate from BASF (Parsippany, NJ). In addition to free polyethylene glycol and its monoesters, di-esters can also be detected. According to the manufacturer, a typical lot of Soluble HS-15 contains approximately 30% free polyethylene glycol and 70% polyethylene glycol esters.

Other Surfactants Other surfactants useful in the present invention include ascorbyl-6-palmitate (Roche Vitamines, Nutley NJ), stearylamines, and sucrose fatty acid esters (Mitsubishi Chemicals). Typical surfactants include compounds with polar hydrophilic heads and hydrophobic tails, such as vitamin E derivatives including peptide bound polyglutamate and pegylated phytosterols attached to ring hydroxyls. Other peptides can also be bound to vitamin E. PEGylated phospholipids are also useful surfactants. Examples of pegylated phospholipids are C ₆ -C _{24 in} which the fatty acrylic chain can be saturated and unsaturated. PEG 2000 or PEG 5000 analogs of phosphatidylethanolamine including fatty acids, mixtures thereof.

Hydrophilic-lipophilic balance : An empirical approach was used to index surfactants. Its value varies from 1 to 45, and about 1 to 20 for nonionic surfactants. In general, the HLB for lipophilic surfactants is less than 10 and the HLB for hydrophilic surfactants is greater than 10.

Biocompatibility : It is one that can function in an acceptable manner in or on a living organ, without unnecessary toxicity or physiological or pharmaceutical effects.

Substantially free monohydric alcohol : This is a composition having a monohydric alcohol at a monohydric alcohol concentration of less than about 1.0% (w / v). As used herein, the term “monovalent” alcohol is an alcohol containing one hydroxyl group, such as but not limited to ethanol, butanol, isopropanol. The term "polyhydric" alcohol or "polyol" is an alcohol containing two or more hydroxyl groups, such as but not limited to ethylene glycol, propylene glycol or polyethylene glycol (PEG). PEG is also a "polyglycol" having ethylene glycol as polymerized unit. Other suitable polyhydric alcohols for use herein include but are not limited to ethylene glycol (two-OH groups), glycerol (three-OH groups), sorbitol (six-OH groups) and mannitol (six-OH groups). It is not limited.

Emulsion : A colloidal dispersion of two immiscible liquids, typically in the form of optically clear droplets, typically between 0.1 and 0.3 microns in diameter and the dispersed continuous phase is not a matched refractive index. Such systems have finite stability, generally defined by the present application or related literature system, which can be enhanced by the addition of amphoteric molecules or viscosity enhancers.

Microemulsion : A thermodynamically stable isotropic transparent dispersion of two immiscible liquids, stabilized by an interfacial film of surfactant molecules, such as oil and water. Microemulsions have an average droplet diameter of less than 200 nm, usually between 10 and 50 nm. In the absence of water, the mixture of oil and nonionic surfactant forms a clear and isotropic solution known as a self-emulsifying drug delivery system (SEDDS) and is successful in improving lipophilic drug dissolution and oral absorption. Has been used.

PEGylated : PEGylated or ethoxylated refers to polyethylene glycol subunits attached to a given compound via chemical bonds.

Aqueous media : pharmaceutically acceptable such as acidifying agents, alkalizing agents, buffering agents, chelating agents, complexing and solubilizing agents, antioxidants and antimicrobial preservatives, moisturizing suspensions and / or viscosity modifiers, osmotic and wettable or other biocompatible materials. Water-containing liquids which may include possible additives.

Therapeutic : This is a synthetic natural compound with biological activity that has at least two octanol-buffer partition coefficients (Log P) to ensure that it can be dissolved in the oil phase and the therapeutic agent preferentially dissolves in the oil phase rather than in the water phase. . This includes peptides, non-peptides and nucleotides. Hydrophobic derivatives of water soluble molecules such as fatty acids and lipid conjugates / prodrugs are within the scope of therapeutic agents.

Chemotherapeutic agents : These are natural or synthetic molecules that are effective against one or more cancerous forms, in particular molecules that may be slightly or completely lipophilic or may be changed to lipophilic. This definition includes molecules that are cytotoxic substances (anticancer agents), those that stimulate the immune system (immunostimulators) and modulators of angiogenesis by their mechanism of action. The results in either case slow the growth of cancer cells.

Chemotherapeutic agents include paclitaxel and related molecules called taxoids, taxins or taxanes by collective nouns. The structure of paclitaxel is shown in the following figure (Formula VI).

The definition of "taxoid" includes attachment to a basic ring structure (taxoid nucleus), which is effective in reducing various cell changes and cancer cell growth and may indicate cleavage into oil (fatty phase), which is known to those skilled in the art. It can be configured by organic chemical techniques known to them. These are benzoate derivatives of paclitaxel such as 2-debenzoyl-2-aroyl and C-2-acetoxy-C-4-benzoate paclitaxel, 7-deoxytaxol, C-4 aziridine paclitaxel, in particular BMS-188797 Methyl carbonate derivatives of paclitaxel, as well as natural and synthetic polymers, in particular fatty acids, phospholipids, and various paclitaxel conjugates of glycerides with 1.2-diasiloxypropan-3-amine. The structure of the taxoid nucleus is shown in formula (VII).

Within the scope of the present invention are included natural products that share structural similarities with paclitaxel, ie they include conventional drug delivery agents proposed for microtubule-stabilizers. These compounds include, but are not limited to, epothilones A and B, discodermolide, nonataxel and eleuterobin (Chem. Eng. News 1999, 77 (17): 33-36).

Chemotherapeutic agents include podophyllotoxin and derivatives and analogs thereof. The core ring structure of these molecules is shown in the following figure (Formula VIII):                 

Another important class of chemotherapeutic agents useful in the present invention is campotesine, whose common ring structure is shown in the following figure, which retains the efficacy of the following molecule (Formula IX) and preserves its lipophilic properties. Variations on are also included.

Another preferred class of chemotherapeutic agents useful in the present invention is lipophilic anthracycline, the basic ring structure of which is shown in the following figure (Formula X):                 

Suitable lipophilic modifications of formula (X) include substitutions at ring hydroxyl groups or sugaramino groups.

Another important class of chemotherapeutic agents may be lipophilic or lipophilic by molecular chemical synthesis alterations well known to those skilled in the art, for example by combinatorial chemistry and molecular modeling, Derived: Taxotere, Amonafide, Illudin S, 6-hydroxymethylacylpulben Bryostatin 1,26-succinylbriostatin 1, Palmitoyl Rhizoxin, DUP 941, Mitomycin B, Mitomycin C, Penclomedine. Interferon α2b, angiogenesis inhibitor compounds, hydrophobic complexes such as 2-hydrazino-4.5-dihydro-1H-imidazole with platinum chloride and 5-hydrazino-3,4-dihydro-2H-pyrrole with platinum chloride , Vitamin A, vitamin E and derivatives thereof, especially tocopherol succinate.

Other compounds useful in the present invention include: 1,3-bis (2-chloroethyl) -1-nitrosurea ("carmustine" or "BCNU"), 5-fluorouracil, doxorubicin (" Adriamycin "), epirubicin, aclarubicin, Bisantrene (bis (2-imidazolen-2-ylhydrazone) 9,10-anthracenedicarboxaldehyde, mitoxantrone, methotrexate, etatrexate , Muramil tripeptide, Muramil dipeptide, lipopolysaccharide, 9-bd-arabinofuranosyladenine ("Vidarabine") and 2-fluoro derivatives thereof, resveratrol, retinoic acid and retinol, Cartenids, tamoxifen .

Other compounds useful in the application of the present invention include: Decarbazine, Lonidamine, Proxantrone, Anthrapyrazoles, Etoposide, Camptothecine, 9-aminocamptothecin, 9-nitrocamptothecin, camptothecin-11 ("Irinotecan" ), Topotecan, Bleomycin, Vinca alkaloids and analogs thereof [Vincristine, Vinorelbine, Vindesine, Vintripol, Vinxaltine, Ancitabine], 6-aminocrisene, and navelvin.

Other compounds useful in the application of the present invention are the mimetics of taxol, eleuterobin, sacroditin, discodermolide and epotiolone. Other compounds useful in the present invention are microtubule targeting agents. Microtubule targeting agents bind to a protein called tubulin to prevent microtubule polymerization. Representative microtubule binders include epothilones, eruterobin and discodermolide.

Valproic acid, tacrolimus, rapamycin, clarithromycin, erythromycin, neomycin, vacitracin, tyrotropin, somatostatin, testosterone, progesterone, cortisone, polyketide class, quinolone class, ciprofloxacin, benzodiazepine class Diazepam, calcitriol, clozapine, androcepin are also useful.

In view of these definitions, the present invention relates to pharmaceutical compositions in the form of self-emulsifying drug delivery systems free of emulsions, micelle solutions or substantially ethanol solvents.

The therapeutic agent of the composition of the present invention is initially dissolved in the cosolvent. When ethanol is used as the treating solvent during the preparation of the oil phase, the ethanol is removed and a substantially ethanol free composition is formed. The ethanol concentration is 1% (w / v), preferably less than 0.5%, most preferably less than 0.3%. The therapeutic agent may also be dissolved in methanol, propanol, chloroform, isopropanol, butanol and pentanol. These solvents are also removed before use.

In a preferred embodiment, the therapeutic composition of the invention is benzyl alcohol, benzyl benzoate, dimethyl sulfoxide (DMSO), dimethylamide (DMA), propylene glycol (PG), polyethylene glycol (PEG), N-methyl-2-pi It can be dissolved in nonvolatile cosolvents such as rolidone (NMP) and polyvinylpyrrolidone (PVP). Particular preference is given to water soluble polymers such as NMP or PEG or PVP (Table 1).

The main advantage / improvement of using PEG-400 over alcohols such as ethanol to dissolve the therapeutic agent is that there is no need to remove or dilute the volatile solvent prior to administering the therapeutic agent. The final polyethylene glycol level in the emulsion may vary from 1 to 50%, preferably 1 to 25% more preferably 1 to 10% (w / w). Suitable polyethylene glycol solvents are those having an average molecular weight of 200 to 600, preferably 300 to 400 (Table 1). In the case of self-emulsifying systems for oral administration, high molecular weight PEG (1,000-10,000) can also be included as a coagulant to form a semisolid formulation that can be filled into a rigid gelatin capsule.

Physical Properties of Low Molecular Weight Polyethylene Glycol Physical properties PEG 200 PEG 300 PEG 400 PEG 600 Molecular Weight 190-210 285-315 380-420 570-630 Viscosity (mPas) 46-53 66-74 85-95 130-150 Refractive Index (25 ℃) 1.459 1.463 1.465 1.467 Freezing point (℃) -50 -16 to -12 -3 to 8 15 to 25

Solubilization of the therapeutic agents of the present invention in polyethylene glycol or other non-volatile cosolvents (PVP, NMP) eliminates the need for solubilizing the therapeutic agents of the invention in monohydric alcohols such as ethanol or other volatile solvents. Polyethylene glycol or N-methyl-2-pyrrolidone eliminates the need to remove the solvent prior to use of the therapeutic emulsion.

The final polyethylene glycol level in the emulsion can vary from 1 to 50%, preferably from 1 to 25% more preferably from 1 to 10% (w / w).

The composition of the present invention contains tocopherol (preferably α-tocopherol) as a carrier for therapeutic drugs, and can be administered to animals or humans through intravascular, oral, intramuscular, skin and subcutaneous routes. Specifically, emulsions can be provided by the following routes: intraperitoneal, intraarterial, intraarticular, intrafocal, intracranial, intracranial, intraluminal, intradural, intralesional, intraocular, lobe, intrawall, intraocular, intraoperative , Intramural, intraperitoneal, pleural, intrapulmonary, intramedullary, intrathoracic, intramedural, tympanic, intrauterine, and intraventricular percutaneous. Emulsions of the present invention can be sprayed using suitable aerosol propulsion devices known in the art for the delivery of lipophilic compounds.

In a first aspect, the present invention relates to the use of tocopherol as a hydrophobic disperse phase of an emulsion containing a water insoluble, poorly water soluble therapeutic agent, a lower water soluble modified therapeutic agent or a mixture thereof. In a preferred embodiment α-tocopherol is used. Α-tocopherol, also called "vitamin E", is not a typical lipid oil. It exhibits higher polarity than most lipid oils, especially triglycerides and cannot be saponified. It is practically insoluble in water.

In a second aspect, the invention is a tocopherol emulsion in the form of a self-emulsifying system, which system can be used for oral administration of a water insoluble (or poorly water soluble or water soluble agent or mixture thereof) if desired. . In this embodiment, the oil phase with the surfactant and the drug or drug mixture may be wrapped in a soft or hard gelatin capsule. Suitable coagulants with a melting point of 40 to 60 ° C., such as high molecular weight polyethylene glycol (MW> 1000) and glycerides such as those sold under the trade name Gelucire (Gattefose Corp, Saint Priest, France), are added to hard gelatin at high temperatures. The capsule can be filled with the preparation. Semisolid preparations are formed at room temperature equilibrium. During dissolution of gelatin in the stomach and duodenum, the oil is released and naturally forms a fine emulsion having an average droplet diameter of 2 to 5 microns. The emulsion is then ingested by the intestinal microvilli and released into the bloodstream.

In a third aspect, the invention comprises a microemulsion containing at least one tocol, preferably α-tocopherol. Microemulsions refer to a subclass of emulsions that are infinitely stable thanks to the microaggregates of oil / drugs of extremely small size in which the emulsion suspension is essentially transparent and dispersed therein.

In a fourth aspect of the invention, PEGylated vitamin E (TPGS) is used as the primary surfactant in vitamin E emulsions. PEGylated vitamin E is used as primary surfactants, stabilizers and also as co-solvents in vitamin E emulsions. Polyethylene glycol (PEG) is also useful as a cosolvent in the emulsion of the present invention. In particular polyethylene glycol 200, 300, 400 or mixtures thereof are used.

The concentration of α-tocopherol and / or other tocol in the emulsion of the invention may be about 1 to about 20% w / w. When TPGS is present in the composition, the ratio of tocopherol to TPGS is optionally from about 1: 1 to about 20: 1.

The emulsions of the invention are nonionic, such as ascorbyl-6-palmitate, stearylamine, pegylated phospholipids, sucrose fatty acid esters, various tocol derivatives and polyoxyptopylene-polyoxyethylene glycol nonionic copolymers, Synthetic surfactant mixture.

The emulsion of the present invention may comprise an aqueous medium. The aqueous phase has an osmolality of approximately 300 mOsm and may include sodium chloride, sorbitol, mannitol, polyethylene glycol, propylene glycol albumin, polypeptides and mixtures thereof. This medium may contain various additives to help stabilize the emulsion or impart biocompatibility to the composition. Acceptable additives may include acidifying agents, alkalizing agents, antimicrobial preservatives, antioxidants, buffering agents, chelating agents, suspending and / or viscosity-increasing agents, and osmotic agents. Preferably, agents that control pH, osmoticity and increase viscosity are included. Optionally, at least 250 mOsm is obtained using agents such as sorbitol or sucrose, which also increase the viscosity.

The emulsion of the invention for intravenous injection has a particle size (average droplet diameter) of 10 to 500 nm, preferably 10 to 200 nm and most preferably 10 to 100 nm. For intravenous emulsions, the spleen and liver will remove particles larger than 500 nm.                 

Preferred forms of the present invention comprise cytotoxin, which is very insoluble in water used for the treatment of paclitaxel, ie uterine cancer and other carcinomas. The emulsion composition of the present invention comprises vitamin E containing paclitaxel at a concentration of up to 20 mg / mL, four times what is currently available as a prescription drug, so that the emulsion microdroplets are less than 0.2 micron and can be finally sterilized by filtration, And a solution consisting of a biocompatible surfactant.

Preferred injectable compositions include: 0.1-1.0% paclitaxel (1-10 mg / ml); 1 to 10% PEG 400; 1 to 20% vitamin E; 1-10% TPGS and 0.5-2.5% Poloxamer 407. Another preferred composition includes: 1.0% paclitaxel (10 mg / ml), 6% PEG 400, 8% Vitamin E, 5% TPGS, 1% Pluronic F127 and 80% aqueous solution.

Preferred preparations for the self-emulsification system are as follows: 0.1-20% paclitaxel, 10-90% vitamin E, 10-90% PEG 400 or N-methyl-2-pyrrolidone, 5-50% TPGS, 5 to 50% secondary hydrophilic surfactants such as Polysorbates (Tween 80), Pluronics (Pluronic F127) or Cremophor EL / RH40, Solutol HS-15. The oily phase (vitamin E) optionally contains polyvinylpyrrolidone, glycerol and propylene glycol esters such as mono- / di- / triglycerides and mono-diesters. Moreover, high molecular weight PEG (1,000 to 10,000 MW) and high melting point glycerol esters can be included to provide preparations with semisolid persistence.

Another embodiment of the present invention includes carcinomas comprising parenteral administration of paclitaxel feedback in vitamin E emulsions with or without PEGylated vitamin E by intravenous injection once or twice a day over several weeks of treatment. Is a treatment method. Such methods can be used to treat carcinoma of the breast, lung, skin and uterus.

The invention will be better understood with reference to the following drawings:

1A shows the particle size of paclitaxel emulsion (QWA) over time at 7 ° C .;

1B shows the particle size of paclitaxel emulsion (QWA) over time at 25 ° C .;

2 is an HPLC chromatogram showing the integrity of paclitaxel in an emulsion as described in Example 5;

3A shows paclitaxel concentration of paclitaxel emulsion (QWA) over time at 4 ° C .;

3B shows paclitaxel concentration of paclitaxel emulsion (QWA) over time at 25 ° C .;

Figure 4 shows the percentage of paclitaxel released from three different emulsions over time. The mark • indicates the percentage of paclitaxel released over time from an emulsion commercially available from Bristol Myers Squibb. Mark ▲ indicates the percentage of paclitaxel released over time from the emulsion of the invention (QWA) containing 6 mg / ml paclitaxel (QWA) as shown in Example 6. Indication o indicates the percentage of paclitaxel released over time from the emulsion of the invention (QWB) containing 7 mg / ml paclitaxel as shown in Example 7.

5 shows the efficacy of PEG-400 / vitamin E / paclitaxel emulsions on B16 melanoma in mice.

Alternate principles of the invention may be more fully understood by reference to the following non-limiting examples.

Example 1

Dissolution of Paclitaxel in α-tocopherol

α-tocopherol was purchased from Sigma Chemical Company (St Louis MO) in the form of 95% purity synthetic di-α-tocopherol made from phytol. This oil was amber and very viscous. Paclitaxel was purchased from Hauser Chemical Research (Boulder CO), which was 99.9% pure by HPLC. 200 mg of paclitaxel was dissolved in 6 mL dry anhydrous ethanol (Spectrum Chemical Manufacturing Corp. Gardena CA) and added to 1 gm of α-tocopherol. The ethanol was then removed in vacuo at 42 ° C. until the residue had a constant mass. Independent studies showed that the ethanol content was less than 0.3% (w / v).

The resulting solution was clear, amber and very viscous and had a nominal concentration of 200 mg / gm (w / w) in α-tocopherol. Higher paclitaxel concentrations (400 mg / gm, w / w) can be dissolved in α-tocopherol.

Example 2

Anionic Surfactants Used to Prepare α-Tocopherol Emulsions

2 gm of paclitaxel in 10 gm α-tocopherol prepared as described in Example 1 was emulsified using ascorbyl palmitate as the triethanolamine salt by the following method. A solution consisting of 20 mM ascorbic acid was buffered to pH 6.8 using triethanolamine as the free base to form a 2x buffer. 50 mL of 2x buffer was placed in the Waring blender. Anionic surfactant 0.5 gm of ascorbyl-6-palmitate (Roche Vitamins and Fine Chemicals, Nutley NJ) was added and the solution was mixed for 2 minutes at high temperature at 40 ° C. under argon. Α-tocopherol containing paclitaxel was then added to the mixer along with the surfactant and buffer. Mixing was continued until a milky pre-emulsion of the crude particles was obtained after approximately 1 minute at 40 ° C. Water for injection was then added to bring the final volume to 100 ml.

The preemulsion was delivered to feeder Microfluidizer Model 110Y (MicroFluidics Inc, Newton MA). During homogenization, the unit was immersed in a bath to maintain a process temperature of approximately 60 ° C. and washed with argon before use. After preparation, the emulsion was recycled through a homogenizer continuously for 10 minutes at a pressure gradient of about 18 kpsi across the interaction head. The flow rate was about 300 mL / min, which in turn indicates about 25 passes of the homogenizer.

The resulting paclitaxel emulsion in the α-tocopherol excipient was placed in amber vials under argon and refrigerated at 7 ° C and 25 ° C. Samples were taken at separate time periods for particle size and chemical analysis.                 

Data obtained with a Nicomp Model 370 Submicron Particle Sizer (Particle Sizing Systems Inc. Santa Barbara CA) showed that the emulsion had an average particle diameter of 280 nm.

Example 3

Use of PEGylated Vitamin E (TPGS)

Three-component state diagrams were prepared for α-tocopherol, PEGylated Vitamin E (TPGS, Vitamin-E polyoxyethylene glycol-1000-succinate, purchased from Eastman Chemical Co., Kingsport TN), and water. TPGS was first melted at 42 ° C. and mixed by weight with α-tocopherol in various portions of 1-100% TPGS, the balance being α-tocopherol. The mixture was miscible in all compositions. Water was then added to each mixture in such a way that the final water concentration increased from 0 to 97.5% in steps. In each step, the phase behavior of the mixture was observed. Suitably, the mixture was mixed by vortex and sonication and the mixture was heated or centrifuged to estimate its compatibility.

Large areas of biphasic o / w emulsions suitable for parenteral administration have been found at water concentrations above 80%. The emulsion formed was a milky white free flowing liquid and contained dispersed α-tocopherol microparticles stabilized by a nonionic surfactant. Also in this area, microemulsions that could be suitable as drug carriers were observed at a ratio of TPGS to oil of greater than about 1: 1. At low water contents a large area containing a clear gel (reverse emulsion) appeared. The separation of the two regions (high water content and low water content) is an opaque soapy liquid crystal.                 

TPGS or drugs with α-tocopherol and surfactant combinations, eg, nonionic, anionic or cationic co-surfactants (eg glutamyl stearate, ascorbyl palmitate or Pluronic F-68) The phase diagram of can be prepared in a similar manner.

Example 4

Α-tocopherol emulsion for intravenous delivery of paclitaxel

Preparations of the following compositions were prepared:

Paclitaxel 1.0 gm%

α-tocopherol 3.0 gm%

TPGS 2.0 gm%

Ascorbyl-6-palmitate 0.25 gm%

Sorbitol 5.0 gm%

To triethanolamine pH6.8

With 100 mL qs of water

The preparation method is as follows: Synthetic α-tocopherol (Roche Vitamins, Nutley NJ), paclitaxel (Hauser, Boulder CO), ascorbyl 6-palmitate (Aldrich Chemical Co, Milwaukee WI) and TPGS at 40-45 ° C. It was dissolved in 10 volumes of anhydrous unmodified ethanol (Spectrum Quality Products, Guardenia CA) while heating. The ethanol was then evacuated using vacuum until it remained below 0.3 wt%.

The pre-warmed aqueous solution containing the biocompatible osmotic material and buffer was added with moderate mixing to form a white milky immediately. This mixture can be further improved with moderate rotation for 10 minutes while continuously warming to 40-45 ° C. This premix was then further emulsified at pH 7 as follows.

The premix at 40-45 ° C. was homogenized in an Avestine C5 homogenizer (Avestin, Ottawa Canada) at 26 Kpsi at 44 ° C. for 12 minutes. The resulting mixture contained α-tocopherol microparticles having an average particle size of about 200 nm. The pH was further adjusted using 1 M triethanolamine (Spectrum Quality Products) alkaline solution.

To avoid gelation of the TPGS during the initial emulsification, all operations were performed at temperatures above 40 ° C. and care was taken not to expose the solution to cold air by covering all containers containing the mixture. Secondly, less than 2% of TPGS should generally be dissolved in α-tocopherol oil prior to preemulsification, and the balance of TPGS is first dissolved in the aqueous buffer before the preemulsion is prepared. The solution gels at a concentration of TPGS> 2%.

The physical stability of the emulsion was then observed by storing a number of vials at 4 ° C and 25 ° C. Over several months, the vials were periodically taken out and particle size was measured. Average particle sizes are shown for two storage temperatures in FIG. 1 as measured by Nicomp Model 370 (Particle Sizing Systems Inc. Santa Barbara CA). The particle size distribution was two modes.

Example 5

error! Undefined bookmark, chemical stability of paclitaxel in α-tocopherol emulsion

After emulsification, the preparation of Example 4 was analyzed for paclitaxel on a Phenosphere CN column (5 microns, 150 × 4.6 mm). The fluidized bed consisted of a methanol / water gradient and had a flow rate of 1.0 mL / min. Paclitaxel was detected and measured using a UV detector set at 230 nm. A single peak was detected (FIG. 2), which had a retention time and mass spectrum picture consistent with the native reference paclitaxel purchased from Hauser Chemical (Boulder CO).

Example 4 The chemical stability of the emulsion was measured by HPLC during storage. The data in FIG. 3 show that paclitaxel remains stable in the emulsion for a period of at least 3 months regardless of storage temperature. In addition, Figures 2 and 3 show the successful maintenance and emulsion stability of drug efficacy when stored for a period of at least 3 months at 4 ° C.

Example 6

Paclitaxel Emulsion Formulation QWA

A 10 mg / ml paclitaxel emulsion for intravenous drug delivery, having the following composition, was prepared as described in Example 4.

Paclitaxel 1.0 gm%

α-tocopherol 3.0 gm%

TPGS 1.5 gm%

Ascorbyl-6-palmitate 0.25 gm%                 

Sorbitol 4.0 gm%

To triethanolamine pH6.8

With 100 mL qs of water

Example 7

Paclitaxel Emulsion Formulation QWB

A second emulsion of 10 mg / ml paclitaxel for intravenous drug delivery, with the following composition, was prepared as described in Example 4.

Paclitaxel 1.0 gm%

α-tocopherol 3.0 gm%

TPGS 1.5 gm%

Solutol HS-15 1.0 gm%

Sorbitol 4.0 gm%

To triethanolamine pH6.8

With 100 mL qs of water

Solutol HS-15 is a product of BASF Corp. (Mount Olive NJ).

Example 8

10 mg / mL paclitaxel emulsion preparation QWC

A third emulsion of 10 mg / ml paclitaxel was prepared as follows using Poloxamer (BASF Corp, Parsipany NJ) as co-surfactant.

Paclitaxel 1.0 gm%                 

α-tocopherol 6.0 gm%

TPGS 3.0 gm%

Poloxamer 407 1.0 gm%

Sorbitol 4.0 gm%

To triethanolamine pH6.8

100 mL qs of water for injection

In this example, 1.0 mg Poloxamer 407 and 1.0 mg paclitaxel were dissolved in moderately heated 6.0 gm α-tocopherol with 10 volumes of ethanol. Ethanol was then removed under vacuum. Independently, an aqueous buffer was prepared by dissolving 3.0 gm TPGS and 4.0 gm sorbitol in 90 mL of final volume of water for injection. Both oil and water solutions were warmed to 45 ° C. and mixed by sonication to prepare a preemulsion. Vacuum was used to remove excess air from the preemulsion before homogenization.

Homogenization was performed on Avestine C5, already described. The pressure difference across the homogenization valve was 25 Kpsi and the feed temperature was 42-45 ° C. A chiller was used so that the product from the homogenizer did not exceed a temperature of 50 ° C. A flow rate of 50 mL / min was obtained during homogenization. After about 20 passes in the recycle mode, the emulsion became more cloudy. A sample was taken and sealed in a vial as above. A fine α-tocopherol emulsion for intravenous delivery of paclitaxel was obtained. The average particle diameter of the emulsion was 77 nm. After sterile filtration through 0.22 micron Durapore filter (Millipore Corp, Bedford MA), the emulsion was filled into vials and stored at 4 ° C. until used for intravenous injection.

Example 9

5 mg / mL paclitaxel emulsion preparation QWC

Additional paclitaxel emulsions were prepared as described in Example 8 except that 5 mg / ml was incorporated instead of 10 mg / ml of the drug. The composition of the emulsion is as follows:

Paclitaxel 0.5 gm%

α-tocopherol 6.0 gm%

TPGS 3.0 gm%

Poloxamer 407 1.0 gm%

Sorbitol 4.0 gm%

To triethanolamine pH6.8

100 mL qs of water for injection

After homogenization as described in Example 8, a somewhat cloudy emulsion of α-tocopherol and paclitaxel having an average particle diameter of 52 nm was obtained. After sterile filtration through 0.22 micron Durapore filter (Millipore Corp, Bedford MA), the emulsion was filled into vials and stored at 4 ° C. until used for intravenous injection. Drug loss during filtration was less than 1%.

Example 10

Paclitaxel Emulsion Formulation QWD

A fifth emulsion of α-tocopherol for intravenous administration of paclitaxel was prepared as follows:

Paclitaxel 0.5 gm%

α-tocopherol 6.0 gm%

TPGS 3.0 gm%

Poloxamer 407 1.5 gm%

Sorbitol 4.0 gm%

To triethanolamine pH6.8

100 mL qs of water for injection

Synthetic α-tocopherol USP-FCC purchased from Roche Vitamines (Nutley, NJ) was used in this formulation. Polyethylene glycol 200 (PEG-200) was purchased from Sigma Chemical Co.

After homogenization, a slightly cloudy emulsion with an average particle diameter of 60 nm was obtained. After sterile filtration through 0.22 micron Durapore filter (Millipore Corp, Bedford MA), the emulsion was filled into vials and stored at 4 ° C. until used for intravenous injection. Drug loss during filtration was less than 1%.

Example 11

Dissolution of Paclitaxel in TGPS and Preparation of Micelle Solution

The inventors observed good solubility of paclitaxel in TGPS, about 100 mg drug per 1.0 TGPS. A micelle solution of TGPS containing paclitaxel was prepared as follows. Stock solution of paclitaxel in TGPS was prepared by dissolving 90 mg paclitaxel in 1.0 gm TPGS at 45 ° C. with ethanol to be removed under vacuum. A series of dilutions were then prepared by diluting the paclitaxel stock with additional TGPS to obtain paclitaxel in TPGS at concentrations of 0.1, 1, 5, 10, 25, 50, 75 and 90 mg / mL. A new test tube was used to dissolve each 100 mg of paclitaxel concentrate in TPGS in 0.9 mL water. All test tubes were mixed by vortexing and sonication at 45 ° C. Transparent micelle solutions in water corresponding to final paclitaxel concentrations of 0.01, 0.1, 0.5, 1.0, 2.5, 5.0, 7.5 and 9.0 mg / mL were obtained.

The solution was investigated using a Nicomp Model 370 laser particle sizer (Particle Sizing Systems Inc. Santa Barbara CA). A particle size of approximately 10 nm was obtained, which is consistent with the presence of micelles of TPGS and paclitaxel.

Paclitaxel micelle solution in TPGS containing up to 2.5 mg / mL paclitaxel was stable for at least 24 hours while unstable at 5.0, 7.5 and 9.0 mg / mL and drug crystals were rapidly irreversibly produced. These observations suggest that paclitaxel remains dissolved only in the presence of α-tocopherol-rich domains in the emulsion particles. Thus, an optimal ratio of α-tocopherol to TPGS is required to produce an emulsion in which high concentrations of paclitaxel can be stabilized.

When adjusted to appropriate osmoticity and pH, micelle solutions are effective for slow IV instillation of paclitaxel in cancer patients although AUC is expected to be low.

The effectiveness of TPGS in α-tocopherol is a synergy of several desirable properties. First, it has its own affinity for paclitaxel, possibly thanks to α-tocopherol, which forms a hydrophobic part of its molecular structure. Secondly, the interfacial tension between TPGS and α-tocopherol in water is about 10 dynes / cm, and this tension is sufficient to emulsify free α-tocopherol, especially when used with co-surfactants. Third, polyoxyethylated surfactants, such as TPGS, have excellent properties that are well formed as a "secret film" for injectable particles by dramatically reducing the trap of particles in the liver and spleen, as is well known in the art. However, the unexpected and unique discovery of TPGS as a surfactant for α-tocopherol emulsions was the discovery of all three properties in a single molecule. An additional advantage of TPGS is the fact that it forms a stable self-emulsifying system in oils and mixtures with solvents such as propylene glycol and polyethylene glycol, suggesting synergy when used with α-tocopherol for oral drug delivery. .

When adjusted to appropriate osmoticity and pH, micelle solutions are effective for slow IV instillation of paclitaxel in cancer patients although AUC is expected to be low.

Example 12

20 mg / mL paclitaxel emulsion preparation

A crude particle emulsion containing 20 mg / mL paclitaxel in α-tocopherol was obtained by the method described in Example 4 using 5% α-tocopherol and 5% TPGS, ie simply by increasing the concentration. Efforts to test higher concentrations have not been made because no further increase in concentration is needed for clinically useful intravenous emulsions.

Example 13

Use of Other PEG Surfactants in α-Tocopherol Emulsions

Various other PEGylated surfactants such as Trioton X-100, PEG 25 propylene glycol stearate, Brij 35 (Sigma Chemical Co), Myrj 45, 52 and 100, Tween 80 (Spectrum Quality Products), PEG 25 Glyceryl Triolade (Goldschmit Chemical Corp. Hopewell VA) is effective for emulsifying α-tocopherol.

However, experiments with several other pegylated surfactants failed completely to stabilize paclitaxel in the α-tocopherol emulsion. Three emulsions were prepared as described in Example 9 except for replacing TPGS with Tween 80 and Myrj 52 as primary surfactants in each emulsion to demonstrate the unique effectiveness of TPGS. These two surfactants were chosen because Tween 80 and Myrj 52 had essentially the same HLB value as TPGS, resulting in a fairly good α-tocopherol emulsion. However, when 5 mg / mL paclitaxel is included in the preparation, crystallization of the drug appears very rapidly after preparation of the pre-emulsion, and treated emulsions of Tween 80 and Myrj 52 are rod-shaped particles up to several microns in length. It is characterized by a co-particle matching the paclitaxel crystals. Unlike TPGS emulsions, which easily pass a 0.22 micron filter with less than 1% drug loss, Tween and Myrj emulsions could not be filtered due to the presence of this crystalline drug material.

There are several possible explanations for the unexpected improvement of α-tocopherol paclitaxel emulsions with TPGS. The drug has good solubility up to about 100 mg / mL in TPGS. Perhaps it is the affinity strength of the paclitaxel benzyl side chain with the planar α-tocopherol phenol ring in the TPGS molecule which stabilizes the complex of drug and carrier. In addition to the succinate linkers of α-tocopherol and PEG tails, a novel feature of this molecule is that its structure differs from other PEGylated surfactants tested.

Example 14

Poloxamer-Based α-Tocopherol Emulsion

α-tocopherol 6.0 gm%

Poloxamer 407 2.5 gm%

Ascorbyl palmitate 0.3 gm%

Sorbitol 6.0 gm%

Triethanolamine to pH7.4

With 100 mL qs of water

α-tocopherol emulsion was prepared using Poloxamer 407 (BASF) as the primary surfactant. The white milky evi mixture was homogenized by continuous recirculation at 25 Kpsi for 10 min in a C5 homogenizer (Avestin, Ottawa Canada) with a feed temperature of 45 ° C. and chiller outlet for the product set to 15 ° C. A fine, sterile, soluble emulsion of α-tocopherol microparticles was obtained. However, when this preparation is prepared using paclitaxel, precipitation of paclitaxel appears after overnight storage in the refrigerator, which in turn lags in the good effectiveness of TPGS as the main surfactant.

Example 15

Lyophilized Emulsion Formulation

Maltrin M100 (Grain Processing Corporation, Muscatine IA) was added to the emulsion of Example 14 as a 2x stock in water. The aliquots were then frozen in a shell freezer and lyophilized under vacuum. A fine emulsion was recovered during dissolution in water.

Lyophilized preparations are effective where the indefinite shelf life of the lyophilized formulation is desired. Lyophilizable formulations containing other sugars such as mannitol, albumin or PolyPep from Sigma Chemicals, St. Louis, Mo. may also be prepared.

Example 16

In vitro release of paclitaxel from α-tocopherol emulsion

, Bristol Myers Squibb(BMS), Princeton NJ]에 비해 파클리탁셀의 지속적인 방출을 제공한다는 것을 발견하였다. 6mg/mL(QWA) 및 7mg/mL(QWB)의 파클리탁셀 농도를 가지는 에멀젼을 제조하였다. 비교로서, Taxol

은 에탄올:크레모포어 EL 1:1(v/v)에 용해된 6mg/mL의 파클리탁셀을 함유한다. 다른 조제물로부터 37℃에서 인산염-버퍼링된 식염수용액(PBS)으로의 체외 방출을 파클리탁셀에 자유롭게 투과할 수 있는 투석 멤브레인(MW 10킬로달톤의 컷-오프)을 사용하여 감시하였다. 투석 전과 후의 샘플에서 약물의 정량화를 HPLC로 행하였다. 약물의 방출형상을 퍼센트방출 및 시간에 따라 방출된 파클리탁셀의 농도 모두의 항으로 생성하였다. 도 4에서의 자료로부터 알 수 있는 바와 같이, 5% 미만의 파클리탁셀이 에멀젼으로 부터 24시간에 걸쳐 투석되었고, 한편 약 12%가 상용의 BMS 조제물로부터 투석 백의 외부에서 회수되었다. 이것은 에멀젼으로부터의 약물방출이 상업적으로 구입할 수 있는 용액에 비해 상당히 느렸다는 것을 나타낸다.One desirable property of drug delivery excipients is to provide sustained release of the incorporated drug, which is very often associated with improved pharmacokinetics and efficacy. In particular, long-circulating paclitaxel emulsions can improve the delivery of drugs to cancer sites in the body. The inventors surprisingly found that the emulsion of the present invention is currently the only FDA approved paclitaxel preparation [Taxol].

, Bristol Myers Squibb (BMS), Princeton NJ], have been found to provide sustained release of paclitaxel. Emulsions were prepared with paclitaxel concentrations of 6 mg / mL (QWA) and 7 mg / mL (QWB). As a comparison, Taxol

Silver contains 6 mg / mL paclitaxel dissolved in ethanol: cremophor EL 1: 1 (v / v). In vitro release from other preparations into phosphate-buffered saline solution (PBS) at 37 ° C. was monitored using a dialysis membrane (cut-off of MW 10 kilodaltons) freely permeable to paclitaxel. Quantification of the drug was performed by HPLC on samples before and after dialysis. The release form of the drug was generated in terms of both the percent release and the concentration of paclitaxel released over time. As can be seen from the data in FIG. 4, less than 5% of paclitaxel was dialyzed over 24 hours from the emulsion, while about 12% was recovered outside of the dialysis bag from a commercial BMS preparation. This indicates that drug release from the emulsion was significantly slower than commercially available solutions.

Example 17

Biocompatibility of α-Tocopherol Emulsions Containing Paclitaxel

Serious single-dose toxicity studies were performed. Each of 20-25 gm of mice was purchased and allowed to acclimatize in an approved animal facility. A group of mice (n = 3) was administered a formulation containing 30 to 90 mg / kg in α-tocopherol emulsion prepared as described in Example 6. All infusions were intravenously by tail vein reconstruction.

Although all infusions were done with a bolus IV push, no death or immediate toxicity was observed at any dose, even at 90 mg / kg. The results for body weight are shown in Table 2. The largest weight loss in the crowd was 17%, but in all groups, even 90 mg / kg, body weight recovered or gained over a period of 10 days post-infusion.

Excipient toxicity studies were also conducted. Animals given drug-free emulsions grew rapidly and gained weight slightly compared to animals administered with or without saline. This was due to the vitamin and calorie content of the preparation.

조제물은 10 mg/kg의 거환 정맥내 복용의 마우스에서 사망을 유발하였으며, 이 발견은 발명자들의 방법에서 반복되었다. 래트에서, Taxol

은 실험된 모든 희석액 및 복용영역에서 치명성이 일정하지 않았다. 역으로, 실시예 6의 조성물은 래트에서 잘 수용되었고, Rhone-Poulenc Rorer에 의해 상용화된 독성이 덜한 파클리탁셀 유사체인 심지어 Taxotere보다 개선된다. The inventors observed the highest acceptable dose (MTD) for paclitaxel over 90 mg / kg with no adverse effects. This was more than twice the best reported value of death observed at much lower doses. FDA-Approved Taxol

The preparation caused death in mice receiving a 10 mg / kg bolus intravenous dose, and this finding was repeated in our method. In rats, Taxol

The mortality was not constant in all dilutions and doses tested. Conversely, the composition of Example 6 was well received in rats and improved over even Taxotere, a less virulent paclitaxel analog that was commercialized by Rhone-Poulenc Rorer.

One possible explanation for high drug resistance is that the emulsion behaves as a sustained release reservoir for the drug as shown from the in vitro release data in Example 16.

Example 18

Evaluation of efficacy of paclitaxel emulsion

을 참고 조제물로서 사용하였다. 종양세포를 피하에 투여하고 지시된 복용스케쥴로 종양투여후 4일에 꼬리정맥 주입으로 치료를 시작하였다. 효능을 수명의 퍼센트 증가(% ILS)로 나타냈다. The paclitaxel emulsion of Example 6 was evaluated for efficacy on stage B16 melanoma tumors in hairless mice and the data is shown in Table 3. Once Again Commercial Product Taxol

Was used as reference formulation. Tumor cells were administered subcutaneously and treatment started with tail vein infusion 4 days after tumor administration with the indicated dosing schedule. Efficacy is expressed as a percentage increase in lifespan (% ILS).

을 투여함으로써 얻어졌고, b)%ILS 값은 30, 40 또는 50mg/kg Q2Dx4로 파클리탁셀의 α-토코페롤 에멀젼을 투여함으로써 30 내지 50%로 증가하였으며, c)에멀젼이 30, 50 및 70mg/kg Q4Dx3로 투여될 때 좋은 복용반응이 관찰되되며 더불어 70mg/kg에서 약 80% ILS가 관찰되며, d)4일에 단 한번 복용된 90mg/kg에서 조차 약 36% ILS였다. 이들 데이터는 본 발명의 에멀젼이 파클리탁셀의 효능을 실질적으로 개선시킬 가능성을 명백하게 나타낸다.The following results are derived from the data in Table 3: a) Taxol increased lifespan of about 10% to 10 mg / kg Q2Dx4.

B)% ILS values were increased to 30-50% by administration of α-tocopherol emulsion of paclitaxel with 30, 40 or 50 mg / kg Q2Dx4, c) emulsions of 30, 50 and 70 mg / kg Q4Dx3 A good dose response was observed when administered as well as about 80% ILS at 70 mg / kg and d) about 36% ILS even at 90 mg / kg taken only once every 4 days. These data clearly show the possibility that the emulsion of the present invention substantially improves the efficacy of paclitaxel.

Example 19

Evaluation of efficacy of paclitaxel emulsion

을 참고 조제물로서 다시 사용하였다. 실시예 18의 것과 본질적으로 동일한 방법을 사용하였다. 이 연구로부터의 데이터를 표 3에 요약하였다. 효능을 다음과 같이 나타내었다: a)퍼센트 종양성장 억제(%T/C, 여기에서 T 및 C는 각각 처리동물 및 대조동물을 나타낸다); b)종양성 장 지연값(T-C), 및 c) 3.32x 종양 배가 시간에 대한 T-C 값의 비로서 정의되는 Log 세포 죽임(log cell kill). 이 특이한 종양모델에 대한 후자의 패라미터는 1.75일이 되도록 계산되었다. 표 4의 결과로부터 알 수 있듯이, 모든 효능의 측정: 종양 성장억제, 종양 성장 지연값 및 Log 세포 죽임은 특히 에멀젼이 매 4일마다 70mg/kg으로 복용될 경우 Taxol

에 대한 약물송달 부형제로서 α-토코페롤 에멀젼의 우수한 효능을 나타낸다. 실시예 16에서 설명된 바와 같이, 이 증가된 효능은 개선된 약물 생체적합성 및/또는 유지된 방출의 결과일 것이다.The emulsions of Examples 6, 7 and 8 (QWA, QWB and QWC, respectively) were compared for efficacy on B16 melanoma in mice; Taxol

Was used again as a reference formulation. Essentially the same method as in Example 18 was used. Data from this study is summarized in Table 3. Efficacy was expressed as follows: a) percent tumor growth inhibition (% T / C, where T and C represent treated and control animals, respectively); b) tumor cell delay value (TC), and c) log cell kill, defined as the ratio of TC values to 3.32 × tumor doubling time. The latter parameter for this unique tumor model was calculated to be 1.75 days. As can be seen from the results in Table 4, all the measures of efficacy: tumor growth inhibition, tumor growth retardation value and log cell killing were especially taxol when the emulsion was taken at 70 mg / kg every 4 days.

It shows good efficacy of α-tocopherol emulsion as a drug delivery excipient for. As described in Example 16, this increased efficacy would be the result of improved drug biocompatibility and / or sustained release.

^a SEM = standard error of the mean

^b % ILS =% increase in lifespan = [(TC) / C] x100 where:

T = average survival of treated animals                 

C = average survival of control animals

ILS values greater than 50% according to NCI standards indicate significant anti-tumor activity.

Tumor doubling time is calculated to be 1.75 days.

% T / C = tumor growth inhibition (15 days) = (median weight of tumor in control group / median weight of control group) X 100

T-C = tumor growth retardation value = medium time between treatment (T) and control (C) tumors reaching a predetermined size (always 750-1000 mg).

Log cell kill = (T-C value) / (3.32x tumor doubling time)

Example 20

Self-emulsification of α-tocopherol / Tagat TO mixture

2.0 gm of α-tocopherol and 800 mg of Tagat TO (Goldschmidt Chemical Corp, Hopewell VA) were dissolved together. About 80 mg of the oily mixture was transferred to a test tube and water was added thereto. Upon moderately hand mixing, a rich milky emulsion occurred immediately, consistent with the “self-emulsifying system” presented as a drug delivery system in which the surfactant-oil mixture spontaneously forms an emulsion upon exposure to an aqueous medium.

Example 21

Self-emulsifying formulations containing paclitaxel

Paclitaxel 50 mg / ml was prepared in α-tocopherol by the method described in Example 1. Tagat 20% (w / w) was added. The resulting mixture was clear, viscous and amber. The amount of 100 mg of the oily mixture was transferred to a test tube. With vortex mixing, a fine emulsion was obtained while adding 1 mL of water.

Example 22

Self-emulsifying composition of paclitaxel

Paclitaxel 50 mg / ml was prepared in α-tocopherol by the method described in Example 1. After ethanol was removed under vacuum, 20% TPGS and 10% polyoxyethylene glycol 200 (Sigma Chemical Co) were added by weight. Self-emulsification of the system was then performed by adding 20 mL of deionized water to 100 mg of the oily mixture at 37 ° C. During moderate mixing, a thin emulsion was formed with a Malvern Mastersizer (Malvern Instruments, Worcester MA) consisting of fine emulsion particles with an average particle diameter of 2 microns, and 90% of the 10 cumulative distribution showing less than 10 microns.                 

Example 23

Etoposide Emulsion Formulation of α-tocopherol

4 mg of etoposide (Sigma Chemical Co) was dissolved in the following surfactant-oil mixture:

Etoposheet 4 mg

α-tocopherol 300 mg

TPGS 50 mg

Poloxamer 407 50 mg

Ethanol and mild warming were used to form a clear amber drug solution in oil. Ethanol was then removed under vacuum.

A pre-emulsion was formed by adding 4.5 mL of water containing 4% sorbitol and 100 mg TPGS using sonication at 45 ° C. Particle size was further reduced by treatment with Emulsiflex 1000 (Avestin, Ottawa Canada). The Emulsiflex 1000 body was assembled with a pair of 5 mL syringes and the entire equipment was heated to 45 ° C. prior to use. The 5 mL of emulsion was then passed through it approximately 10 times by hand. Free flowing, practical etoposide emulsions in α-tocopherol excipients were obtained.

The inventors note that etoposide in dissolved form in α-tocopherol can also be used as oral dosage form by fitting the methods of the above examples.

Example 24

Dissolution of Ibuprofen or Griseofulvin in α-tocopherol

Ibuprofen is an analgesic and can be administered by injection if necessary if there is a risk of irritating the stomach. The following ibuprofen solution in α-tocopherol can be anesthetized for intravenous administration.

12 mg of crystalline Ibuprofen (Sigma Chemicals) was dissolved by moderate heating in 120 mg of α-tocopherol without solvent. The resulting 10% ibuprofen solution in vitamin E can be emulsified by the method described in Examples 4, 6, 7, 8 or 22.

12 mg of the antifungal compound griseofulvin was first dissolved in 3 mL of anhydrous ethanol; 180 mg of α-tocopherol was added and ethanol was removed with moderate heating in vacuo. The resulting griseofulvin solution in α-tocopherol is transparent and can be emulsified by the method described in Examples 4, 6, 7, 8 or 22.

Example 25

Vitamin E Succinate Emulsion Formulation

Vitamin E succinate has been suggested as a therapeutic for the treatment of lymphoma and leukemia and for the chemoprevention of cancer. The following are compositions and methods for the emulsification of vitamin E in α-tocopherol. Sucrose ester S 1170 is a product of Mitsubishi Kagaku Foods Corp, Tokyo Japan. Vitamin E succinate as a free acid was purchased as a white powder from ICN Biomedicals, Aurora, OH. Other surfactants, such as Pluronic, and emulsions comprising TPGS together with α-tocopherol and α-tocopherol succinate can be prepared in a similar manner, with or without therapeutic agents.                 

8 gm of α-tocopherol and 0.8 gm of vitamin E succinate were dissolved together in ethanol in a round bottom flask. After the solvent was removed, 100 mL of aqueous buffer was added. The alkaline buffer consists of 2% glycerol, 10 mM trimethanolamine, and 0.5 gm% sucrose ester Sl170. After mixing for 2 minutes, the preemulsion was transferred to an Avestin Model C-5 homogenizer and homogenized for about 12 minutes at a process feed temperature of 58 ° C. The pressure difference across the interaction head was 25 to 26 kpsi. The pH was carefully observed during homogenization and adjusted to pH 7.0 as required. Care was taken to shut off oxygen during the process. A fine white emulsion was obtained.

Example 26

Α-tocopherol levels in esters

Commercially available esters: The levels of α-tocopherol in tocopherol-acetate, -succinate, nicotinate, -phosphate and TPGS were provided by the dealer or measured by HPLC. The free α-tocopherol concentrations in these solutions were below 1.0%, generally below 0.5%.

Example 27

Resveratrol Emulsion Formulation

Resveratrol is an extract of grape skins and is the first chemopreventive agent for cancer. This has been proposed as a dietary supplement.

Resveratrol was purchased from Sigma Chemical Co. Although it is poorly soluble in ethanol, 10 mg of resveratrol, 100 mg of α-tocopherol, 100 mg of TPGS and ethanol form a clear solution rapidly. Removal of ethanol left a clear amber oil.

The oily solution of Resveratrol can be formulated as a self-emulsifying system for oral delivery by various methods of the above examples.

Example 28

Muramyl Dipeptide Preparation

Muramil dipeptides are derived from mycobacteria and are potent immunostimulants that represent Muramil peptide mycolic acid and lipopolysaccharides. For example, they are used in the treatment of cancer, in particular in connection with anticancer vaccines, by stimulating the immune system to target cancer. More recently, the synthetic analogue muroctacin has been proposed to reduce nonspecific side effects of bacterial wall extracts.

N-acetylmurayl-6-O-steroyl-1-alanyl-d-isoglutamine was purchased from Sigma Chemical Co. and 10 mg was dissolved in 100 mg α-tocopherol and 80 mg TPGS. Ethanol was used as a cosolvent to aid dissolution of the dipeptide and was removed by distillation under vacuum to give a clear solution in α-tocopherol and surfactant.

This oil solution of the drug can be emulsified for parenteral administration by the various methods of the above embodiments.

Example 29

Alcohol-containing emulsions

In an attempt to adapt the teachings of PCT WO 95/11039 to oral administration of paclitaxel, the following preparations were made.

Paclitaxel 0.125 gm

α-tocopherol 0.325 gm

TPGS 0.425 gm

Ethanol 0.125 gm

As before, paclitaxel was dissolved in α-tocopherol and TPGS using ethanol and then ethanol was removed under vacuum. Residual ethanol by dry weight was less than 3 mg (0.3% w / w). Then 0.125 gm of fresh anhydrous ethanol was backfilled to the preparation. After mixing, formulation suitability for oral administration as gelatin capsules was simulated by the following test. 100 mg portion of free-flowing oil was added to 20 mL of water at 37 ° C. and mixed gently using a vortex mixer. A fine emulsion was obtained. However, after 20 minutes microscopy showed a large number of crystal growths in the rosette, which is characteristic of paclitaxel precipitation. Since a large amount of drug enters the duodenum in crystalline form and in the duodenum it is not ingested due to its physical form, it was concluded that this composition is not suitable for oral administration of paclitaxel. The inventors believe that excess ethanol, together with the high ratio of TPGS to α-tocopherol, is responsible for the crystallization of the drug observed from this preparation.

Example 30

Alcohol-containing α-tocopherol emulsion

In an attempt to adapt the teachings of PCT WO 95/11039 to intravenous administration of paclitaxel, the following preparations were made.

Paclitaxel 0.050 gm

α-tocopherol 0.100 gm

0.200 gm of lecithin

0.100 gm of ethanol

Butanol 0.500 gm

As before, paclitaxel was dissolved in α-tocopherol and TPGS using ethanol and then ethanol was removed under vacuum. Residual ethanol by dry weight was less than 2 mg (0.5% w / w). Then 0.100 gm of fresh anhydrous ethanol and 0.500 gm of n-butanol were backfilled to the preparation. A clear oil was obtained. Injectable concentrates were tested for biocompatibility in administration by standard pharmaceutical practice of mixtures with saline. About 200 mg of oil was added to 20 mL of brine and mixed. Large flakes of insoluble material were produced immediately and the largest amount of material formed thick deposits on the test tube walls. The mixture was clearly unsuitable for parenteral administration by any route, and the inventors therefore believe that it is independent of the identity of the drug contained in the preparation. After trial and error, the inventors found that lecithin is a poor choice as a surfactant for α-tocopherol because of its low HLB (about 4). Other successful examples described herein for fine emulsions suitable for parenteral administration have all been made using surfactants of high HLB. These surfactants include TPGS (near HLB 17), poloxamer 407 (HLB about 22) and Tagat TO (HLB about 14.0). Overall, the inventors have found that α-tocopherol emulsification is best performed using a main surfactant greater than HLB> 10, preferably 12. Lecithin is not in this class, although it can be used as a cosolvent. By comparison, typical o / w emulsions of triglycerides are prepared with surfactants of HLB 7 to 12, which is an α-tocopherol emulsion, also a factor in which the solubility of lipophilic and mild polar lipophilic drugs in α-tocopherol is preferred. It is shown to be the only class due to the polarity and extreme hydrophobicity of α-tocopherol. Emulsions: Theory and Practice , 2nd Ed. p. 248 (1985).

Example 31

Various formulations useful in the invention (Table 5) are prepared as follows:                 

Formulation A-Split Surfactant

1) 1.25 g TPGS and 1.01 g Pluronic F127 were dissolved in 79.00 g of water for injection by heating and stirring.

2) 0.533 g paclitaxel was dissolved in 6.354 g PEG400 by mixing (low shear) at 75 ° C.

3) 3.992 g TPGS and 8.490 g Vitamin E were added and mixed (low shear) at 45 ° C. until the TPGS melted and the mixture appeared homogeneous. This oil phase is slightly excess in view of the incomplete migration to stage 4.                 

4) The aqueous phase (step 1) was heated to 45 ° C. and the 45 ° C. oil phase (step 2 + 3) was poured over 1 minute and mixed at intermediate shear (experimental mixing motor). Mixing was continued for at least 2 minutes to form a crude emulsion.

5) The emulsion was homogenized in Avestin C5 in continuous recycle mode for 1 hour at 22 Kpsi peak stroke pressure.

6) The actual amounts and percentages shown in the table correct for incomplete movement of the oil phase during step 4.

Such methods using split surfactants are useful where the solubility of certain surfactants in the oil phase is limited.

Formulation B-all surfactants in oil phase

1) 1.066 g paclitaxel was dissolved in 12.887 g PEG400 by mixing (low shear) at 75 ° C.

2) 10.739 g TPGS and 2.157 g Pluronic F127 were added and mixed (low shear) at 50-60 ° C. until both surfactants were completely melted / dissolved.

3) 17.176 g Vitamin E was added and mixed (low shear) at 45-50 ° C. until the mixture appeared homogeneous.

4) 21.8 g of the oil phase produced in steps 1-4 were added to 79.5 g water over 1 minute while mixing at intermediate shear (experimental mixing motor). Mixing continued for a total of 3 minutes to form a crude emulsion.

5) The emulsion was homogenized in Avestin C5 in 30 minutes continuous recycle mode at 22 Kpsi peak stroke pressure.

From the overall point of view of the process, there is an advantage of having all the surfactants in the oil phase. Dispersion and subsequent homogenization of the pre-emulsion are both facilitated and potential gelling of high melting point surfactants such as TPGS can be avoided.

Example 32

Etoposide Emulsion

Vitamin E emulsion (6.0% vitamin E, 3.5% TPGS, 6.0% PEG400, 8% Pluronic F-127) containing 2 mg / ml etoposide was prepared as follows:

1) 0.1044 g of Etoposide was dissolved in 3.1435 g of PEG400 (5 min at 65 ° C.).

2) 2.0447 g TPGS and 3.1563 g vitamin E were added and mixed until completely dissolved.

3) The oil phase is mixed with 42.4 g of water for injection, which contains 0.5 g of Pluronic F-127 at 44 ° C. (the water phase is boiled and degassed before mixing with the oil phase) and the pre-emulsion is formed by simple sonication. It was.

4) Homogenized in Avestin CS at 22-24 Kpsi to form a fine emulsion.

Example 33

Etoposide Emulsion

Α-tocopherol emulsions containing PEG300 and 2 mg / ml Etoposide were prepared as follows.                 

Etoposide was first dissolved in PEG300 (10 min at 72 ° C.). Next, TPGS and vitamin E were added to the drug solution. The water phase (WFI containing Poloxamer 407) was boiled and degassed before use. The pre-emulsion was prepared by adding 5 g of oily phase to 45 g of water at 45 ° C. After mixing for 3 minutes, the pre-emulsion was homogenized at 25 Kpsi for 30 minutes to produce a fine emulsion. The final composition of the emulsion is shown below:

ingredient Composition (%, w / w)

Etoposide 0.2

Vitamin E 3.0

TPGS 1.5

PEG300 3.0

Poloxamer 407 1.0

Injection Water (WFI) 92.3

Example 34

Additional injectable paclitaxel emulsions are shown in Table 6.                 

Example 35

The compositions of the various self-emulsifying emulsions of the present invention are shown in Table 7.                 

The emulsions described in Table 7 were synthesized as follows.

SEFP-1

Paclitaxel and PEG 400 were heated together at 60-67 ° C. and stirred until the drug dissolved in PEG (15 minutes). Next, TPGS and Pluronic F127 were added and stirred at 70 ° C. for 10 to 15 minutes to dissolve the surfactant. Finally, vitamin E (α-tocopherol) was added and mixed for 5-10 minutes at 55 ° C. until the mixture became clear and homogeneous. Dilution with water phase gives a fine emulsion.

SEFP-2

Paclitaxel and PEG400 were first stirred at 65-75 ° C. for 45 minutes, TPGS was added and stirring was continued for another 30 minutes to completely dissolve all three components to produce a clear solution. Finally, Solutol HS-15 and Vitamin E were added and mixed at 55 ° C. for about 5 minutes to obtain a clear homogeneous liquid. Dilution with water phase gives a fine emulsion.

Example 36

The self-emulsifying emulsion composition of further paclitaxel is shown in Table 8.

SEFP-3 and SEFP-4 were first prepared by dissolving paclitaxel in Solutol HS-15 + PEG400 by low shear mixing at 60-70 ° C. (<30 min), followed by addition of TPGS and α-tocopherol, and briefly mixing To form a clear solution. At room temperature TPGS solidification can be observed, but at 37 ° C. remains a clear liquid.

The particle size of the emulsion was determined by diluting SEFP-3 and SEFP-4 as follows: Phosphate buffered saline at 37 ° C. by low shear mixing of 0.2 ml of SEFP-3 or SEFP-4 using a stirring bar for 5 minutes. Dilute to 100 ml. The emulsion formed quickly and its particle size was measured by Malvern Mastersizer. It was found that the volume average diameters of SEFP-3 and SEFP-4 were 2.49 and 1.55 mu m, respectively.

The average droplet diameter of the resulting emulsion for an effective self-emulsifying system should be less than 10 μm, preferably less than 5 μm.

Example 37

Paclitaxel Emulsion with PEGylated Phospholipids

DMPE-PEG ₂₀₀₀ (dimyristoyl phosphatidyl ethanolamine-polyethylene glycol 2000) with emulsion was prepared (Table 9). When present, Paclitaxel was first dissolved in PEG400 by low shear mixing at 75 ° C. Other components were added and simply mixed (after melting TPGS, and P 407 for DMPEG-2) to form a clear solution. Vacuum was applied to remove the gas from the oil phase prior to emulsification and bring the oil phase to 45 ° C. The water was boiled for 15 minutes to remove the gas and then reached 45 ° C. The two phases were mixed at 45 ° C. at low to medium shear to form a pre-emulsion. For preparation DMPE-PEG-P2, DMPE-PEG-P3 and DMPE-PEGP-4, this was achieved by simply adding warm water on the oil and stirring by hand while sonicating. The pre-emulsion for DMPE-PEG2 was prepared by pouring an oil phase into water while stirring with an experimental mixing motor. The pre-emulsion was immediately homogenized in an Avestin C5 homogenizer at 20-22 Kpsi peak stroke pressure to produce a fine emulsion with an average droplet diameter of less than 200 nm and a 99% cumulative distribution.

Example 37

Efficacy Data

Formulation D (Table 6) was evaluated for efficacy on B16 melanoma in mice as described in Examples 18 and 19, and this data is summarized in FIG. 5. Comparative efficacy data is shown in Table 10.                 

% T / C = tumor growth inhibition (treatment median tumor weight / control median tumor weight) × 100

T-C = tumor growth retardation value (medium time for treatment (T) and control (C) tumors to reach a predetermined size (> 750 mg))

Log cell kill = (T-C value) / (3.32 x tumor doubling time) Tumor doubling time is calculated to be 1.75 days

을 능가하는 조제물 D의 중대한 개선을 표시한다.Consistent with the data in Table 4, the evaluation of efficacy by tumor growth inhibition, tumor growth retardation, and Log cell killing was assessed by Taxol.

Indicates a significant improvement in Formulation D that surpasses.

Example 38

Physical stability data

The physical stability of Formulation D was assessed by the potential particle size change upon storage and this data is shown in Table 11.                 

Particle size was measured using a Nicomp 370 sub-micron particle size analyzer. As can be seen from the data in Table 11, no significant change was observed in the average droplet diameter or 99% cumulative distribution of the particles. The latter parameter is mainly used as an indicator of particle aggregation and growth. In addition, no precipitation or other large changes were observed during storage. Long term stability is in progress.

Example 39

Chemical stability

The chemical stability of Formulation D (Table 6) was monitored by HPLC using the procedure of Example 5 and this data is shown in Table 12. HPLC is used to quantify the concentration of paclitaxel and digests. In Table 12, drug concentrations are equivalent to drug efficacy.                 

It is evident from this data that the drug efficacy of Formulation D remains unchanged under these storage conditions.

In addition, no degradation of the drug was observed during this storage time.

Example 40

Emulsions Containing PEG300 or NMP

Α-tocopherol emulsions containing PEG300 or NMP (N-methyl-2-pyrrolidone) and including 10 mg / ml paclitaxel are shown in Table 13.                 

In both cases, paclitaxel was first dissolved in solvent (PEG300 or NMP) with low shear mixing. The use of PEG300 was heated to 60 ° C. to speed up dissolution, but the preparation using NMP dissolved the drug sufficiently in a few minutes at room temperature. The remaining ingredients (except water) were then added and the mixture was heated to 60-65 ° C. with low shear mixing to melt the solid surfactant and produce a homogeneous clear solution. The solution was brought to 45 ° C. and then 45 ° C. water was added to it. The resulting mixture was treated under medium shear to produce a thick white crude emulsion that was very similar in appearance to the pre-emulsion of Formulation D (Table 6). These emulsions can be further homogenized at high pressure to produce fine emulsions.

Example 41

Large scale preparation of preparation D (QW8184)

Using a procedure similar to those described in the previous examples, Formulation D (Table 6) was prepared on a large scale in 2 × 2L sub-lots having the following composition (shaded areas indicate the oil and water content of the emulsion) .

For the preparation of the pre-emulsion, 416.8 g of oil phase of sub-lot 1 and 413.2 g of oil phase of sub lot 2 were mixed with 1580 g of water for injection (5 min at 46 ° C.). Homogenization produced a fine emulsion having an average droplet diameter of about 70 nm, very similar to the average droplet diameter of Formulation D at small scale (Table 11). This scaled preparation was filtered through a 0.2 micron filter to further sterilize.

Example 42

Evaluation of Hemolytic Activity of Drug-Free Emulsions

Large scale (2.5 L) of Formulation D in the absence of paclitaxel was prepared as described in Example 41 with the following composition.                 

For the preparation of the pre-emulsion, 496.7 g of oil phase was mixed with 2000 g of water for injection (5 min at 46 ° C.). Homogenization and Filter Sterilization and this preparation was assessed for total hemolytic response with human blood using the following procedure.

Healthy blood of volunteers was collected using heparin with the Vacutainer rod. Plasma was initially pale yellow and negative for hemolysis. Several drops of whole blood and drug-free emulsion were put together under the cover glass and observed under a microscope for several minutes. While in contact, red blood cells (RBCs) remained normal red blood cells. No apparent agglomeration of the emulsion particles was seen. No gross change in platelet or WBC form was noted. Next, whole blood and excipients were mixed at 1: 1 and 5: 1 (v / v) in test tubes. As a control, whole blood was mixed 1: 1 with saline for injection. All mixtures were incubated at 37 ° C. and tested at 10 and 30 minutes. The supernatant of all three tubes was pale yellow and clear. It can be concluded from this study that there is no immediate overall hemolytic response between the emulsion excipient and the blood. Unlike some reports in the literature on surfactant-induced hemolysis of RBCs, this suggests that the morphology of the red blood cell membrane is not disturbed by the surfactants present in the emulsion.

Example 43

Physical stability data

Table 16 shows the long term stability of the large scale preparation of Example 41 upon storage at 4 ° C. or 25 ° C. for 9 months. This demonstrates that at least during this storage time the average droplet diameter and 99% cumulative distribution did not change significantly from its initial values of about 65 and 150 nm, respectively, and this emulsion is still within the scope of the specification.

Example 44

Chemical stability

The nine month chemical stability data of the large scale preparation of Example 41 regarding paclitaxel efficacy and known degradation levels are shown in Tables 17 and 18. As can be seen from these results, there was no significant change in the level of drug efficacy or known degradation products, and the product is still within the scope of the specification at both storage temperatures.

Example 45

Efficacy Assessment

The formulation of Example 41 was evaluated for efficacy against B16 melanoma as described in Examples 18, 19 and 37, and the results are summarized in Table 18.                 

a:% T / C = (median tumor weight of treatment group / median tumor weight of control group) x 100

b:% TGI = 100-(% T / C)

c: T-C = tumor growth retardation value (medium time between treatment (T) and control (C) reaching a defined size (> 750 mg))

d: Log cell kill = (T-C value) / (3.32 x tumor doubling time)

의 MTD를 포함하거나 또는 상당히 초과하지만 내성이 있는 투여량으로 마우스에서 우수한 항종양 활성을 나타냈다. 그러한 효과는 이전의 주사가능한 파클리탁셀 에멀젼에 대해서는 보고되지 않았다. MTD는 정확한 독성 연구로부터 측정된 최대 내성 투여량이다.To the end of all efficacy, QW8184 is a Taxol

Doses containing or significantly exceeding MTD of showed good antitumor activity in mice. Such effects have not been reported for prior injectable paclitaxel emulsions. MTD is the maximum tolerated dose measured from exact toxicity studies.

Example 46

Efficacy Assessment

을 사용하였다. 무모마우스 숙주에서 피하 적으로 성장한 종양으로부터 수집된 IGROV-1 인간 난소 암종의 단편을 무모마우스에 투관침에 의해 피하적으로 이식하였다. 종양이 대략 5 x 5mm 크기였을 때, 동물을 귀에 꼬리표가 달린 종양을 지니는 마우스가 군 당 9 마리씩 함유된 처리 및 대조군으로 나누었다. QW8184를 20, 40 및 60mg/kg으로 q3dx5, q4dx5 및 qdx5 스케쥴에 따라 정맥내 투여하였다. Taxol

을 최대 내성 투여량인 20mg/kg으로 동일한 스케쥴에 따라 정맥내 투여하였다. 매주 2번 마우스의 중량을 재고, 종양 측정은 1일을 시작으로 캘리퍼스에 의해 행하고, mg 종양 중량으로 환산했다. 대조군 종양이 대략 1gr에 도달했을 때 실험을 종료하고, 종양을 절제하여 중량을 재고 군 당 평균 종양 중량을 계산하였다. 이 데이타를 표 20에 요약한다.The antitumor activity of QW8184 (Example 41) against human ovarian tumor xenograft IGROV-1 was evaluated using the prototype Taxol as a reference preparation.

Was used. Fragments of IGROV-1 human ovarian carcinoma collected from subcutaneously grown tumors in a hairless mouse host were implanted subcutaneously into trocars by trocars. When tumors were approximately 5 × 5 mm in size, animals were divided into treatments and controls containing 9 mice per group with tumors with ear tags. QW8184 was administered intravenously at 20, 40 and 60 mg / kg according to the q3dx5, q4dx5 and qdx5 schedules. Taxol

Was administered intravenously according to the same schedule at the maximum tolerated dose of 20 mg / kg. Mice were weighed twice a week, and tumor measurements were performed by calipers starting at 1 day and converted to mg tumor weight. The experiment was terminated when the control tumors reached approximately 1 gr, the tumors were excised and weighed to calculate the average tumor weight per group. This data is summarized in Table 20.

의 투여는 2개 스케쥴 에 대해 3 마리의 완전 종양 반응을 가져왔다. qdx5 스케쥴에 대해서, QW8184 및 Taxol

의 항종양 활성은 유사하였다. 그러나, QW8184는 중독성 사망이 없음에 따라 더 내성이 있었고, 반면 Taxol

은 6 마리의 중독성 사망이 기록되었다. QW8184는 투여 스케쥴과 관계없이 투여량-의존 방식에서 IGROV-1 인간 난소 이종이식편 모델에 대해 매우 활성이었고, Taxol

보다 더 내성이 있었다. Administration of QW8184 at 20, 40 and 60 mg / kg according to the q3dx5 or q4dx5 schedule resulted in nearly 100% tumor growth inhibition at all doses, with 2, 8 and 7, and 3, 9 and 8 complete tumors, respectively Have a response. By comparison, Taxol

Administration of 3 resulted in 3 complete tumor responses to the two schedules. For the qdx5 schedule, QW8184 and Taxol

The antitumor activity of was similar. However, QW8184 was more resistant as there was no addictive death, whereas Taxol

6 addictive deaths were recorded. QW8184 was very active against the IGROV-1 human ovarian xenograft model in a dose-dependent manner regardless of the dosing schedule, and Taxol

More resistant than

Example 47

Pharmacokinetic Studies

을 사용하여 측정하였다. 약물을 3시간 주입(Taxol

) 또는 거환 투여(QW8184)로서 수컷 또는 암컷 래트에 정맥내 투여하였다. 혈액 샘플을 투여량 투여 후 0 내지 72시간 수집하고, 혈장을 원심분리에 의해 제조하고 LC/MS/MS 검출기를 갖는 고성능 액체 크로마토그래피(HPLC)법을 사용하여 파클리탁셀 농도에 대해 분석하였다. 약물동태학 분석을 모델 독립적 방법을 사용하여 평균 복합 혈장 농도-시간 프로필에 대해 수행하였다. 유도된 약물동태학 파라미터를 표 21에 나타낸다. 측정된 약물동태학 파라미터는 다음과 같았다:The pharmacokinetics of the formulation of Example 41 (QW8184) as Taxol as a reference formulation upon single intravenous administration of 10 mg / kg

Measured using. 3 hours of drug injection (Taxol

) Or male or female rats intravenously (QW8184). Blood samples were collected from 0 to 72 hours after dose administration and plasma was prepared by centrifugation and analyzed for paclitaxel concentration using high performance liquid chromatography (HPLC) with LC / MS / MS detector. Pharmacokinetic analyzes were performed on average complex plasma concentration-time profiles using model independent methods. The derived pharmacokinetic parameters are shown in Table 21. The pharmacokinetic parameters measured were as follows:

T _max : time required to reach maximum plasma level (C _max )

C _max : maximum plasma concentration of the drug

AUC _0-t : non-extrapolated region under plasma concentration-time curve from time 0 to end time t of plasma sample collection

AUC _0-∞ : Extrapolation region under plasma concentration-time curve from time 0 to infinity

K _e : Removal rate constant

T _1/2 : elimination half-life

V _d : distribution volume

CL: plasma clearance

V _ss : Distribution volume at steady state

의 정맥내 주입 후의 상응하는 값보다 상당히 높았다. 혈장에서 파클리탁셀의 말기 T_1/2는 2개 처리에 대해 유사하였다. 조직 결합은 정상상태에서의 분포 부피(V_ss)의 차이로부터 표시되는 바와 같이 QW8184 보다 Taxol

에서 더 광범위하였다. 파클리탁셀의 약물동태학 파라미터에서의 중대한 차이는 수컷 및 암컷 동물간에 관찰되지 않았다. Both C _max and AUC _0-∞ values after QW8184 intravenous bolus administration

Was significantly higher than the corresponding value after intravenous infusion. Terminal T _1/2 of paclitaxel in plasma was similar for the two treatments. Tissue binding is more taxol than QW8184 as indicated from the difference in distribution volume (V _ss ) at steady state.

More extensive in No significant difference in pharmacokinetic parameters of paclitaxel was observed between male and female animals.

Example 48

Tocotrienol Emulsion of Paclitaxel for Intravenous Administration

A tocotrienol emulsion containing the following 5 mg / mL paclitaxel was prepared, which is suitable for intravenous cancer therapy in mice.

토코트리엔올 농축물을 Golden Jomalina Food Industries (Kuala Lumpur, Malaysia)로부터 입수했다. 붉은 빛을 띤 갈색 오일의 HPLC 분석은 4개의 주요한 피크를 나타냈다. 대부분의 부분에서 약 20%의 잔류 d-α-토코페롤 함량으로 인해, 발명자들의 오일에 대한 추정 αTE는 ~0.3이다. 그것을 2부의 δ-토코페롤(Sigma Chemicals)로 희석하여 αTE를 0.1로 조정하였다. 약물 오일 용액을 우선 가열 및 초음파처리하면서 PEG-400에 파클리탁셀을 용해시킴으로써 제조하였다. 다음에, TPGS 및 토코트리엔올 오일을 가하였다. 마지막으로, poloxamer를 가하고 72℃에서 용융시켜 균질 투명한 호박색 기름을 얻었다. 혼합물을 rotevap에서 진공하에 기체를 제거하고, 사용할 때까지 45℃에 유지하였다. Gold tri-e

Tocotrienol concentrates were obtained from Golden Jomalina Food Industries (Kuala Lumpur, Malaysia). HPLC analysis of a reddish brown oil showed four major peaks. Due to a residual d-α-tocopherol content of about 20% in most parts, the estimated αTE for the inventors' oil is ˜0.3. It was diluted with 2 parts δ-tocopherol (Sigma Chemicals) to adjust αTE to 0.1. Drug oil solutions were prepared by dissolving paclitaxel in PEG-400 first with heating and sonication. Next, TPGS and tocotrienol oil were added. Finally, poloxamer was added and melted at 72 ° C. to obtain a homogeneous transparent amber oil. The mixture was degassed under vacuum at rotevap and kept at 45 ° C. until use.

An aqueous phase consisting of 40 mL of 5 mM citrate TEA buffer (pH 6.8) was brought to 45 ° C. prior to addition. Upon addition the resulting mixture was mixed vigorously to liberate any sticky oil on the flask wall. This suspension was then placed in a feed vessel and treated in a C5 homogenizer for 10 minutes with continuous recycling. Treatment conditions were 45 ° C. feed temperature, 20 Kpsi treatment pressure, 120 mL / min flow rate. A heat exchanger set at 22 ° C. was placed in the outlet port to remove excess heat generated in the homogenizer. The temperature of the feed vessel was measured at 44 ° C. during steady state homogenization.

After treatment, the product was collected and cooled to room temperature. The emulsion was then aseptically filtered through a 0.2 um filter at the end, and the emulsion had an average particle size of ˜70 nm when measured on a Nicomp 370 photon correlation spectrophotometer. For convenience of handling Gen tamycin 15 ug / mL was added as a preservative.

As a result of its low viscosity, the use of tocotrienol enriched moiety (Gold Tri-E) is substantially more than similar emulsions made using d, l-α-tocopherol (Roche Vitamins) as an oil phase. Made easy to handle

Example 49

Δ-tocopherol emulsion of paclitaxel for intravenous administration

Small mixtures are valuable for formulation development. In this example, d-δ-tocopherol with an αTE content of ˜0.0 was purchased from Sigma Chemical in 90% purity. Drug solutions were prepared in each mixture according to the following table.

Dehydrated ethanol was first used to dissolve the drug crystals. The sample was then placed under vacuum on the rotevap until all ethanol was removed by gravimetry. All samples were cooled and examined. Each sample consisted of a paclitaxel solution in δ-tocopherol in heavy amber oil and was visually clear. The calculated paclitaxel concentration in oil is shown below.

To this oil / drug mixture 50 mg TPGS and 10 mg Poloxamer 407 were added as surfactant. 60 mg PEG-400 was added as osmolite. The mixture was dissolved while heating to form a golden oil. The αTE content of these oils is 0.08 αTE units when TPGS surfactants are optionally considered, and 0.0 αTE units when used in measurements on tocopherol oil phase alone.

This oil-drug concentrate was then cooled to 45 ° C. and 850 μl warm water was added. Micro-tip sonication horns were used to prepare pre-emulsions of crude particles. Microscopic examination showed a thick suspension in which the particles were substantially less than 10 μm in diameter, which was further processed in a homogenizer, adjusted for pH and osmotic strength, and final sterile filtration suitable for parenteral injection.

Example 50

Emulsion Formulation of Methylcarbonate Derivative of Paclitaxel (BMS-188797) for Intravenous Administration

BMS-188797 was first dissolved in ethanol, and then α-tocopherol, Myvacet 9-45 (if present), TPGS, poloxamer 407 and PEG400 were added and mixed at high temperature (about 60 ° C.). The ethanol was then removed under vacuum at high temperature to give a clear oily phase containing the drug. This was then mixed with water for injection at 45 ° C. to prepare a pre-emulsion. The final emulsion was prepared by homogenizing in an Avestin C5 homogenizer at 10-20 minutes at 19-20 Kpsi while maintaining the temperature between 35-45 ° C. The compositions of some representative emulsions are shown in the table below. The average droplet diameter and 99% cumulative distribution of these emulsions were measured to be less than 0.1 and 0.2 μm, respectively. These emulsions can be filtered and sterilized, stable at room temperature, sufficiently resistant and efficacious in animals.

Example 52

Emulsion Formulations of Clarithromycin

The macrolide antibiotic clarithromycin was purchased as a free base from Wockhardt (Delhi, India). Vitamin E succinate (VESA) was purchased from Eastman (Freeport TN). Capmul MCM from Abitec (Janesville WI); Poloxamer F127 from Parsippany NJ; PEG400 from Spectrum Chemicals (Gardenia CA); And d-δ-tocopherol was purchased from Sigma Chemic als (St Louis MO). An oil phase consisting of d-δ-tocopherol and Capmul MCM was prepared using 2 parts of δ-tocopherol. ΑTE in the oil phase is 0.0. Next, surfactant and clarithromycin were added as shown in the table below. Dry ethanol was used to dissolve the components at 70 ° C. and then ethanol was removed under vacuum.

An aqueous phase consisting of 40 mL of 5 mM citrate TEA buffer (pH 6.8) was brought to 45 ° C. prior to addition. Upon addition the resulting mixture was mixed vigorously to liberate any sticky oil on the flask wall. This suspension was then placed in a feed vessel and treated in a C5 homogenizer (Avestin, Ottawa CA) for 3 minutes with continuous recycling. Treatment conditions were 45 ° C. feed temperature, 20 Kpsi treatment pressure, 120 mL / min flow rate. A heat exchanger set at 22 ° C. was placed in the outlet port to remove excess heat generated in the homogenizer. The temperature of the feed vessel was measured at 44 ° C. during steady state homogenization.

After treatment, the product was collected and cooled to room temperature. The emulsion was then finally sterile by filtration through a 0.2 μm filter, and the emulsion had an average particle size of less than 52 nm as measured on a Nicomp 370 photon correlation spectrophotometer.

Claims (34)

(a) combining paclitaxel and polyethylene glycol to provide a first paclitaxel-containing solution; And (b) adding tocopherol / polyethylene glycol derivative, polyoxypropylene-polyoxyethylene nonionic block copolymer, and tocopherol to the first paclitaxel-containing solution to provide a second paclitaxel-containing solution A method for preparing a paclitaxel-containing solution comprising a and not removing ethanol. (a) combining paclitaxel and polyethylene glycol to provide a first paclitaxel-containing solution; (b) adding tocopherol / polyethylene glycol derivative, polyoxypropylene-polyoxyethylene nonionic block copolymer, and tocopherol to the first paclitaxel-containing solution to provide a second paclitaxel-containing solution; And (c) mixing the second paclitaxel-containing solution with the aqueous phase to form a paclitaxel-containing emulsion And a method for producing a paclitaxel-containing emulsion, the method comprising the step of removing ethanol. 3. The method of claim 2, further comprising homogenizing the emulsion. (a) combining a water insoluble therapeutic agent with a cosolvent selected from dimethyl sulfoxide, benzyl benzoate, propylene glycol, sorbitol, mannitol, polyethylene glycol, N-methyl-2-pyrrolidone and polyvinylpyrrolidone Providing a therapeutic agent-containing solution; And (b) Tocol and alpha- selected from alpha-tocopherol, delta-tocopherol, gamma-tocopherol, beta-tocotrienol, delta-tocotrienol, gamma-tocotrienol, desmethyltocotrienol, and didesmethyl tocotrienol in the first therapeutic agent-containing solution. Adding a surfactant comprising tocopherol polyethylene glycol succinate to provide a therapeutic / tocopherol-containing solution And a method of preparing a therapeutic agent / tocopherol-containing solution, the method comprising the step of removing ethanol. The method of claim 4, wherein (c) mixing the therapeutic / tocopherol-containing solution with the aqueous phase to form a pre-emulsion; And (d) homogenizing the pre-emulsion to form an emulsion Method further comprising a. The method of claim 4 or 5, wherein the therapeutic agent is paclitaxel. delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete

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