HK1161069A

HK1161069A - Sustained-release drug carrier composition

Info

Publication number: HK1161069A
Application number: HK12101469.6A
Authority: HK
Inventors: 诺姆‧伊曼纽尔; 摩西‧纽曼; 沙洛莫‧巴拉克
Original assignee: 波利皮得有限公司
Priority date: 2008-07-14
Filing date: 2009-07-14
Publication date: 2012-08-24

Abstract

The present invention provides compositions for extended release of an active ingredient, comprising a lipid-saturated matrix formed from a biodegradable polymer. The present invention also provides methods of producing the matrix compositions and methods for using the matrix compositions to provide controlled release of an active ingredient in the body of a subject in need thereof.

Description

Sustained release pharmaceutical carrier composition

Technical Field

The present invention provides a composition for extended release of an active ingredient comprising a lipid-based matrix with a biodegradable polymer. The invention also provides methods of producing the matrix compositions and methods of using the matrix compositions to provide controlled release of an active ingredient in a subject in need thereof.

Background

Lipid-based drug delivery systems are well known in the field of pharmaceutical science. Generally, they are used to formulate drugs with poor bioavailability or highly toxic drugs or both. Among the popular dosage forms that have gained acceptance are many different types of liposomes including small unilamellar vehicles, multilamellar vehicles, and many other types of liposomes; different types of emulsions including water-in-oil emulsions, oil-in-water emulsions, water-in-oil-in-water double emulsions, submicron emulsions, micron emulsions; micelles, and many other hydrophobic drug carriers. These types of lipid-based delivery systems can be highly specialized to allow targeted drug delivery or reduced toxicity or increased metabolic stability and the like. Extended release over the range of days, weeks and longer is not a feature typically associated with lipid-based drug delivery systems in vivo.

An ideal sustained release drug delivery system should exhibit kinetic and other characteristics that are readily controlled by the type and proportion of the specific excipients used. Advantageously, the sustained release drug delivery system should provide a solution of hydrophilic, amphiphilic and hydrophobic drugs.

Periodontitis

Treatment with systemic doxycycline and NSAIDs in combination has been shown to inhibit tissue damage in the gingiva of patients with chronic periodontitis. Tissue damage results from the combination of the activity of pathogenic bacteria and host Matrix Metalloproteinase (MMP) activity. The combination of antibiotic therapy and anti-inflammatory drugs inhibits both pathways. Enhanced efficacy and reduced side effects are obtained by methods that locally release these drugs in a controlled manner.

Bone augmentation

Bone diseases requiring bone augmentation (bone augmentation) include benign and malignant bone tumors, cancers located in bone, infectious bone diseases and other bone diseases involving endocrinology, autoimmunity, malnutrition, genetic factors and etiology of imbalances in bone growth and resorption. Examples are diseases such as osteosarcoma/malignant fibrous histiocytoma of bone (PDQ), osteosarcoma, chondrosarcoma, ewing's sarcoma, malignant fibrous histiocytoma, fibrosarcoma and malignant fibrous histiocytoma, giant cell tumor of bone, chordoma, lymphoma, multiple myeloma, osteoarthritis, paget's disease of the bone, arthritis, degenerative changes, osteoporosis, osteogenesis imperfecta, bone spurs, renal osteodystrophy, hyperparathyroidism, osteomyelitis, endogenesis chondroma, osteochondroma, osteopetrosis, bone and joint problems associated with diabetes.

Immediate and delayed infection is a major complication in the field of orthopedics. Reducing complications after orthopedic treatment will contribute to efficiency and success of orthopedic treatment and in some cases it will reduce mortality. There is also a need to allow treatment at the site of infection and to induce efficacy of treatment at the site of infection.

Another important aspect in the field of orthopedics (orthotics) or orthopedics (orthotics surgery) is the need to accelerate soft and hard tissue recovery during repair and regeneration.

Liposomes and biodegradable polymers in drug delivery

To date, the initial use of lipid-bound biopolymers has been considered, but this has not been successfully introduced into clinical practice.

US 3,773,919 to Boswell et al describes the use of polymers derived from alpha-hydroxycarboxylic acids including lactic acid, glycolic acid and copolymers thereof, and their use in sustained release formulations. These polymers exhibit slow biodegradability but generally have limited drug retention.

Liposomes are described in U.S. Pat. No. 4,522,803 to Lenk et al. Liposomes generally exhibit sufficient drug-delivery drug retention capacity but relatively limited in vivo half-life. Many different types of liposomes have been developed for particular applications. Examples can be found in, among others, U.S. patent nos. 5,043,166; 5,316,771 No; 5,919,480 No; 6,156,337 No; U.S. Pat. No. 6,162,462; 6,787,132 No; 7,160,554, etc.

U.S. Pat. Nos. 6,333,021 and 6,403,057 to Schneider et al disclose microcapsules having a biodegradable membrane encapsulating a gas core. The film comprises up to 75% by weight of water-insoluble lipids with a biodegradable polymer encapsulating a core filled with air or gas. The microcapsules may be non-agglomerating (non-coalescent), dry and instantly dispersible, and serve as delivery vehicles for therapeutically active agents and/or as contrast agents for imaging body organs. Microcapsules are produced by a process wherein a water-in-oil emulsion is prepared from an organic solution comprising dissolved lipids and an aqueous solution containing a surfactant. The lyophilized mixture is redispersed in an aqueous carrier and the microcapsules are dried. The presence of water throughout the procedure precludes the formation of a water-resistant, lipid-saturated matrix; as a result, these materials are subject to significant degradation in vivo.

U.S. Pat. nos. 6,277,413 and 6,793,938 to Sankaram disclose biodegradable lipid/polymer containing compositions formed by the following process: a) forming a water-in-oil emulsion from a first aqueous phase and a volatile organic solvent phase, the volatile organic solvent phase comprising a volatile organic solvent, a biodegradable polymer or copolymer that is soluble in the organic solvent, and a lipid; b) dispersing the "water-in-oil" emulsion into a second aqueous phase free of surfactant to form solvent globules, and c) removing the volatile organic solvent from the solvent globules to form the microsphere composition suspended in the second aqueous phase. The method discloses the use of an aqueous solution, excluding the formation of a water-resistant, lipid-saturated matrix.

Jang, U.S. patent 4,882,167, discloses a controlled release matrix for tablets or implants of bioactive agents produced by dry direct compression of a hydrophobic carbohydrate polymer such as ethylcellulose and a poorly digestible soluble component, i.e., a wax, such as palm wax, fatty acid material or neutral lipids. The carbohydrate polymers used are not suitable for release in the range of weeks or months after administration by injection or implantation. In addition, the composition is prepared without any solvent (aqueous or organic), precluding the formation of a homogeneous lipid-saturated matrix structure.

U.S. patent application 2006/0189911 to Fukuhira et al discloses an anti-adhesive film of a honeycomb membrane made from polylactic acid and phospholipids as biodegradable polymers. No disclosure provides improvements to films for use as delivery systems for, for example, antibiotics or NSAID drugs. In addition, the disclosed films are cast under high humidity conditions, thus precluding the formation of a water-resistant, lipid-saturated matrix; these implants therefore undergo extensive degradation in vivo.

U.S. patent application 2006/0073203 to Ljusberg-Wahren et al discloses an orally administrable composition comprising a dry mixture of a polymer, a lipid, and a bioactive agent, desirably forming particles comprising the lipid, bioactive agent, and optionally water, upon contact with water or gastrointestinal fluids. The polymers used decompose in the digestive tract during digestion, for example during a period of less than one day. Such compositions are completely unsuitable for release over a range of weeks or months after administration by injection or implantation.

The prior art does not provide such compositions: suitable for achieving sustained or programmed or controlled release from a lipid-saturated polymer matrix for periodontal or orthopedic use. The above documents do not show the use of the disclosed compositions in the delivery of NSAID compounds, antibiotic compounds or compounds for bone augmentation.

Summary of The Invention

The present invention provides a composition for the extended release of an active ingredient comprising a lipid-based matrix comprising a biodegradable polymer. The invention also provides methods of producing the matrix compositions and methods of using the matrix compositions to provide controlled release of an active ingredient in a subject in need thereof.

In one aspect, the present invention provides a matrix composition comprising: (a) a biodegradable pharmaceutically acceptable polymer associated with a first lipid having polar groups; (b) a second lipid selected from phospholipids having a hydrocarbon chain of at least 14 carbons; and (c) a pharmaceutically active agent, wherein the matrix composition is adapted to provide sustained release of the pharmaceutically agent. In particular embodiments, the polymer and phospholipid form a substantially anhydrous matrix composition.

According to a particular embodiment, the biodegradable polymer comprises a polyester selected from the group consisting of: PLA (polylactic acid), PGA (polyglycolic acid), PLGA (poly (lactic-co-glycolic acid)), and combinations thereof.

According to a particular embodiment, the first lipid having a polar group is selected from sterols, tocopherols and phosphatidylethanolamines. According to a specific embodiment, the first lipid is mixed with the biodegradable polymer to form a non-covalent association.

According to some embodiments, the second lipid comprises phosphatidylcholine. According to some embodiments, the second lipid comprises a mixture of phosphatidylcholines. According to some embodiments, the second lipid comprises a mixture of phosphatidylcholine and phosphatidylethanolamine or any other type of phospholipid.

Any type of drug molecule may be incorporated into the matrix composition for sustained and/or controlled release. According to a specific embodiment, the pharmaceutically active agent is selected from the group consisting of: antibiotics, antifungal agents, NSAIDs, steroids, anticancer agents, osteogenic factors (osteopenic factors) and bone resorption inhibitors. According to an alternative embodiment, the pharmaceutically active agent is selected from a hydrophobic agent, an amphiphilic agent or a water-soluble agent. Each possibility represents a separate embodiment of the invention.

In another embodiment, the phospholipid is a phosphatidylcholine having fatty acid moieties of at least 14 carbons. In another embodiment, the composition further comprises phosphatidylethanolamine having a fatty acid moiety of at least 14 carbons. In another embodiment, the composition further comprises cholesterol. In another embodiment, in the matrix composition of the invention, cholesterol is present in an amount of 5 to 50 mole percent of the total lipid content of the matrix composition. In another embodiment, the matrix composition is homogeneous. In another embodiment, the matrix composition is in the form of a lipid-based matrix, the shape and boundaries of which are defined by the biodegradable polymer. In another embodiment, the matrix composition is in the form of an implant.

In some embodiments, the pharmaceutically active agent is an antibiotic incorporated into the matrix composition. In some embodiments, the antibiotic has low water solubility. In another embodiment, the antibiotic is a hydrophobic antibiotic. In another embodiment, the antibiotic is an amphiphilic antibiotic. In another embodiment, the composition further comprises a non-steroidal anti-inflammatory drug (NSAID). In another embodiment, the NSAID is also incorporated into the matrix composition. In another embodiment, the NSAID has low water solubility. Each possibility represents a separate embodiment of the invention.

In a specific embodiment, the present invention provides a matrix composition comprising: (a) a biodegradable polyester; (b) a sterol; (c) phosphatidylethanolamine having a fatty acid moiety of at least 14 carbons; (d) a phosphatidylcholine having fatty acid moieties of at least 14 carbons; and (e) an antibiotic or antifungal agent. In another embodiment, the matrix composition comprises at least 50% lipid by weight. In another embodiment, the matrix composition is homogeneous. In another embodiment, the matrix composition is in the form of a lipid-based matrix, the shape and boundaries of which are defined by the biodegradable polymer. In another embodiment, the matrix composition is in the form of an implant.

According to an alternative embodiment, the antibiotic or antifungal agent is selected from a hydrophobic agent, an amphiphilic agent or a water-soluble agent. Each possibility represents a separate embodiment of the invention.

In another embodiment, the present invention provides a matrix composition comprising: (a) a biodegradable polyester; (b) a sterol; (c) phosphatidylethanolamine having a fatty acid moiety of at least 14 carbons; (d) a phosphatidylcholine having fatty acid moieties of at least 14 carbons; and (e) a non-steroidal anti-inflammatory drug (NSAID). In another embodiment, the matrix composition comprises at least 50% lipid. In another embodiment, the NSAID has low water solubility. In another embodiment, the NSAID is a hydrophobic NSAID. In another embodiment, the NSAID is an amphiphilic NSAID. In another embodiment, the matrix composition is in the form of a lipid-based matrix, the shape and boundaries of which are defined by the biodegradable polymer. In another embodiment, the matrix composition is in the form of an implant. In another embodiment, the matrix composition is homogeneous. Each possibility represents a separate embodiment of the invention.

In another embodiment, the present invention provides a matrix composition comprising: (a) a biodegradable polyester; (b) a sterol; (c) phosphatidylethanolamine having a fatty acid moiety of at least 14 carbons; (d) a phosphatidylcholine having fatty acid moieties of at least 14 carbons; and (e) an osteogenic factor or a bone resorption inhibitor. In another embodiment, the matrix composition comprises at least 50% lipid. In another embodiment, the bone resorption inhibitor has low water solubility. In another embodiment, the bone resorption inhibitor is a hydrophobic bone resorption inhibitor. In another embodiment, the bone resorption inhibitor is an amphiphilic bone resorption inhibitor. In another embodiment, the composition further comprises an NSAID. In another embodiment, the NSAID is also incorporated into the matrix composition. In another embodiment, the matrix composition is in the form of a lipid-based matrix, the shape and boundaries of which are defined by the biodegradable polymer. In another embodiment, the matrix composition is in the form of an implant. In another embodiment, the matrix composition is homogeneous. Each possibility represents a separate embodiment of the invention.

In another embodiment, the present invention provides a matrix composition comprising: (a) a biodegradable polyester; (b) a sterol; (c) phosphatidylethanolamine having a saturated fatty acid moiety of at least 14 carbons; (d) a phosphatidylcholine having saturated fatty acid moieties of at least 14 carbons; (e) an active agent; and (f) a targeting moiety capable of interacting with a surface molecule of the target cell. In another embodiment, the active agent is selected from the group consisting of: NSAIDs, antibiotics, antifungal agents, steroids, anticancer agents, osteogenic factors, and bone resorption inhibitors. In another embodiment, the polymer and phospholipid form a substantially anhydrous matrix composition. In another embodiment, the matrix composition is capable of being degraded in vivo into vesicles (vesicles) in which some or all of the mass of the released active agent is incorporated (integrated). In another embodiment, the matrix composition is capable of being degraded in vivo to form vesicles into which the active agent and targeting moiety are incorporated. Each possibility represents a separate embodiment of the invention.

In another embodiment, the matrix composition of the invention is for sustained release of a pharmaceutically active agent, wherein at least 50% of the pharmaceutically active agent is released from the matrix composition at zero order kinetics. In another embodiment, the matrix composition of the invention is for sustained release of a pharmaceutically active agent, wherein at least 60% of the pharmaceutically active agent is released from the matrix composition at zero order kinetics. In another embodiment, the matrix composition of the invention is used for sustained release of the pharmaceutically active agent, wherein at least 65% of the pharmaceutically active agent is released from the matrix composition at zero order kinetics.

In another embodiment, the invention provides a pharmaceutical composition comprising the matrix composition of the invention and a pharmaceutically acceptable excipient. In another embodiment, the matrix composition is in the form of microspheres. In another embodiment, the invention provides a pharmaceutical composition comprising the microspheres of the invention and a pharmaceutically acceptable excipient. In another embodiment, the pharmaceutical composition is in a parenterally injectable form. In another embodiment, the pharmaceutical composition is in an undissolved form. In another embodiment, the excipient is suitable for injection. In another embodiment, the excipient is suitable for infusion. Each possibility represents a separate embodiment of the invention.

In another embodiment, the matrix composition of the invention is in the form of an implant after evaporation of the organic solvent. In another embodiment, the implant is homogeneous. Each possibility represents a separate embodiment of the invention.

In some embodiments, the biodegradable polyester of the present invention is associated with the sterol via a non-covalent bond. In some embodiments, the biodegradable polyesters of the present invention are associated with sterols via hydrogen bonding.

In another embodiment, the process of producing an implant from the composition of the invention comprises the steps of: (a) producing a matrix composition in the form of a bulk material according to the method of the invention; and (b) transferring the bulk material into a mold or solid container having a desired shape.

Also provided herein are methods of making the compositions of the invention and methods of using the same.

According to another aspect, a matrix composition for sustained release of a pharmaceutical agent is produced by a process comprising: providing a first solution or dispersion of a volatile organic solvent comprising a biodegradable polymer and a first lipid having a polar group; providing a second solution or dispersion comprising a second volatile organic solvent and a second lipid, the second lipid comprising at least one phospholipid, and a pharmaceutically active agent; mixing the first and second solutions to form a homogeneous mixture; the volatile solvent is evaporated to produce a homogeneous polymeric phospholipid matrix comprising the pharmaceutically active agent. The choice of a particular solvent is made according to the particular drug and other substances used in a particular formulation, which is intended to entrap a particular active agent and which is intended to be released at a particular intended rate and duration. In control determined according to the properties of the obtained solution

The use of different types of volatile organic solutions and the absence of water throughout the process enables the formation of a homogeneous water-resistant lipid-based matrix composition according to the present disclosure. According to various embodiments, the first and second solvents may be the same or different. According to some embodiments, one solvent may be non-polar and the other is preferably miscible with water.

In another embodiment, the matrix composition of the methods and compositions of the present invention is substantially anhydrous. In another embodiment, "substantially anhydrous" refers to a composition comprising less than 1% water by weight. In another embodiment, the term refers to a composition comprising less than 0.8% by weight of water. In another embodiment, the term refers to a composition comprising less than 0.6% by weight of water. In another embodiment, the term refers to a composition comprising less than 0.4% by weight of water. In another embodiment, the term refers to a composition comprising less than 0.2% by weight of water. In another embodiment, the term refers to the absence of an amount of water that affects the water resistance properties of the composition. In another embodiment, the term refers to a composition prepared without any aqueous solvent. In another embodiment, lipid saturation can be achieved using a substantially anhydrous process to produce a composition, as described herein. Lipid saturation gives the matrix composition the ability to resist substantial degradation in vivo; thus, the matrix composition exhibits the ability to modulate extended release over a range of days, weeks or months.

In another embodiment, the matrix composition is substantially anhydrous. By "substantially free" is meant a composition comprising less than 0.1% by weight of water. In another embodiment, the term refers to a composition comprising less than 0.08% by weight of water. In another embodiment, the term refers to a composition comprising less than 0.06% water by weight. In another embodiment, the term refers to a composition comprising less than 0.04% water by weight. In another embodiment, the term refers to a composition comprising less than 0.02% by weight of water. In another embodiment, the term refers to a composition comprising less than 0.01% water by weight. Each possibility represents a separate embodiment of the invention.

In another embodiment, the matrix composition is anhydrous. In another embodiment, the term refers to a composition that does not contain a detectable amount of water. Each possibility represents a separate embodiment of the invention.

In another embodiment, the present invention provides a method of producing a matrix composition, the method comprising the steps of: (a) combining the following with a non-polar volatile organic solvent: (i) a biodegradable polyester and (ii) a sterol; (b) combining the following with a volatile organic solvent that is miscible with water: (i) an active agent selected from the group consisting of: non-steroidal anti-inflammatory drugs (NSAIDs), antibiotics, antifungals, steroids, anticancer agents, and osteogenic factors and bone resorption inhibitors; (ii) phosphatidylethanolamine; and (iii) phosphatidylcholine; and (c) mixing and homogenizing the products produced in steps (a) and (b). In another embodiment, the phosphatidylethanolamine is contained in a non-polar volatile organic solvent, rather than a volatile organic solvent that is miscible with water. In another embodiment, the biodegradable polyester is selected from the group consisting of PLA, PGA, and PLGA. In another embodiment, the biodegradable polymer is any other suitable biodegradable polyester known in the art. In another embodiment, the mixture comprising the non-polar organic solvent is homogenized before mixing it with the mixture organic solvent. In another embodiment, the mixture comprising the water-miscible organic solvent is homogenized before mixing it with the mixture comprising the non-polar organic solvent. In another embodiment, the polymer in the mixture of step (a) is lipid saturated. In another embodiment, the matrix composition is lipid saturated (lipid saturated). Each possibility represents a separate embodiment of the invention.

In another embodiment, the matrix composition of the present invention may be used to completely or partially coat the surface of different substrates. In another embodiment, the substrate to be coated comprises at least one material selected from the group consisting of: carbon fiber, stainless steel, cobalt-chromium, titanium alloys, tantalum, ceramics, and collagen or gelatin. In another embodiment, the substrate may comprise any medical device, such as orthopedic nails, orthopedic screws, orthopedic staples (orthopedic staples), orthopedic wires and orthopedic pins (orthopedic pins) used in orthopedic surgery, metallic or polymeric implants used in both orthopedic surgery and periodontal surgery, bone filler particles (bone filler particles) and resorbable gelatin sponges. The bone filler particles may be any of allogeneic bone particles (i.e., from human sources), xenogeneic bone particles (i.e., from animal sources), and artificial bone particles. In another embodiment, treatment with a coated substrate and application of the coated substrate will follow procedures known in the art for treatment and application of similar uncoated substrates. In another embodiment, the bone filler particle coated with the biodegradable matrix of the present invention is administered substantially as a single component (not as part of a mixture with other components). Optionally, the coated bone filler particles are mixed with any other commercially available bone filler particles or autologous bone prior to administration. In another embodiment, the mixture of bone filler particles comprises at least one of: uncoated particles, particles coated with a matrix composition incorporating a pharmaceutically active agent, particles coated with a matrix composition incorporating a plurality of pharmaceutically active agents, or a combination thereof. In another embodiment, the amounts, ratios and types of ingredients forming the matrix composition of the present invention are varied to modulate the polymer-lipid basis (polymer-lipid basis) according to the biophysical/biochemical properties of the pharmaceutically active agent, the therapeutically effective dose of the pharmaceutically active agent and the desired sustained release time period (typically ranging from days to months).

It is emphasized that the sustained release period using the composition of the invention may be set taking into account two main factors: (i) the weight ratio between the polymer and the lipid content, in particular phospholipids having fatty acid moieties of at least 14 carbons, and (ii) the biochemical and/or biophysical properties of the biopolymer and the lipid. In particular, the degradation rate of the polymer and the fluidity of the lipid should be considered. For example, a PLGA (85:15) polymer will degrade more slowly than a PLGA (50:50) polymer. Phosphatidylcholine (14:0) is more mobile (less rigid and less ordered) at body temperature than phosphatidylcholine (18: 0). Thus, for example, the release rate of a drug incorporated into a matrix composition comprising PLGA (85:15) and phosphatidylcholine (18:0) will be slower than the release rate of a drug incorporated into a matrix comprising PLGA (50:50) and phosphatidylcholine (14: 0). Another aspect that will determine the release rate is the physical characteristics of the encapsulated or impregnated drug. In addition, the release rate of the drug can be further controlled by adding additional lipids to the formulation of the second solution. This may include fatty acids of varying lengths, such as lauric acid (C12:0), membrane active sterols (such as cholesterol) or other phospholipids, such as phosphatidylethanolamine. According to various embodiments, the active agent is released from the composition over a desired period of time ranging between days to months.

These and other features and advantages of the present invention will be more readily understood and appreciated from the following detailed description of the invention.

Brief Description of Drawings

FIG. 1(A and B): doxycycline hydrochloride (DOX) encapsulated in the formulation of the present invention is released continuously for at least 3 weeks. A) DOX released from a matrix comprising PLGA 85:15, cholesterol, alpha-tocopherol, and DSPC (18: 0); B) DOX released from a matrix comprising PLGA 85:15, cholesterol and DSPC (18: 0).

FIG. 2: most (70%) of the DOX encapsulated in the formulation of the invention is released according to zero order kinetics. The Y-axis represents DOX release rate in μ g/ml/hr.

FIG. 3: the macrostructure of the bone particles is not affected by the coating with the formulation of the invention. (A) The initial structure of the bone particles; (B) bone particles coated with a formulation of the invention encapsulating DOX; (C) bone particles of (B) after incubation in serum for 60 days.

FIG. 4: the matrix formulation coating the surface of the bone particles undergoes gradual surface degradation. (A) An untreated surface; (B) the surface of the coated bone particles; (C) the surface of the coated bone particles after 1 day in 10% FBS at 37 ℃; (D) the surface of the coated bone particles after 30 days in 10% FBS at 37 ℃; (E) surface of coated bone particles after 60 days in 10% FBS at 37 ℃.

FIG. 5: the ordered structure of the matrix formulations (PLGA (85:15), DPPC (16:0), cholesterol 10%) is shown by electron microscopy (. times.18,000). The bright color lines represent the polymer, while the lipids are represented by dark filling between the polymer materials.

FIG. 6: the particular polymer/lipid composition of the formulation of the invention determines the release rate of a given drug. Effect of different concentrations of Lauric Acid (LA) and Phosphatidylethanolamine (PE) given as w/w% of the formulation on the release time of 90% of the encapsulated drug.

FIG. 7: use of dimethyl phosphatidylethanolamine (DMPE) for cholesterol in the formulations of the present invention. The release profile of DOX from bone particles coated with a formulation comprising Dimethyldimyristoylphosphatidylethanolamine (DMPE) in a first organic solvent during preparation (diamonds) was compared to the release profile of bone particles coated with a formulation comprising cholesterol after hydration of the bone particles (37 ℃, 5% plasma) at the same preparation stage (squares).

FIG. 8: the nature of the polymer and the phospholipid determines the release rate of the encapsulated pharmaceutically active agent. Flurbiprofen is released from a matrix comprising PLGA (50:50) and DMPC (14:0), while doxycycline hydrochloride is released from a matrix composition comprising PLGA (85:15) and DSPC (18: 0).

FIG. 9: DOX release profile from bone particles coated with a formulation comprising DOX and mixed with similar uncoated bone particles at a 1: 4 ratio (diamonds) to free DOX mixed with the same amount of uncoated bone particles after hydration of the bone particles (37 ℃, 5% serum).

FIG. 10: the duration of drug release from bone particles coated with the formulation of the present invention is linearly dependent on formulation quality. Bone particles (12 mg/sample) were coated with different mass of formulations containing DOX (X-axis reflects formulation mass in mg). After hydration of the bone particles, the release of DOX from the formulation was monitored. The Y-axis reflects the day in which the cumulative release of entrapped DOX exceeds 90% of the total entrapped dose.

FIG. 11: antifungal agentsThiabenzimidazole(TBZ) release profile from bone particles coated with formulations (PLGA 50:50, cholesterol and DMPC (14:0)) comprising TBZ (10% of the total mass of the formulation).

FIG. 12: antibiotics were released from absorbable gelatin sponge (gelatamp. roeko) coated with the matrix formulations of the present invention (PLGA 75:25, PC16:0, cholesterol 10% and DOX 10%). The release of DOX from absorbable gelatin sponges pre-wetted with DOX solutions with similar drug doses was used as a control.

FIG. 13: degradation of the bone particle coating formulation is reflected by surface elemental analysis. After hydration of the coated bone particles, the percentages of carbon, calcium and phosphorus atoms on the surface of the coated bone particles were monitored by SEM. The X-axis represents the time after hydration of the coated bone sample.

FIG. 14: turbidity analysis of supernatant solution of hydrated bone particles (5% serum): 4 different types of bone particles were analyzed: (i) common uncoated bone particles (ii) bone particles coated with the matrix composition of the invention having DOX as pharmaceutically active agent (iii) bone particles coated with DPPC and DOX and (iv) bone particles coated with PLGA. Turbidity (a) of the supernatant in which the bone particles were immersed was measured 1 hour after hydration and at 37 ℃. The hydration medium was replaced with fresh medium after one hour incubation and turbidity (B) was measured after 23 hours incubation at 37 ℃. An electron microscope image (C) of a hydration solution in which the bone particles coated with the matrix formulation of the present invention were immersed was collected 24 hours after hydration at 37 ℃. Size distribution (D) and zeta potential (E) analysis of the material released from the hydrated bone particles.

FIG. 15: encapsulated DOX and fluorescently labeled phosphatidylcholine (NBD-PC) were co-released from the surface of the coated bone particles into the surrounding medium at zero order kinetics (5% FBS at 37 ℃).

FIG. 16: small angle X-ray scattering (SAXS) analysis of bone particles coated with the matrix formulations of the invention (PLGA 85:15, DPPC 16:0, DOX) revealed that the matrix had an ordered structure. As a control, the scattering curves of dried DOPS (18:1) powder and normal uncoated bone particles were recorded.

FIG. 17: A. differential Scanning Calorimetry (DSC) showed that cholesterol reduced the caloric uptake of PLGA upon heating. B. The reduction in PLGA caloric uptake is evident in the presence of other lipids such as the antioxidant alpha-tocopherol, but not in the presence of lipids such as mineral oil (containing hydrocarbons with carbon chains of C12-C18).

FIG. 18: dental metal implants made with titanium coated with a matrix formulation comprising PLGA (18:15), DSPC (18:0), cholesterol 10% and 10% DOX. A. Uncoated dental implant b. coated implant. The bright color of the coated implant under UV light is due to the fluorescent emission of DOX.

Detailed description of the invention

The term "controlled release" refers to control of the rate and/or amount of pharmaceutically active agent delivered by the matrix composition of the present invention. The controlled release may be continuous or discontinuous, and/or linear or nonlinear.

The term "sustained release" means that the pharmaceutically active agent is released over an extended period of time.

In certain embodiments, the present invention provides a matrix composition comprising: (a) a biodegradable polyester; (b) phosphoglycerides having a hydrocarbon moiety of at least 14 carbons; and (c) a pharmaceutically active agent. According to some embodiments, the pharmaceutical agent is selected from the group consisting of: antibiotics, antifungal agents, NSAIDs, steroids, anticancer agents, osteogenic factors, and bone resorption inhibitors.

In certain embodiments, the phosphoglyceride is a phospholipid. In some embodiments, the phospholipid is a phosphatidylcholine having fatty acid moieties of at least 14 carbons. In another embodiment, the composition further comprises phosphatidylethanolamine having a fatty acid moiety of at least 14 carbons. In another embodiment, the composition further comprises a sterol. In some embodiments, the sterol is cholesterol.

In another embodiment, the matrix composition is lipid saturated. As used herein, "lipid saturation" refers to the polymer of the matrix composition being saturated with lipids (including phospholipids) and any other lipids that may be present that bind any hydrophobic drug and targeting moiety present in the matrix. The matrix composition is saturated with any lipids present. The lipid-saturated matrix of the present invention exhibits the additional advantage of not requiring synthetic emulsifiers or surfactants, such as polyvinyl alcohol; thus, the compositions of the present invention are generally substantially free of polyvinyl alcohol. Polymers used to determine the achievement of lipid saturation: the method of lipid ratio and the method of determining the lipid saturation of the matrix are described below.

In another embodiment, the matrix composition is homogeneous. In another embodiment, the matrix composition is in the form of a lipid-saturated matrix, the shape and boundaries of which are defined by the biodegradable polymer. In another embodiment, the matrix composition is in the form of an implant. Preferably, the polyester, phosphatidylethanolamine, and sterol are incorporated into the matrix composition. In another embodiment, phosphatidylcholine is also incorporated into the matrix composition. In another embodiment, an antibiotic is also incorporated into the matrix composition. In another embodiment, the antibiotic has low water solubility. In another embodiment, the antibiotic is a hydrophobic antibiotic. In another embodiment, the antibiotic is an amphiphilic antibiotic. In another embodiment, the composition further comprises a non-steroidal anti-inflammatory drug (NSAID). In another embodiment, the NSAID is also incorporated into the matrix composition. In another embodiment, the NSAID has low water solubility. Each possibility represents a separate embodiment of the invention.

In one embodiment, the present invention provides a matrix composition comprising: (a) a biodegradable polyester; (b) a sterol; (c) phosphatidylethanolamine having a fatty acid moiety of at least 14 carbons; (d) a phosphatidylcholine having fatty acid moieties of at least 14 carbons; and (e) an antibiotic or antifungal agent. In another embodiment, the matrix composition is lipid saturated. Preferably, the polyester, phosphatidylethanolamine, and sterol are incorporated into the matrix composition. In another embodiment, phosphatidylcholine is also incorporated into the matrix composition. In another embodiment, an antibiotic is also incorporated into the matrix composition. In another embodiment, the antibiotic has low water solubility. In another embodiment, the antibiotic is a hydrophobic antibiotic. In another embodiment, the antibiotic is an amphiphilic antibiotic. In another embodiment, the composition further comprises a non-steroidal anti-inflammatory drug (NSAID). In another embodiment, the NSAID is also incorporated into the matrix composition. In another embodiment, the NSAID has low water solubility. In another embodiment, the matrix composition is in the form of a lipid-saturated matrix, the shape and boundaries of which are influenced by the properties of the biodegradable polymer. In another embodiment, the matrix composition is in the form of an implant. In another embodiment, the matrix composition is homogeneous. Each possibility represents a separate embodiment of the invention.

In another embodiment, the present invention provides a matrix composition comprising: (a) a biodegradable polyester; (b) a sterol; (c) phosphatidylethanolamine having a fatty acid moiety of at least 14 carbons; (d) a phosphatidylcholine having fatty acid moieties of at least 14 carbons; (e) non-steroidal anti-inflammatory drugs (NSAIDs). In another embodiment, the matrix composition is lipid saturated. Preferably, the polyester, phosphatidylethanolamine, and sterol are incorporated into the matrix composition. In another embodiment, phosphatidylcholine is also incorporated into the matrix composition. In another embodiment, the NSAID is also incorporated into the matrix composition. In another embodiment, the NSAID has low water solubility. In another embodiment, the NSAID is a hydrophobic NSAID. In another embodiment, the NSAID is an amphiphilic NSAID. In another embodiment, the matrix composition is in the form of a lipid-saturated matrix, the shape and boundaries of which are defined by the biodegradable polymer. In another embodiment, the matrix composition is in the form of an implant. In another embodiment, the matrix composition is homogeneous. Each possibility represents a separate embodiment of the invention.

In another embodiment, the present invention provides a matrix composition comprising: (a) a biodegradable polyester; (b) a sterol; (c) phosphatidylethanolamine having a fatty acid moiety having at least 14 carbons; (d) a phosphatidylcholine having fatty acid moieties of at least 14 carbons; and (e) an osteogenic factor or a bone resorption inhibitor. In another embodiment, the matrix composition is lipid saturated. Preferably, the polyester, phosphatidylethanolamine, and sterol are incorporated into the matrix composition. In another embodiment, phosphatidylcholine is also incorporated into the matrix composition. In another embodiment, a bone resorption inhibitor is also incorporated into the matrix composition. In another embodiment, the bone resorption inhibitor has low water solubility. In another embodiment, the bone resorption inhibitor is a hydrophobic bone resorption inhibitor. In another embodiment, the bone resorption inhibitor is an amphiphilic bone resorption inhibitor. In another embodiment, the composition further comprises an NSAID. In another embodiment, the NSAID is also incorporated into the matrix composition. In another embodiment, the matrix composition is in the form of a lipid-saturated matrix, the shape and boundaries of which are defined by the biodegradable polymer. In another embodiment, the matrix composition is in the form of an implant. In another embodiment, the matrix composition is homogeneous. Each possibility represents a separate embodiment of the invention.

In another embodiment, the present invention provides a matrix composition comprising: (a) a biodegradable polyester; (b) a sterol; (c) phosphatidylethanolamine having a fatty acid moiety of at least 14 carbons; (d) a phosphatidylcholine having fatty acid moieties of at least 14 carbons; (e) an active agent; and (f) a targeting moiety capable of interacting with a surface molecule, target molecule or target surface of the target cell. In another embodiment, the matrix composition is lipid saturated. In another embodiment, the active agent is selected from the group consisting of: NSAIDs, antibiotics and bone resorption inhibitors. In another embodiment, the polymer and phospholipid form a substantially anhydrous matrix composition. In another embodiment, the active agent and targeting moiety are incorporated into a lipid vesicle. In another embodiment, the matrix composition is in the form of a lipid-saturated matrix, the shape and boundaries of which are defined by the biodegradable polymer. In another embodiment, the matrix composition is in the form of an implant. In another embodiment, the matrix composition is homogeneous. Each possibility represents a separate embodiment of the invention.

In another embodiment, the biodegradable polyester of the methods and compositions of the present invention is associated with the sterol via hydrogen bonding.

As provided herein, the matrix composition of the methods and compositions of the present invention can be molded into three-dimensional configurations having different thicknesses and shapes. Thus, the formed matrix may be produced to obtain specific shapes, including spheres, cubes, rods, tubes, sheets or wires. In the case of lyophilization, the shape is determined by the shape of the mold or support, which may be made of any inert material and may be in contact with the substrate on all sides for a sphere or cube, or on a limited number of sides for a sheet. The matrix may be shaped in the form of a body cavity, as required by the implant design. Removing portions of the matrix with scissors, a scalpel, a laser beam, or any other cutting tool can produce any fine machining required in the three-dimensional structure. Each possibility represents a separate embodiment of the invention.

Advantageously, the matrix composition of the present invention is prepared by a process that does not involve the formation of an emulsion and that can avoid the use of an aqueous medium together. The production of a substantially dry emulsion necessarily produces vesicles or microspheres. Rather, rather than forming a homogeneous liquid mixture from a matrix produced by an aqueous medium, the homogeneous liquid mixture can be shaped or formed into a three-dimensional article having any shape or can coat the surface of different substrates. To produce shaped or coated articles, a mixture of polymer and lipid and active ingredient in a suitable selected volatile organic solvent will be used to coat the desired surface or to conform to the desired shape.

The matrix composition of the methods and compositions of the present invention is capable of coating the surface of various substrates. The substrate to be coated comprises a material selected from the group consisting of: carbon fiber, stainless steel, cobalt-chromium, titanium alloys, tantalum, ceramics, and collagen or gelatin. In particular, the substrate may include any medical device, such as orthopedic nails, orthopedic screws, orthopedic staples, orthopedic wires, and orthopedic needles used in orthopedic surgery, metallic or polymeric implants used in orthopedic surgery and periodontal surgery, bone filler particles, and resorbable gelatin sponges. The bone filler particles may be selected from any of allogeneic bone particles (i.e., from human origin), xenogeneic bone particles (i.e., from animal origin), and artificial bone particles.

According to some embodiments, the matrix composition of the present invention is used as a bone graft material. This term refers to natural or synthetic materials that support the attachment of new osteoblasts and osteoprogenitors or can induce differentiation of undifferentiated stem cells or osteoprogenitors into osteoblasts. In another embodiment, the bone graft material is selected from the group consisting of: allografts, allografts (alloplasts), xenografts, and autobone grafts. In other embodiments, the lipid matrix of the present invention may also be used in conjunction with collagen membranes or collagen sponges or gelatin sponges or the like.

Lipid

A "phospholipid" is a phosphoglyceride having a single phosphatidyl bond (linkage) on the glycerol backbone and fatty acids at the remaining two positions. However, it is expressly understood that phosphoglycerides having hydrocarbon chains of at least 14 carbons other than the fatty acid residue, including alkyl chains, alkenyl chains, or any other hydrocarbon chains, are included within the scope of the present invention. The linkage may be an ether linkage instead of an acyl linkage in the phospholipid.

"Phosphatidylcholine" refers to a phosphoglyceride having a phosphorylcholine head group. In another embodiment, the phosphatidylcholine compound has the following structure:

r and R¹The moiety is a fatty acid, typically a naturally occurring fatty acid or a derivative of a naturally occurring fatty acid. In some embodiments, the fatty acid moiety is a saturated fatty acid moiety. In some embodiments, the fatty acid moiety is an unsaturated fatty acid moiety. "saturated" means that no double bonds are present in the hydrocarbon chain. In another embodiment, the fatty acid moiety has at least 14 carbon atoms. In another embodiment, the fatty acid moiety has 16 carbon atoms. In another embodiment, the fatty acid moiety has 18 carbon atoms. In another embodiment, the fatty acid moiety has 16 to 18 carbon atoms. In another embodiment, the fatty acid moieties are selected such that the resulting gel to liquid crystal transition temperature of the matrix is at least 40 ℃. In another embodiment, the fatty acid moieties are both palmitoyl. In another embodiment, the fatty acid moieties are both stearoyl. In another embodiment, the fatty acid moieties are all arachidoyl (arachidoyl). In another embodiment, the fatty acid moieties are palmitoyl and stearoyl. In another embodiment, the fatty acid moieties are palmitoyl and arachidoyl. In another embodiment, the fatty acidSome are arachidoyl and stearoyl. Each possibility represents a separate embodiment of the invention.

In another embodiment, the phosphatidylcholine is a naturally occurring phosphatidylcholine. In another embodiment, the phosphatidylcholine is synthetic phosphatidylcholine. In another embodiment, the phosphatidylcholine contains a naturally occurring isotopic distribution. In another embodiment, the phosphatidylcholine is deuterated phosphatidylcholine. In another embodiment, the phosphatidylcholine is labeled with any other isotope or label. Preferably, the phosphatidylcholine is symmetric phosphatidylcholine (i.e., phosphatidylcholine wherein both fatty acid moieties are the same). In another embodiment, the phosphatidylcholine is asymmetric phosphatidylcholine.

Non-limiting examples of phosphatidylcholines are 1, 2-distearoyl-sn-glycero-3-phosphocholine (DSPC), dioleoyl-phosphatidylcholine (DOPC), 1-palmitoyl-2-oleoyl-phosphatidylcholine, and phosphatidylcholine modified with any of the fatty acid moieties listed above. In another embodiment, the phosphatidylcholine is selected from the group consisting of DSPC and DOPC and 1-palmitoyl-2-oleoyl-phosphatidylcholine.

In another embodiment, the phosphatidylcholine is any other phosphatidylcholine known in the art. Each phosphatidylcholine represents a separate embodiment of the present invention.

"phosphatidylethanolamine" refers to a phosphoglyceride having a phosphoethanolamine headgroup. In another embodiment, the phosphatidylethanolamine compound has the following structure:

R₁and R₂The moiety is a fatty acid, typically a naturally occurring fatty acid or a derivative of a naturally occurring fatty acid. In another embodiment, fatty acid moietiesIs a saturated fatty acid moiety. In another embodiment, "saturated" means that no double bonds are present in the hydrocarbon chain. In another embodiment, the fatty acid moiety has at least 14 carbon atoms. In another embodiment, the fatty acid moiety has at least 16 carbon atoms. In another embodiment, the fatty acid moiety has 14 carbon atoms. In another embodiment, the fatty acid moiety has 16 carbon atoms. In another embodiment, the fatty acid moiety has 18 carbon atoms. In another embodiment, the fatty acid moiety has 14 to 18 carbon atoms. In another embodiment, the fatty acid moiety has 14 to 16 carbon atoms. In another embodiment, the fatty acid moiety has 16 to 18 carbon atoms. In another embodiment, the fatty acid moieties are selected such that the resulting gel to liquid crystal transition temperature of the matrix is at least 40 ℃. In another embodiment, the fatty acid moieties are both myristoyl. In another embodiment, the fatty acid moieties are both palmitoyl. In another embodiment, the fatty acid moieties are both stearoyl. In another embodiment, the fatty acid moieties are both arachidoyl. In another embodiment, the fatty acid moieties are myristoyl and stearoyl. In another embodiment, the fatty acid moieties are myristoyl and arachidoyl. In another embodiment, the fatty acid moieties are myristoyl and palmitoyl. In another embodiment, the fatty acid moieties are palmitoyl and stearoyl. In another embodiment, the fatty acid moieties are palmitoyl and arachidoyl. In another embodiment, the fatty acid moieties are arachidoyl and stearoyl. Each possibility represents a separate embodiment of the invention.

In another embodiment, the phosphatidylethanolamine is a naturally occurring phosphatidylethanolamine. In another embodiment, the phosphatidylethanolamine is a synthetic phosphatidylethanolamine. In another embodiment, the phosphatidylethanolamine is a deuterated phosphatidylethanolamine. In another embodiment, the phosphatidylethanolamine is labeled with any other isotope or label. In another embodiment, the phosphatidylethanolamine comprises a naturally occurring isotopic distribution. Preferably, the phosphatidylethanolamine is symmetric phosphatidylethanolamine. In another embodiment, the phosphatidylethanolamine is an asymmetric phosphatidylethanolamine.

Non-limiting examples of phosphatidylethanolamines are Dimethyldimyristoylphosphatidylethanolamine (DMPE) and Dipalmitoylphosphatidylethanolamine (DPPE), and phosphatidylethanolamines modified with any of the fatty acid moieties listed above. In another embodiment, the phosphatidylethanolamine is selected from the group consisting of DMPE and DPPE.

In another embodiment, the phosphatidylethanolamine is any other phosphatidylethanolamine known in the art. Each phosphatidylethanolamine represents a separate embodiment of the present invention.

In one embodiment, "sterol" refers to steroids having a hydroxyl group at the 3-position of the A ring. In another embodiment, the term refers to steroids having the structure:

in another embodiment, the sterols of the methods and compositions of the invention are substantially animal sterols. In another embodiment, the sterol is cholesterol:

in another embodiment, the sterol is any other animal sterol known in the art. In another embodiment, the moles of sterols are up to 40% of the moles of total lipid present. In another embodiment, the sterol is incorporated into the matrix composition. Each possibility represents a separate embodiment of the invention.

In another embodiment, the cholesterol is present in an amount of 10 to 50 percent by total weight of the lipid content of the matrix composition. In another embodiment, the weight percentage is 20-50%. In another embodiment, the weight percentage is 10-40%. In another embodiment, the weight percentage is 30-50%. In another embodiment, the weight percentage is 20-60%. In another embodiment, the weight percentage is 25-55%. In another embodiment, the weight percentage is 35-55%. In another embodiment, the weight percentage is 30-60%. In another embodiment, the weight percentage is 30-55%. In another embodiment, the weight percentage is 20-50%. In another embodiment, the weight percentage is 25-55%. Each possibility represents a separate embodiment of the invention.

In another embodiment, the composition of the present invention further comprises a lipid other than phosphatidylcholine, phosphatidylethanolamine or sterol. In another embodiment, the additional lipid is a phosphoglyceride. In another embodiment, the additional lipid is selected from the group consisting of phosphatidylserine, phosphatidylglycerol and phosphatidylinositol. In another embodiment, the additional lipid is selected from the group consisting of phosphatidylserine, phosphatidylglycerol, phosphatidylinositol, and sphingomyelin. In another embodiment, there is a combination of any 2 or more of the above additional lipids. In another embodiment, the polymer, phosphatidylcholine, phosphatidylethanolamine, sterol, and additional lipid are all incorporated into the matrix composition. Each possibility represents a separate embodiment of the invention.

In another embodiment, Phosphatidylcholine (PC) comprises at least 30% of the total lipid content of the matrix composition. In another embodiment, the PC comprises at least 35% of the total lipid content. In another embodiment, the PC comprises at least 40% of the total lipid content. In another embodiment, the PC comprises at least 45% of the total lipid content. In another embodiment, the PC comprises at least 50% of the total lipid content. In another embodiment, the PC comprises at least 55% of the total lipid content. In another embodiment, the PC comprises at least 60% of the total lipid content. In another embodiment, the PC comprises at least 65% of the total lipid content. In another embodiment, the PC comprises at least 70% of the total lipid content. In another embodiment, the PC comprises at least 75% of the total lipid content. In another embodiment, the PC comprises at least 80% of the total lipid content. In another embodiment, the PC comprises at least 85% of the total lipid content. In another embodiment, PC makes up at least 90% of the total lipid content. In another embodiment, the PC comprises at least 95% of the total lipid content. In another embodiment, the PC comprises more than 95% of the total lipid content. Each possibility represents a separate embodiment of the invention.

In another embodiment, the composition of the invention further comprises phosphatidylserine. "Phosphatidylserine" refers to a phosphoglyceride having a phosphoryl serine head group. In another embodiment, the phosphatidylserine compound has the following structure:

R₁and R₂The moiety is a fatty acid, typically a naturally occurring fatty acid or a derivative of a naturally occurring fatty acid. In another embodiment, the fatty acid moiety is a saturated fatty acid moiety. In another embodiment, the fatty acid moiety has at least 14 carbon atoms. In another embodiment, the fatty acid moiety has at least 16 carbon atoms. In another embodiment, the fatty acid moieties are selected such that the resulting gel to liquid crystal transition temperature of the matrix is at least 40 ℃. In another embodiment, the fatty acid moieties are both myristoyl. In another embodiment, the fatty acid moieties are all palmitoyl. In another embodiment, the fatty acid moieties are both stearoyl. In another embodiment, the fatty acid moieties are both arachidoyl. In another embodiment, the fatty acid moieties are myristoyl and stearoyl. In another embodiment, the fatty acid moiety is a combination of two of the above fatty acid moieties.

In another embodiment, the phosphatidylserine is a naturally occurring phosphatidylserine. In another embodiment, the phosphatidylserine is a synthetic phosphatidylserine. In another embodiment, the phosphatidylserine is a deuterated phosphatidylserine. In another embodiment, the phosphatidylserine is labeled with any other isotope or label. In another embodiment, the phosphatidylserine comprises a naturally occurring isotopic distribution. In another embodiment, the phosphatidylserine is a symmetric phosphatidylserine. In another embodiment, the phosphatidylserine is an asymmetric phosphatidylserine.

A non-limiting example of a phosphatidylserine is a phosphatidylserine modified with any of the fatty acid moieties listed above. In another embodiment, the phosphatidylserine is any other phosphatidylserine known in the art. Each phosphatidylserine represents a separate embodiment of the present invention.

In another embodiment, the composition of the invention further comprises phosphatidylglycerol. "phosphatidylglycerol" refers to a phosphoglyceride having a phosphoryl glycerol head group. In another embodiment, the phosphatidylglycerol compound has the following structure:

the 2 linkages on the left are linked to fatty acids, typically naturally occurring fatty acids or derivatives of naturally occurring fatty acids. In another embodiment, the phosphatidylglycerol is a naturally occurring phosphatidylglycerol. In another embodiment, the phosphatidylglycerol is a synthetic phosphatidylglycerol. In another embodiment, the phosphatidylglycerol is a deuterated phosphatidylglycerol. In another embodiment, the phosphatidylglycerol is labeled with any other isotope or label. In another embodiment, the phosphatidylglycerol comprises a naturally occurring isotopic distribution. In another embodiment, the phosphatidylglycerol is a symmetric phosphatidylglycerol. In another embodiment, the phosphatidylglycerol is an asymmetric phosphatidylglycerol. In another embodiment, the term includes a diphosphatidylglycerol compound having the structure:

the R and R' moieties are fatty acids, typically naturally occurring fatty acids or derivatives of naturally occurring fatty acids. In another embodiment, the fatty acid moiety is a saturated fatty acid moiety. In another embodiment, the fatty acid moiety has at least 14 carbon atoms. In another embodiment, the fatty acid moiety has at least 16 carbon atoms. In another embodiment, the fatty acid moieties are selected such that the resulting gel to liquid crystal transition temperature of the matrix is at least 40 ℃. In another embodiment, the fatty acid moieties are both myristoyl. In another embodiment, the fatty acid moieties are both palmitoyl. In another embodiment, the fatty acid moieties are both stearoyl. In another embodiment, the fatty acid moieties are both arachidoyl. In another embodiment, the fatty acid moieties are myristoyl and stearoyl. In another embodiment, the fatty acid moiety is a combination of two of the above fatty acid moieties.

A non-limiting example of a phosphatidylglycerol is a phosphatidylglycerol modified with any of the fatty acid moieties listed above. In another embodiment, the phosphatidylglycerol is any other phosphatidylglycerol known in the art. Each phosphatidylglycerol represents a separate embodiment of the present invention.

In another embodiment, the composition of the invention further comprises phosphatidylinositol. "phosphatidylinositol" refers to a phosphoglyceride having a phosphatidylinositol headgroup. In another embodiment, the phosphatidylinositol compound has the following structure:

R₁and R₂The moiety is a fatty acid, typically a naturally occurring fatty acid or a derivative of a naturally occurring fatty acid. In another embodiment, the fatty acid moiety is a saturated fatty acid moiety. In another embodiment, the fatty acid moiety has at least 14 carbon atoms. In another embodiment, the fatty acid moiety has at least 16 carbon atoms. In another embodiment, the fatty acid moieties are selected such that the resulting gel to liquid crystal transition temperature of the matrix is at least 40 ℃. In another embodiment, the fatty acid moieties are both myristoyl. In another embodiment, the fatty acid moieties are both palmitoyl. In another embodiment, the fatty acid moieties are both stearoyl. In another embodiment, the fatty acid moieties are both arachidoyl. In another embodiment, the fatty acid moieties are myristoyl and stearoyl. In another embodiment, the fatty acid moiety is a combination of two of the above fatty acid moieties.

In another embodiment, the phosphatidylinositol is a naturally occurring phosphatidylinositol. In another embodiment, the phosphatidylinositol is synthetic phosphatidylinositol. In another embodiment, the phosphatidylinositol is a deuterated phosphatidylinositol. In another embodiment, the phosphatidylinositol is labeled with any other isotope or label. In another embodiment, the phosphatidylinositol comprises a naturally occurring isotopic distribution. In another embodiment, the phosphatidylinositol is symmetric phosphatidylinositol. In another embodiment, the phosphatidylinositol is an asymmetric phosphatidylinositol.

A non-limiting example of phosphatidylinositol is phosphatidylinositol modified by any of the fatty acid moieties listed above. In another embodiment, the phosphatidylinositol is any other phosphatidylinositol known in the art. Each phosphatidylinositol represents a separate embodiment of the present invention.

In another embodiment, the composition of the invention further comprises a sphingolipid. In another embodiment, the sphingolipid is a ceramide. In another embodiment, the sphingolipid is sphingomyelin. "sphingomyelin" refers to sphingosine-derived phospholipids. In another embodiment, the sphingomyelin compound has the following structure:

the R moiety is a fatty acid, typically a naturally occurring fatty acid or a derivative of a naturally occurring fatty acid. In another embodiment, the sphingomyelin is a naturally occurring sphingomyelin. In another embodiment, the sphingomyelin is synthetic sphingomyelin. In another embodiment, the sphingomyelin is a deuterated sphingomyelin. In another embodiment, the sphingomyelin is labeled with any other isotope or label. In another embodiment, the sphingomyelin comprises a naturally occurring isotopic distribution.

In another embodiment, the fatty acid moiety of the sphingomyelin of the methods and compositions of the invention has at least 14 carbon atoms. In another embodiment, the fatty acid moiety has at least 16 carbon atoms. In another embodiment, the fatty acid moieties are selected such that the resulting gel to liquid crystal transition temperature of the matrix is at least 40 ℃.

A non-limiting example of a sphingomyelin is a sphingomyelin modified by any of the fatty acid moieties listed above. In another embodiment, the sphingomyelin is any other sphingomyelin known in the art. Each sphingomyelin represents a separate embodiment of the invention.

"ceramide" refers to a compound having the structure:

the R moiety is a fatty acid, typically a naturally occurring fatty acid or a derivative of a naturally occurring fatty acid. In another embodiment, the fatty acid is a longer chain (to C)₂₄Or larger). In another embodiment, the fatty acid is a saturated fatty acid. In another embodiment, the fatty acid is a mono-olefinic fatty acid. In another embodiment, the fatty acid is an n-9 monoalkene fatty acid. In another embodiment, the fatty acid comprises a hydroxyl group at the 2-position. In another embodiment, the fatty acid is other suitable fatty acids known in the art. In another embodiment, the ceramide is a naturally occurring ceramide. In another embodiment, the ceramide is a synthetic ceramide. In another embodiment, the ceramide is incorporated into a matrix composition. Each possibility represents a separate embodiment of the invention.

Each sphingolipid represents a separate embodiment of the present invention.

In another embodiment, the composition of the invention further comprises a pegylated lipid (pegylated lipid). In another embodiment, the PEG moiety has a MW of 500-. In another embodiment, the PEG moiety has any other suitable MW. Non-limiting examples of suitable PEG-modified lipids include PEG moieties having methoxy end groups, such as PEG-modified phosphatidylethanolamine and phosphatidic acid (structures a and B), PEG-modified diacylglycerol and dihydrocarbyl glycerol (structures C and D), PEG-modified dihydrocarbylamine (structure E) and PEG-modified 1, 2-diacyloxypropyl-3-amine (structure F), as described below. In another embodiment, the PEG moiety has any other terminal group used in the art. In another embodiment, the pegylated lipid is selected from the group consisting of: PEG-modified phosphatidylethanolamine, PEG-modified phosphatidic acid, PEG-modified diacylglycerol, PEG-modified dihydrocarbylglycerol, PEG-modified dihydrocarbylamine, and PEG-modified 1, 2-diacyloxypropyl-3-amine. In another embodiment, the pegylated lipid is any other pegylated phospholipid known in the art. Each possibility represents a separate embodiment of the invention.

Preferably, the pegylated lipid is present in an amount of less than 10 mole percent of the total lipid in the matrix composition. In another embodiment, the percentage is less than 9 mole percent of total lipid. In another embodiment, the percentage is less than 8 mole percent. In another embodiment, the percentage is less than 7 mole percent. In another embodiment, the percentage is less than 6 mole percent. In another embodiment, the percentage is less than 5 mole percent. In another embodiment, the percentage is less than 4 mole percent. In another embodiment, the percentage is less than 3 mole percent. In another embodiment, the percentage is less than 2 mole percent. In another embodiment, the percentage is less than 1 mole percent. Each possibility represents a separate embodiment of the invention.

Polymer and method of making same

In another embodiment, the biodegradable polyester of the methods and compositions of the present invention is PLA (polylactic acid). "PLA" refers to poly (L-lactide), poly (D-lactide), and poly (DL-lactide). Representative structures of poly (DL-lactide) are described below:

in another embodiment, the polymer is PGA (polyglycolic acid). In another embodiment, the polymer is PLGA (poly (lactic-co-glycolic acid)). The PLA comprised in the PLGA may be any PLA known in the art, such as an enantiomeric or racemic mixture. Representative structures of PLGA are described below:

in another embodiment, the PLGA of the methods and compositions of the present invention has a lactic acid to glycolic acid ratio of 1: 1. In another embodiment, the ratio is 60: 40. In another embodiment, the ratio is 70: 30. In another embodiment, the ratio is 80: 20. In another embodiment, the ratio is 90: 10. In another embodiment, the ratio is 95: 5. In another embodiment, the ratio is another ratio suitable for use in an extended in vivo release profile, as defined herein. In another embodiment, the ratio is 50: 50. PLGA may be a random copolymer or a block copolymer. Each possibility represents a separate embodiment of the invention.

In another embodiment, the biodegradable polyester is selected from the group consisting of: polycaprolactone, polyhydroxyalkanoates, polypropylene fumarates (polypropylenefumarate), polyorthoesters, polyanhydrides, and polyalkylcyanoacrylates (polyalkylcyanoacrylates), with the proviso that the polyester contains a hydrogen bond acceptor moiety. In another embodiment, the biodegradable polyester is a block copolymer comprising a combination of any two monomers selected from the group consisting of: PLA, PGA, PLGA, polycaprolactone, polyhydroxyalkanoates, polypropylene fumarates, polyorthoesters, polyanhydrides, and polyalkylcyanoacrylates. In another embodiment, the biodegradable polyester is a random copolymer comprising a combination of any two of the monomers listed above. Each possibility represents a separate embodiment of the invention.

In another embodiment, the biodegradable polyesters of the methods and compositions of the present invention have a Molecular Weight (MW) of between about 10-40kDa (kilodaltons). In another embodiment, the MW is between about 5-50 KDa. In another embodiment, the MW is between about 15-40 KDa. In another embodiment, the MW is between about 20-40 KDa. In another embodiment, the MW is between about 15-35 KDa. In another embodiment, the MW is between about 10-35 KDa. In another embodiment, the MW is between about 10-30 KDa. In another embodiment, a mixture of PLGA polymers of different MW is used. In another embodiment, the different polymers all have a MW in one of the above ranges. Each possibility represents a separate embodiment of the invention.

Antibiotic

In another embodiment, the antibiotic of the methods and compositions of the present invention is doxycycline. In another embodiment, the antibiotic is a hydrophobic tetracycline. Non-limiting examples of hydrophobic tetracyclines are 6-demethyl-6-deoxytetracycline, 6-methylenetetracycline, minocycline (also known as 7-dimethylamino-6-demethyl-6-deoxytetracycline), and 13-phenylmercapto-a-6-deoxytetracycline. In another embodiment, the antibiotic is selected from the group consisting of doxycycline, tetracycline, and minocycline. In another embodiment, an antibiotic is incorporated into the matrix composition.

In another embodiment, the antibiotic is selected from the group consisting of: amoxicillin, amoxicillin/clavulanic acid, penicillin, metronidazole, clindamycin, chlortetracycline, demeclocycline, oxytetracycline, amikacin, noradriamycin, kanamycin, neomycin, netilmicin, streptomycin, tobramycin, cefadroxil, cefazolin, cephalexin, cephalothin, cefapirin, cefradine, cefaclor, cefamandole, cefmetazole, cefonicid, cefotetan, cefoxitin, cefpodoxime, cefprozil, cefuroxime, cefdinir, cefixime, cefoperazone, cefotaxime, ceftazidime, ceftibuten, cefazolin, ceftriaxone, cefepime, azithromycin, clarithromycin, dirithromycin, erythromycin, lincomycin, oleandomycin, baamicin, carbenicillin, cloxacillin, dicloxacillin, methicillin, mezlocillin, doxycycline, kanamycin, oxytetracycline, neomycin, cefixime, cefix, Nafcillin, oxacillin, piperacillin, ticarcillin, cinoxacin, ciprofloxacin, enoxacin, grepafloxacin, levofloxacin, lomefloxacin, nalidixic acid, norfloxacin, ofloxacin, sparfloxacin, sulfisoxazole, sulfacetamide, sulfadiazine, sulfamethoxazole, sulfisoxazole, dapsone, aztreonam, bacitracin, capreomycin, chloramphenicol, clofazimine, colistin sodium, polymyxin, cycloserine, fosfomycin, furazolidone, urotropin, nitrofurantoin, pentamidine, rifabutin, rifampin, spectinomycin, trimethoprim, trimetrexate and vancomycin.

In another embodiment, the bioactive ingredient is an antibacterial, such as chlorhexidine.

Each antibiotic represents a separate embodiment of the present invention.

NSAID

Any suitable NSAID may be incorporated into the matrix composition for sustained and/or controlled release. In one embodiment, the NSAID of the methods and compositions of the invention is flurbiprofen. In another embodiment, the NSAID is selected from the group consisting of ibuprofen and flurbiprofen. In another embodiment, the NSAID is selected from the group consisting of: ibuprofen, flurbiprofen, sodium aminosalicylate, choline magnesium trisalicylate, choline salicylate, diclofenac, diflunisal, etodolac, fenoprofen, indomethacin, ketoprofen, ketoconazole tromethamine, magnesium salicylate, meclofenamate, mefenamic acid, nabumetone, naproxen, oxaprozin, oxybutyzone, piroxicam, salsalate, sulindac, tolmetin.

Each NSAID represents a separate embodiment of the invention.

Steroids

In another embodiment, the active agent of the methods and compositions of the present invention is a steroid. According to one embodiment, the steroid is a steroidal anti-inflammatory drug. Non-limiting examples of steroidal anti-inflammatory drugs (SAIDs) used in the formulations of the present invention include, but are not limited to, corticosteroids such as: betamethasone, betamethasone valerate, cortisone, dexamethasone, 21-dexamethasone phosphate, fludrocortisone, dexamethasone, fluocinonide acetate, fludrolone acetonide acetate, naltrexone, fluocortolone, halcinonide, haloprednisone, hydrocortisone 17-valerate, hydrocortisone 17-butyrate, hydrocortisone 21-acetate, methylprednisolone, prednisolone 21-phosphate, prednisone, triamcinolone acetonide, cortolone, fluoroacetamide, fludrocortisone, diflorasone acetate, fluocinolone acetonide, medrysone, amfenap, benzoin, Benzonate, betamethasone and other esters thereof, predrysone, clocortolone, desinselone, desonide, dichlorosone, difluprednate, diflucortolone, flunisolone, fluocortolone, fluocinolone acetonide, Fluoromethalone, fluridone, flupredone, methylprednisolone, paramethasone, cortisone, hydrocortisone cypionate, cortodxolone, fluocinolone acetonide, fludrocortisone, fluocinonide ketal, medroxsone, amcinol, Benzonate, betamethasone benzoate, closterone acetate, clocortolone, desicatide, desoximetasone, dichloroacetate, difluprednate, fluocinolone, diflucortolone, diflunisal pivalate, fluocinonide, meflolone acetate, flupredlone, paramethasone acetate, prednisolone, prednate valerate, triamcinolone caproate, cortazole, formocortaol, and nilvakol.

Anticancer agent

As referred to herein, the term "anti-cancer agent" refers to any type of agent that can be used to treat cancer and/or cancer-related disorders. The anti-cancer agent can include any naturally occurring or synthetically produced molecule capable of directly or indirectly affecting the growth and/or viability of cancer cells, cancer tumors, and/or cancer-related disorders and symptoms. Anti-cancer agents may include, for example, naturally occurring proteins or peptides, modified proteins or peptides, recombinant proteins, chemically synthesized proteins or peptides, low oral bioavailability proteins or peptides, chemical molecules, synthetic chemical molecules, chemotherapeutic drugs, biologic therapeutic drugs, and the like, or any combination thereof. The anti-cancer agent can be cytotoxic (toxic to cells) and/or cytostatic (cytostatic) and/or antiproliferative to cancer cells and can exert its effect on cancer cells directly and/or indirectly. According to some embodiments, the anti-cancer agents may be administered alone or in combination and/or before and/or after one or more additional cancer treatments. Additional cancer treatments may include such treatments, such as, but not limited to: chemotherapy (using drugs to affect cancer cells), radiation therapy (using high energy radiation of various origins to affect cancer cells), biological therapy (therapy to help immune cells fight cancer), surgical procedures (surgical removal of cancerous tumors), gene therapy, bone marrow transplantation, any other therapy known in the art, or any combination thereof.

Non-limiting examples of anti-cancer agents and chemotherapeutic drugs may include such drugs as, but not limited to: alkaloids such as, but not limited to: docetaxel, etoposide, irinotecan, paclitaxel, teniposide, topotecan, vinblastine, vincristine, vindesine; alkylating agents such as, but not limited to: busulfan, improsulfan, piposulfan, benzotepa, carboquone, metotepipa, uretepa, altretamine, tritylamine, triethylphosphoramide, thiotepa, chlorambucil, cyclophosphamide, estramustine, ifosfamide, mechlorethamine, melphalan, novemebic, perfosfamide benzene mustard cholesterol, prednimustine, trofosfamide, uramustine, carmustine, chlorourethan, fotemustine, lomustine, nimustine, semustine, ramustine, dacarbazine, mannomustine, dibromomannitol, dibromodulcitol, pipobroman, temozolomide; antibiotics and analogs such as, but not limited to: doxorubicin, actinomycin, ampomycin, azaserine, bleomycin, actinomycin C, carubicin, chromamycin (Cromomycin), dactinomycin, daunorubicin, 6-diazo-5-oxo-L-norleucine, doxorubicin, epirubicin, idarubicin, minorit, mitomycin, mycophenolic acid, nogomycin (nogalamycin), olivomycin, pelubicin, pirarubicin, plicamycin, podomycin, puromycin, streptonigrin, streptozotocin, tubercidin, neat stastatin, zorubicin; antimetabolites such as, but not limited to: metformin, edatrexate, methotrexate, pirtrexin, pteropterin, raltitrexed, trimetrexate, cladribine (Cladridine), fludarabine, 6-mercaptopurine, pentostatin thioprimine (pentastatin Thiamipine), thioguanine, cyclocytidine, azacitidine, 6-azauride, carmofur, cytarabine, doxifluridine, etimefural, floxuridine, fluorouracil, gemcitabine, tegafur; platinum complexes such as, but not limited to: carboplatin, cisplatin, mepacrine, oxaliplatin; alkylating agents include, but are not limited to: busulfan (malilan, busulfan), chlorambucil (lechlene), ifosfamide (with or without MESNA), cyclophosphamide (cytoxan, neosar), prasfamide, melphalan, L-PAM (Ikelan), dacarbazine (DTIC-DOME), and temozolomide (temodar); anthracycline antibiotics include, but are not limited to: doxorubicin (Adriamycin, Doxil, Rubex), mitoxantrone (Novantrone), idarubicin (Idamycin), valrubicin (Valstar) and epirubicin (elence); antibiotics include, but are not limited to: dactinomycin, actinomycin D (Cosmegen), bleomycin (Blenoxane), daunorubicin, and daunorubicin (Cerubidine, Danuoxoxim); aromatase inhibitors include, but are not limited to: anastrozole (Arimidex) and letrozole (letrozole) (Femara); bisphosphonates include, but are not limited to: zoledronic acid (Zomet); cyclooxygenase inhibitors include, but are not limited to, celecoxib (Celebrex); estrogen receptor modulators include, but are not limited to, tamoxifen (Nolvadex) and fluvisol (Faslodex); folate antagonists include, but are not limited to, methotrexate and trimetrexate (tremetrexate); inorganic arsenic trioxide includes, but is not limited to, arsenic trioxide (Trisenox); microtubule inhibitors (e.g., taxanes) include, but are not limited to, vincristine (ancepin), vinblastine (Velban), paclitaxel (Taxol, Paxene), vinorelbine (navelbine), epothilone B or D or derivatives of either, and discodermolide or derivatives thereof, nitrosoureas including, but not limited to, procarbazine (Matulane), lomustine, ccnu (ceebu), carmustine (BCNU, BiCNU, gliadelwifferase), and estramustine (Emcyt); nucleoside analogs include, but are not limited to, mercaptopurine, 6-MP (Purinethol), fluorouracil, 5-FU (Adrucil), thioguanine, 6-TG (thioguanine), hydroxyurea (Hydrea), cytarabine (Cytosar-U, DepoCyt), Floxuridine (FUDR), fludarabine (Fladara), pentostatin (Nipen), cladribine (cladribine, 2-CdA), gemcitabine (Gemzar), and capecitabine (Xeloda); osteoblast inhibitors include, but are not limited to, pamidronate (Aredia); platinum-containing compounds include, but are not limited to, cisplatin (Platinol) and carboplatin (Paraplatin); retinoids include, but are not limited to, tretinoin, atra (vesanoid), alitretinoin (Panretin), and bexarotene (Targretin); topoisomerase 1 inhibitors include, but are not limited to, topotecan (Hycamtin) and irinotecan (Camptostar); topoisomerase 2 inhibitors include, but are not limited to, etoposide, VP-16(Vepesid), teniposide, VM-26(Vumon), and etoposide phosphate (etophos); tyrosine kinase inhibitors include, but are not limited to, imatinib (Gleevec); various other proteins including monoclonal antibodies, peptides and enzymes, various other molecules, such as, for example, superoxide dismutase (SOD), leptin; a flavonoid; or any combination thereof.

Non-limiting examples of anti-cancer agents and biological therapies that may be used according to some embodiments may include such treatments and molecules, such as, but not limited to: administering an immunomodulatory molecule, such as, for example, a molecule selected from the group consisting of: tumor antigens, antibodies, cytokines (such as, for example, interleukins (such as, for example, interleukin 2, interleukin 4, interleukin 12), interferons (such as, for example, interferon E1, interferon D, interferon alpha), Tumor Necrosis Factor (TNF), granulocyte macrophage colony stimulating factor (GM-CSF), macrophage colony stimulating factor (M-CSF), and granulocyte colony stimulating factor (G-CSF)), tumor suppressors, chemical factors, complement components, complement component receptors, immune system accessory molecules, adhesion molecule receptors, agents that affect cellular bioenergetics, or combinations thereof.

Osteogenic factor

In another embodiment, the active agent of the methods and compositions of the present invention is a compound that induces or stimulates the formation of bone. In another embodiment, the active agent is an osteogenic factor. In another embodiment, osteogenic factor refers to any peptide, polypeptide, protein, or any other compound or composition that induces or stimulates the formation of bone. In another embodiment, the osteogenic factor induces differentiation of the bone repair cells into osteocytes, such as osteoblasts or osteocytes. In another embodiment, the osteogenic factor is selected from the group consisting of TGF- β, BMP, and BMP. In another embodiment, the osteogenic factor is encapsulated within the matrix composition of the present invention at a concentration sufficient to induce differentiation of the bone repair cells into bone forming osteocytes.

Bone resorption inhibitor

In another embodiment, the active agent of the methods and compositions of the present invention is a compound for supporting bone restoration. In another embodiment, the active agent is a bone resorption inhibitor. In another embodiment, the active agent is a bone density maintenance agent. In another embodiment, the compound is selected from the group consisting of: osteoprotegerin (OPG), BMP-2, BMP-4, Vascular Endothelial Growth Factor (VEGF), alendronate, etidronate disodium, pamidronate, risedronate, and tiludronate. In another embodiment, the compound is Osteoprotegerin (OPG), a naturally secreted induction receptor (decoy receptor) that inhibits osteoclast maturation and activity and induces osteoclast apoptosis. In another embodiment, the active agent is a bone remodeling element (bone remodeling element). A non-limiting example of a bone remodeling component is a BMP peptide. Each possibility represents a separate embodiment of the invention.

In another embodiment, the compound is a Bone Morphogenetic Protein (BMP). In another embodiment, the compound is selected from the group consisting of BMP-2 and BMP-4, which promote osteoblast activity.

In another embodiment, the compound is Vascular Endothelial Growth Factor (VEGF).

In another embodiment, the compound is an estrogen. In another embodiment, the compound is selected from the group consisting of diphosphate derivatives. In another embodiment, the diphosphate derivative is selected from the group consisting of alendronate, etidronate disodium, pamidronate, risedronate, and tiludronate.

Each compound represents a separate embodiment of the present invention.

Antifungal agent

In another embodiment, the biologically active ingredient is an antifungal agent, for example, amphotericin B cholesteryl sulfate complex, natamycin, amphotericin, clotrimazole, nystatin, amphotericin B liposome complex, fluconazole, flucytosine, griseofulvin, itraconazole, ketoconazole, benzoic acid and salicylic acid, betamethasone and clotrimazole, butenafine, carbol, ciclopirox, clioquinol and hydrocortisone, clotrimazole, econazole, methylviolet, rupriro-diiodoquinol and hydrocortisone, ketoconazole, miconazole, naftifine, nystatin and triamcinolone, oxiconazole, sodium thiosulfate, sulconazole, terbinafine, tolnaftate, triacetin, undecylenic acid and its derivatives, butoconazole, clotrimazole, sulfonamide, terconazole, and sertaconazole.

Targeting moieties

In another embodiment, the matrix composition of the methods and compositions of the present invention further comprises a targeting moiety capable of interacting with the target molecule. Preferably, the target molecule is selected from the group consisting of a collagen molecule, a fibrin molecule and heparin. In another embodiment, the target molecule is another surface molecule that forms part of the extracellular matrix (ECM) of the target cell. The ECM is produced by the cell and assembled in situ. The most important cells involved in the assembly and maintenance of the ECM are fibroblasts. The ECM contains polysaccharide chains called GAGs (glycosaminoglycans) and various protein fibers such as collagen, elastin, fibronectin and laminin.

In another embodiment, the targeting moiety is a fibronectin peptide. Fibronectin is a high molecular weight glycoprotein that binds ECM components such as collagen, fibrin, and heparin. In another embodiment, the targeting moiety is another targeting moiety capable of interacting with a target molecule selected from the group consisting of a collagen molecule, a fibrin molecule, and heparin. Each possibility represents a separate embodiment of the invention.

In another embodiment, a "fibronectin peptide" refers to full-length fibronectin. In another embodiment, the term refers to a fragment of fibronectin. In another embodiment, the fragment comprises a collagen binding region. The collagen-binding regions of the fibronectin molecule are well known in the art and are described, for example, in Hynes, RO (1990) fibronectin. Springer-Verlag and in Yamada, KM and Clark, RAF (1996), scientific matrix in the molecular and Cellular Biology of Wound Repair (temporary matrix in molecular Biology and cell Biology, eds.) (r.a.f. Clark), pages 51-93, New York: plenum Press. Each possibility represents a separate embodiment of the invention.

In another embodiment, the targeting moiety is incorporated into the matrix composition. In another embodiment, the targeting moiety is modified to impart the ability to be incorporated into a lipid matrix. In another embodiment, the modification comprises binding to a lipid moiety. A non-limiting example of a lipid moiety is Hydrogenated Phosphatidylethanolamine (HPE). However, any lipid moiety capable of being incorporated into a lipid matrix is suitable. In another embodiment, the targeting moiety can be incorporated into the lipid matrix without modification. In another embodiment, the targeting moiety is attached to the surface of the matrix composition of the present invention. In another embodiment, the targeting moiety is attached to the surface of the matrix composition or vesicle via a hydrophobic anchor (anchor) covalently attached to the targeting moiety. In another embodiment, the targeting moiety is linked to the lipid vesicle by a hydrophobic anchor. In another embodiment, the targeting moiety is included during the preparation of the pharmaceutical carrier, allowing it to be located in a deeper layer of the carrier. Each possibility represents a separate embodiment of the invention.

In another embodiment, the target molecule is collagen. Collagen is well known in the art and is described in Khoshenodi J et al (Molecular recognition in the assembly of collagen: terminal non-gelling domains are key recognition modules in the formation of triple helix promoters) J Biol chem.281 (50): 38117-21, 2006. Each possibility represents a separate embodiment of the invention.

In another embodiment, the target molecule is fibrin. Fibrin is well known in the art and is described, for example, in Valentik LV et al (fibrin fragmentation proteins. alpha.4. beta. integrin-mediated contraction of a fibrin-fibrin protein matrix. Fibronectin fragmentation promotes the α 4. beta.1 integrin-mediated contraction of fibrin-Fibronectin provisional matrix. Exp Cell Res 309 (1): 48-55, 2005) and Mosesson MW (Fibrinogen and fibrin structure and functions. JThromb Haemost 3 (8): 1894, 904, 2005). Each possibility represents a separate embodiment of the invention.

In another embodiment, the target molecule is heparin. Heparin is well known in the art and is described, for example, in Mosesson MW (Fibrinogen and fibrin structures and functions) J Thromb Haemost 3 (8): 1894-. Each possibility represents a separate embodiment of the invention.

Additional Components

In another embodiment, the matrix composition of the methods and compositions of the present invention further comprises free fatty acids. In another embodiment, the free fatty acid is an omega-6 fatty acid. In another embodiment, the free fatty acid is an omega-9 fatty acid. In another embodiment, the free fatty acid is selected from the group consisting of omega-6 fatty acids and omega-9 fatty acids. In another embodiment, the free fatty acid moiety has 14 or more carbon atoms. In another embodiment, the free fatty acid moiety has 16 or more carbon atoms. In another embodiment, the free fatty acid moiety has 16 carbon atoms. In another embodiment, the free fatty acid moiety has 18 carbon atoms. In another embodiment, the free fatty acid moiety has 16 to 22 carbon atoms. In another embodiment, the free fatty acid moiety has 16 to 20 carbon atoms. In another embodiment, the free fatty acid moiety has 16 to 18 carbon atoms. In another embodiment, the free fatty acid moiety has from 18 to 22 carbon atoms. In another embodiment, the free fatty acid moiety has 18 to 20 carbon atoms. In another embodiment, the free fatty acid is linoleic acid. In another embodiment, the free fatty acid is linolenic acid. In another embodiment, the free fatty acid is oleic acid. In another embodiment, the free fatty acid is selected from the group consisting of linoleic acid, linolenic acid, and oleic acid. In another embodiment, the free fatty acid is another suitable free fatty acid known in the art. In another embodiment, the free fatty acid adds flexibility to the base composition. In another embodiment, the free fatty acid slows the rate of release in vivo. In another embodiment, the free fatty acid improves the consistency of controlled release in vivo. In some embodiments, the fatty acid is unsaturated. In another embodiment, the free fatty acids are saturated. In another embodiment, the incorporation of saturated fatty acids having at least 14 carbon atoms increases the gel-fluid transition temperature of the resulting matrix composition.

In another embodiment, the free fatty acid is deuterated. In another embodiment, deuteration of the acyl chain of the lipid lowers the gel-fluid transition temperature.

In another embodiment, the free fatty acid is incorporated into the matrix composition. Each type of fatty acid represents a separate embodiment of the present invention.

In another embodiment, the matrix composition of the methods and compositions of the present invention further comprises tocopherol. In another embodiment, the tocopherol of the methods and compositions of the present invention is E307 (alpha-tocopherol). In another embodiment, the tocopherol is beta-tocopherol. In another embodiment, the tocopherol is E308 (gamma-tocopherol). In another embodiment, the tocopherol is E309 (delta-tocopherol). In another embodiment, the antibiotic is selected from the group consisting of alpha-tocopherol, beta-tocopherol, gamma-tocopherol, and delta-tocopherol. In another embodiment, the tocopherol is incorporated into the matrix composition. Each possibility represents a separate embodiment of the invention.

In another embodiment, the matrix composition of the methods and compositions of the present invention further comprises a physiologically acceptable buffer salt well known in the art. A non-limiting example of a physiologically acceptable buffer salt is phosphate buffer. Typical examples of phosphate buffers are 40 parts NaCl, 1 part KCl, 7 parts Na₂HPO₄·2H₂O and 1 part of KH₂PO₄. In another embodiment, the buffer salt is any other physiologically acceptable buffer salt known in the art. Each possibility represents a separate embodiment of the invention.

Release Rate and general characteristics of matrix compositions

The in vivo release time of 90% of the active ingredient of the matrix composition of the invention is preferably between 1 week and 6 months. In another embodiment, the release time is between 4 days and 6 months. In another embodiment, the release time is between 1 week and 5 months. In another embodiment, the release time is between 1 week and 5 months. In another embodiment, the release time is between 1 week and 4 months. In another embodiment, the release time is between 1 week and 3 months. In another embodiment, the release time is between 1 week and 2 months. In another embodiment, the release time is between 2 weeks and 6 months. In another embodiment, the release time is between 2 weeks and 5 months. In another embodiment, the release time is between 2 weeks and 4 months. In another embodiment, the release time is between 2 weeks and 3 months. In another embodiment, the release time is between 3 weeks and 6 months. In another embodiment, the release time is between 3 weeks and 5 months. In another embodiment, the release time is between 3 weeks and 4 months. In another embodiment, the release time is between 3 weeks and 3 months. Each possibility represents a separate embodiment of the invention.

Methods for modulating the release rate of biodegradable polymer implants (in the absence of lipids) and drug-containing vesicles (in the absence of biodegradable polymers) are well known in the art. For example, in the case of polymers, increasing the lactic-to-co-glycolic acid ratio of PLGA will extend the release time. In the case of drug-containing vesicles, increasing the amount of cholesterol will prolong the release time. Each of these methods may be used to modulate the release rate of the matrix composition of the present invention.

As used herein, "biodegradable" refers to a substance that is capable of being broken down by natural biological processes at physiological pH. "physiological pH" refers to the pH of body tissue, typically between 6 and 8. "physiological pH" does not refer to the highly acidic pH of gastric juice, which is typically between 1 and 3.

To achieve lipid saturation, the weight ratio of total lipid to polymer can be determined by a number of methods, as described herein. In another embodiment, the lipid to polymer weight ratio of the composition of the invention is between 1: 1 and 9:1, inclusive. In another embodiment, the ratio is between 2: 1 and 9:1, inclusive. In another embodiment, the ratio is between 3: 1 and 9:1, inclusive. In another embodiment, the ratio is between 4: 1 and 9:1, inclusive. In another embodiment, the ratio is between 5:1 and 9:1, inclusive. In another embodiment, the ratio is between 6: 1 and 9:1, inclusive. In another embodiment, the ratio is between 7: 1 and 9:1, inclusive. In another embodiment, the ratio is between 8:1 and 9:1, inclusive. In another embodiment, the ratio is between 1.5:1 and 9:1, inclusive. Each possibility represents a separate embodiment of the invention.

In another embodiment for illustrative purposes, where the polymer is primarily 40KDa PLGA (poly (lactic-co-glycolic acid, 1: 1 ratio)), the molar ratio of total lipid to 40KDa PLGA is typically in the range of 20-100, inclusive. In another embodiment, the molar ratio of total lipid to 40KDa PLGA is between 20 and 200, inclusive. In another embodiment, the molar ratio is between 10 and 100, inclusive. In another embodiment, the molar ratio is between 10 and 200, inclusive. In another embodiment, the molar ratio is between 10 and 50, inclusive. In another embodiment, the molar ratio is between 20 and 50, inclusive. Each possibility represents a separate embodiment of the invention.

In another embodiment, the melting temperature (T) of the matrix composition of the invention_m) Is at least 37 ℃. In another embodiment, T_mIs at least 40 ℃. In another embodiment, T_mIs at least 42 ℃. In another embodiment, T_mIs at least 44 ℃. In another embodiment, T_mIs at least 46 ℃. In another embodiment, T_mIs at least 48 ℃. In another embodiment, T_mIs at least 50 ℃. Each possibility represents a separate embodiment of the invention.

Implants and other pharmaceutical compositions

In another embodiment, the matrix composition of the invention is in the form of an implant after evaporation of the organic solvent. The evaporation of the solvent is usually carried out at a temperature in the range of 20 to 60 ℃.

In another embodiment, the implant is homogeneous. In another embodiment, the implant is prepared by a process that includes the step of lyophilizing the material in a mold. Each possibility represents a separate embodiment of the invention.

In another embodiment, the present invention provides an implant comprising the matrix composition of the present invention comprising an antibiotic. In another embodiment, the present invention provides an implant comprising a matrix composition of the present invention comprising an NSAID. In another embodiment, the present invention provides an implant comprising the matrix composition of the present invention comprising a bone resorption inhibitor. In another embodiment, the present invention provides an implant comprising a matrix composition of the present invention comprising an antibiotic and an NSAID. In another embodiment, the invention provides an implant comprising a matrix composition of the invention comprising an antibiotic and a bone resorption inhibitor. In another embodiment, the present invention provides an implant comprising a matrix composition of the present invention comprising a bone resorption inhibitor and an NSAID. In another embodiment, the present invention provides an implant comprising a matrix composition of the present invention comprising an antibiotic, an NSAID and a bone resorption inhibitor. Each possibility represents a separate embodiment of the invention.

In another embodiment, the process of producing an implant from the composition of the invention comprises the steps of: (a) producing a matrix composition in the form of a bulk material according to the method of the invention; (b) transferring the bulk material into a mold or solid container having a desired shape; (c) freezing the loose material; and (d) freeze-drying the bulk material.

In another embodiment, the invention provides a pharmaceutical composition comprising the matrix composition of the invention and a pharmaceutically acceptable excipient.

In another embodiment, the matrix composition of the invention is in the form of microspheres after evaporation of the organic solvent. In another embodiment, the microspheres are homogeneous. In another embodiment, the microspheres are prepared by a process comprising the step of spray drying. Each possibility represents a separate embodiment of the invention.

In another embodiment, the present invention provides microspheres prepared from the matrix composition of the present invention. In another embodiment, the invention provides a pharmaceutical composition comprising the microspheres of the invention and a pharmaceutically acceptable excipient. In another embodiment, the pharmaceutical composition is in a parenterally injectable form. In another embodiment, the pharmaceutical composition is in an undissolved form. In another embodiment, the excipient is compatible with injection. In another embodiment, the excipient is compatible with infusion solutions. Each possibility represents a separate embodiment of the invention.

In another embodiment, the microspheres of the present invention have a particle size of about 500-2000 nm. In another embodiment, the particle size is about 400 and 2500 nm. In another embodiment, the particle size is about 600 and 1900 nm. In another embodiment, the particle size is about 700-1800 nm. In another embodiment, the particle size is about 500-1800 nm. In another embodiment, the particle size is about 500-1600 nm. In another embodiment, the particle size is about 600 and 2000 nm. In another embodiment, the particle size is about 700 and 2000 nm. In another embodiment, the particles are of any other size suitable for drug administration. Each possibility represents a separate embodiment of the invention.

Method of treatment

In another embodiment, the present invention provides a method of administering an antibiotic to a subject in need thereof, the method comprising the step of administering to the subject a matrix composition of the present invention, thereby administering the antibiotic to the subject in need thereof. In another embodiment, a pharmaceutical composition comprising the matrix composition is administered. In another embodiment, an implant comprising the matrix composition is administered. In another embodiment, an injectable formulation comprising the matrix composition is injected. Each possibility represents a separate embodiment of the invention.

In another embodiment, the present invention provides a method of administering a non-steroidal anti-inflammatory drug (NSAID) to a subject in need thereof, the method comprising the step of administering to the subject a matrix composition of the present invention thereby administering the NSAID to the subject in need thereof. In another embodiment, a pharmaceutical composition comprising the matrix composition is administered. In another embodiment, an implant comprising the matrix composition is administered. In another embodiment, an injectable formulation comprising the matrix composition is injected. Each possibility represents a separate embodiment of the invention.

In another embodiment, the present invention provides a pharmaceutical composition for administering an antibiotic to a subject in need thereof, comprising the matrix composition of the present invention. In another embodiment, the pharmaceutical composition is an implant. In another embodiment, the pharmaceutical composition is an injectable composition. Each possibility represents a separate embodiment of the invention.

In another embodiment, the present invention provides a pharmaceutical composition for administering an NSAID to a subject in need thereof, comprising the matrix composition of the present invention. In another embodiment, the pharmaceutical composition is an implant. In another embodiment, the pharmaceutical composition is an injectable composition. Each possibility represents a separate embodiment of the invention.

In another embodiment, the present invention provides a pharmaceutical composition for co-administering an antibiotic and an NSAID to a subject in need thereof, comprising the matrix composition of the present invention. In another embodiment, the pharmaceutical composition is an implant. In another embodiment, the pharmaceutical composition is an injectable composition. Each possibility represents a separate embodiment of the invention.

In another embodiment, the present invention provides a method of treating periodontitis in a subject in need thereof, comprising the step of administering to the subject a matrix composition of the present invention, thereby treating periodontitis. "periodontitis" refers to inflammatory diseases that affect the tissues surrounding and supporting teeth. In another embodiment, periodontitis involves the progressive loss of alveolar bone around the teeth and can eventually lead to loosening and subsequent loss of teeth if left untreated. Periodontitis has in some cases a bacterial etiology. In another embodiment, the periodontitis is chronic periodontitis. In another embodiment, the periodontitis is any other type of periodontitis known in the art. Each possibility represents a separate embodiment of the invention.

In another embodiment, the present invention provides a method of stimulating bone augmentation in a subject in need thereof, the method comprising the step of administering to the subject a matrix composition of the present invention, thereby stimulating bone augmentation. In another embodiment, the subject has a disease or disorder selected from the group consisting of: osteosarcoma/malignant fibrous histiocytoma of bone (PDQ), osteosarcoma, chondrosarcoma, ewing's sarcoma, malignant fibrous histiocytoma, fibrosarcoma and malignant fibrous histiocytoma, giant cell tumor of bone, chordoma, lymphoma, multiple myeloma, osteoarthritis, paget's bone disease, arthritis, degenerative changes, osteoporosis, osteogenesis imperfecta, bony spur, renal osteodystrophy, hyperparathyroidism, osteomyelitis, endogenetic chondroma, osteochondrosis, osteopetrosis, bone and joint disorders associated with diabetes. In another embodiment, the matrix composition is in the form of an implant. Each possibility represents a separate embodiment of the invention.

In another embodiment, the present invention provides a method of reducing the incidence of complications from orthopedic surgery in a subject in need thereof, the method comprising the step of administering to the subject a matrix composition of the present invention, thereby reducing the incidence of complications from orthopedic surgery. In another embodiment, the orthopedic surgery is selected from the group consisting of: surgery, shoulder and elbow surgery, total joint reconstruction (arthroplasty), pediatric orthopedic surgery, foot and ankle surgery, spinal surgery, arthroscopy of the knee, meniscectomy of the knee, arthroscopy of the shoulder, shoulder decompression, carpal tunnel relief, chondroplasty of the knee, removal of a carrier implant, reconstruction of the anterior cruciate ligament of the knee, replacement of the knee, repair of fractures of the femoral neck, repair of fractures of the trochanter, debridement of skin, muscle or bone fractures, repair of knee discs, hip replacement, arthroscopy of the shoulder/distal clavicle, repair of rotator tendon groups, repair of fractures of the radius/ulna, laminectomy, repair of fractures of the ankle (of the double ankle type), arthroscopy of the shoulder and debridement, fusion of the distal fracture of the radius, surgery of the intervertebral disc of the lower back, incision of the tendon sheath of the finger (lumbar spine fracture), repair of fractures of the ankle (fibula), Femoral shaft fracture repair and trochanteric fracture repair. In another embodiment, the matrix composition is in the form of an implant. In another embodiment, the implant is administered during orthopedic surgery. Each possibility represents a separate embodiment of the invention.

In another embodiment, the present invention provides a method of enhancing the effectiveness of a surgical regeneration procedure in a subject in need thereof, said method comprising the step of administering to said subject a matrix composition of the present invention, thereby enhancing the effectiveness of the surgical regeneration procedure. In another embodiment, the surgical regenerative procedure is a periodontal disease procedure (periodontological procedure). In another embodiment, the surgical regeneration procedure comprises administering an implant (implantation procedure). In another embodiment, the implantation procedure involves a spine or sinus augmentation. In another embodiment, the matrix composition is in the form of an implant. In another embodiment, the implant is administered during surgery. Each possibility represents a separate embodiment of the invention.

In another embodiment, the present invention provides a method of treating osteomyelitis in a subject in need thereof, said method comprising the step of administering to said subject a matrix composition of the present invention, thereby treating osteomyelitis. In another embodiment, the matrix composition is in the form of an implant. In another embodiment, the implant is administered at or near the site of osteomyelitis. Each possibility represents a separate embodiment of the invention.

In another embodiment, the matrix composition of the present invention is administered to aid in orthopedic bone and soft tissue restoration. The compound is administered during or after a procedure selected from the group consisting of: arthroscopic and meniscectomy, arthroscopic shoulder and decompression, carpal tunnel release, arthroscopic knee and chondroplasty, removal of carrier implants, arthroscopic knee and anterior cruciate ligament reconstruction, knee replacement, fracture repair of the femoral neck, repair of trochanter fractures, skin/muscle/bone/laceration debridement, repair of meniscal arthroscopic knee, hip replacement, arthroscopic shoulder/distal clavicle resection, repair of rotator tendon groups, repair of radius/ulna fractures, laminectomy, repair of ankle fractures (double ankle type), arthroscopic shoulder and debridement, lumbar fusion, repair of distal radius fractures, surgery of the lower dorsal disc, incision of finger tendon sheaths (incise finger tendon sheath), repair of ankle fractures (fibula), repair of fracture of the femoral shaft, and repair of trochanter fractures.

In another embodiment, the matrix composition of the present invention is applied for purposes of homeostasis, reduction of infection, and avoidance of tissue sticking by the use of products such as sponges and films.

In another embodiment, the matrix composition of the present invention is administered in order to reduce inflammatory reactions around the suture material.

Method for preparing a matrix composition

To obtain the compositions of the present invention, any suitable method that will result in a homogeneous dispersion of the polymer and lipid in a water-resistant matrix may be used. Advantageously, according to some embodiments, the method used avoids the use of water at any stage of the preparation process.

According to some embodiments, in one aspect the polymer is separately mixed with a suitably selected volatile organic solvent, and the phospholipid and the active pharmaceutical agent are mixed together with their suitably selected solvent or solvents, and then mixed together with the polymer.

In certain embodiments, the present invention provides a method of producing a matrix composition, the method comprising the steps of:

(a) mixing into a first volatile organic solvent: (i) a biodegradable polyester and (ii) a sterol; and

(b) separately mixing into a second volatile organic solvent: (i) an active agent; (ii) (ii) phosphatidylcholine and optionally (iii) phosphatidylethanolamine; and

(c) the products resulting from steps (a) and (b) are mixed and homogenized.

In another embodiment, phosphatidylethanolamine is included in the volatile organic solvent of step (a) instead of or in addition to the phosphatidylethanolamine added to the volatile organic solvent of step (b). In another embodiment, the biodegradable polyester is selected from the group consisting of PLA, PGA, and PLGA. In another embodiment, the biodegradable polyester is any other suitable biodegradable polyester known in the art. In some embodiments, the first volatile organic solvent is a non-polar solvent. In some embodiments, the second volatile organic solvent is a water-miscible solvent. In the case where the active agent is a protein or peptide, it is important to select a solvent that will not denature the protein or impair the activity of the protein. In particular embodiments, the active agent is selected from the group consisting of: NSAIDs, antibiotics, antifungal agents, steroids, anticancer agents, osteogenic factors and bone resorption inhibitors and mixtures thereof.

In another embodiment, the mixture of step (a) comprising the volatile organic solvent is homogenized before mixing it with the solution of step (b). In another embodiment, the volatile organic solvent or mixture of volatile organic solvents used in step (a) may be the same as or different from the volatile organic solvent or mixture of volatile organic solvents used in step (b). In another embodiment, the mixture of step (b) is homogenized before it is mixed with the mixture of step (a). In another embodiment, the polymer in the mixture of step (a) is lipid saturated. In another embodiment, the matrix composition is lipid saturated. Preferably, the polymer and phosphatidylcholine are incorporated into the matrix composition. In another embodiment, an active agent is also incorporated into the matrix composition. In another embodiment, the matrix composition is in the form of a lipid-saturated matrix, the shape and boundaries of which are defined by the biodegradable polymer. Each possibility represents a separate embodiment of the invention.

In another embodiment, the phosphatidylethanolamine of the methods and compositions of the present invention has a saturated fatty acid moiety. In another embodiment, the fatty acid moiety has at least 14 carbon atoms. In another embodiment, the fatty acid moiety has 14 to 18 carbon atoms. Each possibility represents a separate embodiment of the invention.

In another embodiment, the phosphatidylcholine of the methods and compositions of the present invention has saturated fatty acid moieties. In another embodiment, the fatty acid moiety has at least 14 carbon atoms. In another embodiment, the fatty acid moiety has at least 16 carbon atoms. In another embodiment, the fatty acid moiety has 14 to 18 carbon atoms. In another embodiment, the fatty acid moiety has 16 to 18 carbon atoms. Each possibility represents a separate embodiment of the invention.

In another embodiment, the weight ratio of total lipid to polymer in the first volatile organic solvent is such that the polymer in the mixture is lipid saturated. In another embodiment for illustrative purposes, where the polymer is primarily 50KDa PLGA (poly (lactic-co-glycolic acid, 1: 1 ratio)), the molar ratio of total lipid to 50KDa PLGA is typically in the range of 10-50, inclusive. In another embodiment, the molar ratio of total lipid to 50KDa PLGA is between 10 and 100, inclusive. In another embodiment, the molar ratio is between 20 and 200, inclusive. In another embodiment, the molar ratio is between 20 and 300, inclusive. In another embodiment, the molar ratio is between 30 and 400, inclusive. Each possibility represents a separate embodiment of the invention.

Each of the components of the above and other methods of the invention are defined in the same manner as the corresponding components of the matrix composition of the invention.

In another embodiment, step (a) of the production method further comprises adding phosphatidylethanolamine to the volatile organic solvent. In another embodiment, the phosphatidylethanolamine is the same as the phosphatidylethanolamine included in step (b). In another embodiment, the phosphatidylethanolamine is a different phosphatidylethanolamine, which may be any other phosphatidylethanolamine known in the art. In another embodiment, the phosphatidylethanolamine is selected from the group consisting of the phosphatidylethanolamine of step (b) and a different phosphatidylethanolamine. Each possibility represents a separate embodiment of the invention.

In another embodiment, step (a) of the production process further comprises adding tocopherol to the volatile organic solvent.

In another embodiment, step (b) of the production method further comprises adding a physiologically acceptable buffer salt to the volatile organic solvent. A non-limiting example of a physiologically acceptable buffer salt is phosphate buffer. Typical examples of phosphate buffers are 40 parts NaCl, 1 part KCl, 7 parts Na₂HPO₄·2H₂O and 1 part of KH₂PO₄. In another embodiment, the buffer salt is any other physiologically acceptable buffer salt known in the art. Each possibility represents a separate embodiment of the invention.

In another embodiment, step (b) of the production method further comprises adding a phospholipid selected from the group consisting of: phosphatidylserine, phosphatidylglycerol, sphingomyelin, and phosphatidylinositol.

In another embodiment, step (b) of the production method further comprises adding the sphingolipid to a volatile organic solvent. In another embodiment, the sphingolipid is a ceramide. In another embodiment, the sphingolipid is sphingomyelin. In another embodiment, the sphingolipid is any other sphingolipid known in the art. Each possibility represents a separate embodiment of the invention.

In another embodiment, step (b) of the production process further comprises adding the omega-6 or omega-9 free fatty acids to a volatile organic solvent that is miscible with water. In another embodiment, the free fatty acid has 16 or more carbon atoms. Each possibility represents a separate embodiment of the invention.

In another embodiment, each step of the production process is substantially free of an aqueous solution. In another embodiment, each step is substantially free of water or any aqueous solution. As provided herein, the matrix composition of the present invention produced in an anhydrous process makes it possible to saturate lipids.

Because the polymer is lipid saturated in the mixture of step (a), after mixing, a homogeneous mixture is formed. In another embodiment, the homogeneous mixture takes the form of a homogeneous liquid. In another embodiment, the vesicles are formed after lyophilization or spray drying of the mixture. Each possibility represents a separate embodiment of the invention.

In another embodiment, the production process further comprises the step of evaporating the solvent present in the product of step (c). In another embodiment, evaporation uses atomization of the mixture. In another embodiment, the mixture is atomized into dry, heated air. Typically, atomization into heated air immediately evaporates all of the water, eliminating the need for a subsequent drying step. In another embodiment, the mixture is atomized to an anhydrous solvent. In another embodiment, the evaporation is performed by spray drying. In another embodiment, the evaporation is performed by lyophilization. In another embodiment, liquid nitrogen is used for evaporation. In another embodiment, evaporation is performed using liquid nitrogen that has been premixed with ethanol. In another embodiment, evaporation is performed using another suitable technique known in the art. Each possibility represents a separate embodiment of the invention.

In another embodiment, the method of the present invention further comprises the step of vacuum drying the composition. In another embodiment, the step of evaporating is followed by a step of vacuum drying. Each possibility represents a separate embodiment of the invention.

In another embodiment, the process of the present invention further comprises the step of evaporating the organic volatile solvent by heating the product of step (c). Heating is continued at a typical temperature between room temperature and 60 ℃ until the solvent disappears. In another embodiment, the step of evaporating the solvent is followed by a step of vacuum drying. Each possibility represents a separate embodiment of the invention.

Lipid saturation and techniques for determining same

As used herein, "lipid saturation" refers to the polymer of the matrix composition being saturated with phospholipids and any other lipids that may be present that bind any hydrophobic drugs and targeting moieties present in the matrix. As described herein, in some embodiments, the matrix compositions of the present invention comprise phospholipids other than phosphatidylcholine. In other embodiments, the matrix composition comprises lipids other than phospholipids. The matrix composition is saturated with any lipids present. "saturation" refers to a state in which the matrix contains the maximum amount of the type of lipid used that can be incorporated into the matrix. Determination of the Polymer: methods of lipid ratios to achieve lipid saturation and methods of determining the degree of lipid saturation of a matrix are described herein. Each possibility represents a separate embodiment of the invention.

In another embodiment, the matrix composition of the methods and compositions of the present invention is substantially anhydrous. In another embodiment, "substantially anhydrous" refers to a composition comprising less than 1% water by weight. In another embodiment, the term refers to a composition comprising less than 0.8% by weight of water. In another embodiment, the term refers to a composition comprising less than 0.6% by weight of water. In another embodiment, the term refers to a composition comprising less than 0.4% by weight of water. In another embodiment, the term refers to a composition comprising less than 0.2% by weight of water. In another embodiment, the term refers to the absence of an amount of water that affects the water resistance properties of the composition. In another embodiment, the term refers to a composition prepared without any aqueous solvent. In another embodiment, lipid saturation can be achieved using a substantially anhydrous process to produce a composition as described herein. Lipid saturation gives the matrix composition the ability to resist substantial degradation in vivo; thus, the matrix composition exhibits the ability to modulate extended release over a range of weeks or months. Each possibility represents a separate embodiment of the invention.

In another embodiment, the matrix composition is dry. In another embodiment, "dry" refers to the absence of a detectable amount of water or organic solvent.

In another embodiment, the water permeability (water permeability) of the matrix composition has been minimized. By "minimizing" water permeability is meant the process of producing a matrix composition in an organic solvent in the presence of an amount of lipid that has been determined to minimize the permeability of the permeation of added water, as described herein. The amount of lipid required can be determined by hydrating the vesicles with a solution containing tritiated water, as described herein.

In another embodiment, "lipid saturation" refers to the filling of an internal gap (free volume) within the lipid matrix defined by the outer boundaries of the polymer backbone. The interstices are filled to such an extent by phospholipids and other types of lipids, hydrophobic drugs and targeting moieties present in the matrix: the additional phospholipid moiety can no longer be incorporated into the matrix to an appreciable extent.

In one embodiment, the following method is used to determine the degree of lipid saturation:

after preparation, the vesicles are hydrated and separated by centrifugation or filtration. The lipids not entrapped (entrap) in the vesicles form free micelles or liposomes and are located in the supernatant. The total lipid content of the supernatant and the vesicles was quantified. In this way, it was determined that different lipids were contained at the beginning: polymer ratio of encapsulated lipid to free lipid content of various formulations. Thus, the actual, experimental, maximum lipid/polymer ratio was determined.

In another embodiment, the following method is used to determine the degree of lipid saturation:

after preparation, the vesicles are hydrated with a solution containing tritiated water, washed with a tritium-free solution, and separated by centrifugation or filtration, and the amount of water included per polymer mass is quantified. With different lipids: the process is repeated for the polymer ratio to determine the amount of lipid needed to saturate the net volume of the polymer vesicles.

By "zero order release rate" or "zero order release kinetics" is meant a constant, linear, continuous, sustained and controlled release rate of the pharmaceutically active agent from the polymer matrix, i.e., the amount of pharmaceutically active agent released is linear over time.

Experimental details section

Abbreviations used: phosphoethanolamine (PE); phosphatidylcholine ═ PC; 1, 2-dimyristoyl-sn-glycero-3-phosphoethanolamine ═ DMPE (14: 0); 1, 2-dipalmitoyl-sn-glycero-3-phosphoethanolamine ═ DPPE (16: 0); 1, 2-distearoyl-sn-glycero-3-phosphocholine ═ DSPC (18: 0); 1, 2-dipalmitoyl-sn-glycero-3-phosphocholine ═ DPPC (16: 0); 1, 2-dioleoyl-sn-glycero-3-phosphocholine ═ DOPC (18: 1); 1-palmitoyl-2- {6- [ (7-nitro-2-1, 3-benzoxadiazol-4-yl) amino ] hexanoyl } -sn-glycerol-3-phosphocholine ═ NBD-PC.

Example 1Platform technology for the preparation of pharmaceutical carrier compositions:

overview

To produce a lipid-saturated polymer matrix, two mixtures were produced.

1. The biodegradable polymer and sterol and/or phospholipid components are mixed with a volatile organic solvent, which is mixed to produce a solution or suspension of the lipid-saturated polymer matrix, as measured by its Differential Scanning Calorimetry (DSC) curve.

2. The active agent and phospholipid components are mixed with a second volatile solvent to produce a second solution or suspension.

3. Combining the two solutions or suspensions and mixing until equilibrium is reached; the organic solvent is then evaporated, yielding a lipid-saturated polymer matrix containing the drug.

Experimental protocol

1. Preparation of the first solution

The polymer (PLGA, PGA, PLA or a combination thereof) and a polar lipid such as a sterol (e.g., cholesterol) and/or alpha or gamma tocopherol and/or phosphatidylethanolamine are mixed into a volatile organic solvent (e.g., ethyl acetate with/without chloroform). The mixture was mixed. The entire process is usually carried out at room temperature. Thus obtaining a first lipopolymer mixture.

Preparation of the second solution

Mixing the following materials with a volatile organic solvent (typically N-methylpyrrolidone [ NMP ], methanol, ethyl acetate, or a combination thereof)

a. Active compound

b. Phosphorylcholine or phosphatidylcholine derivatives, for example, 1, 2-distearoyl-sn-glycero-3-phosphorylcholine (DSPC) or dioleoyl-phosphatidylcholine (DOPC), are present at 30-90 mass% of all lipids in the matrix, i.e. 30-90 mass% of phospholipids, sterols, ceramides, fatty acids, etc.

c. In some experiments, phosphatidylethanolamine, such as Dimethyldimyristoylphosphatidylethanolamine (DMPE) or Dipalmitoylphosphatidylethanolamine (DPPE), was present at 0.1-50% by mass of all lipids in the matrix.

d. In some experiments, targeting moieties, e.g., fibronectin-Hydrogenated Phosphatidylethanolamine (HPE) complexes, were included as 0.1-10 mol% of all lipids in the matrix. To form this complex, fibronectin or a fragment thereof, including the collagen binding domain, is bound to the amine head group of HPE via a thioether bond.

e. In some experiments, 0.1-15 mass% of free fatty acids, such as linoleic acid (LN) or Oleic Acid (OA), are included as 0.1-10 mass% of all lipids in the matrix.

f. In some experiments, salts, such as phosphate, were included.

The second mixture is mixed, homogenized or sonicated. In some cases, a non-polar volatile organic solvent, such as ethyl acetate, is included in the mixture, which is gently stirred for 30 minutes, prior to mixing, homogenization, or sonication. The whole process is usually carried out at room temperature, but higher temperatures up to 45 ℃ are usually used when highly saturated lipids are used.

The mixture requires no water.

III-mixing of Polymer with drug/protein mixture

The second suspension (or solution) is added to the first solution with stirring. Stirring was continued for up to 5 h. The entire process is carried out at room temperature and up to 60 ℃, depending on the particular formulation, the nature of the lipid used and the particular drug. The resulting mixture will be homogeneous.

IV-Evaporation of the solvent

In some experiments, the solution from stage III was atomized into dry, heated air.

In other experiments, the solution from stage III was atomized into ethanol covered with liquid nitrogen or liquid ammonia without ethanol, after which the nitrogen and/or ethanol (as above) was evaporated.

In other experiments, when coating a surface is performed, the suspension from stage III is mixed with the particles or devices to be coated, followed by evaporation of the volatile organic solvent. The entire coating process is carried out at a temperature of 30-60 ℃.

V-vacuum drying

The coated particles and coated equipment were vacuum dried for storage.

Example 2: preparation of doxycycline hydrochloride-bone particle filler formulation for treatment of bone infection (osteomyelitis caused by trauma or bone filling effects)

1. Preparation of the first solution

The following materials were mixed into ethyl acetate:

-50-75kDa PLGA (lactic glycolic acid copolymer, 85:15 ratio)

Cholesterol-50% -100% w/w of PLGA.

The mixture was mixed. The whole process was carried out at room temperature. Thus obtaining the fat-polymer composite matrix.

Preparation of the second solution

The following materials were mixed with volatile organic solvents (methanol and ethyl acetate)

a. Active compound, antibiotic doxycycline hydrochloride

b. Phosphatidylcholine, -1, 2-distearoyl-sn-glycero-3-phosphocholine (DSPC) is present at 300-700% w/w of PLGA.

The mixture was mixed well. The whole process was carried out at room temperature. When using phosphatidylcholine (e.g., DSPC) with long saturated fatty chains, the process is typically carried out at higher temperatures of about 40-50 ℃.

The mixture requires no water.

III-mixing the Polymer with the drug mixture

The second solution or suspension is typically added to the first suspension with stirring. Stirring was continued for 1-5 minutes. The whole process is carried out at a temperature of 20-50 ℃ depending on the lipid used.

IV-surface coating followed by evaporation

To coat the bone filler particles, the particles are added to the phase III mixture, followed by evaporation of the volatile organic solvent. The whole process is carried out at a temperature of 40-50 ℃.

The ratio between the volume and percentage of solids in the phase III mixture and the mass of the bone particles will determine the time of release of the drug after hydration of the coated particles.

V-vacuum drying

The coated bone particles were vacuum dried for storage.

Bone filler particles coated with the matrix composition of the invention (commercial xenograft bovine product)) The ability of sustained release of the drug (doxycycline hydrochloride) is shown in figures 1 and 2. The rate of sustained release of doxycycline hydrochloride encapsulated within the matrix composition of the present invention was measured and compared to the release rate of the drug from bone particles treated with a free DOX solution (with the same amount of DOX). The following are found: the drug was released constantly for about three weeks (fig. 1A and B) following zero order kinetics (fig. 2). According to such assumptions: the coated bone particles constituted only 20% of the total amount of bone particles used as bone filler and the drug diffused to the free volume between the bone particles after being released, the velocity values shown in fig. 2 were calculated.

The macrostructure of the bone particles before (fig. 3A) and after (fig. 3B and 3C) coating with the matrix composition of the invention was studied. As seen in fig. 3, the structure of the bone particles was not affected by the coating: furthermore, the macrostructure of the bone particles is maintained even after incubation in serum for 60 days.

The structure of the surface of the bone particles before (fig. 4A) and after (fig. 4B-E) coating with the matrix composition of the invention was studied by SEM. The matrix composition comprises DSPC, PLGA (85:15), cholesterol and the antibiotic doxycycline hydrochloride. The coated bone particles were incubated in 10% FBS at 37 ℃ for 2 months. As can be seen, the coating is homogeneous and opaque, and moreover it covers most of the bone surface. Over time, the coating gradually disappears layer by layer through surface erosion, during which time the drug is being released. Further in fig. 4E it can be observed that: after 60 days of incubation the coating had almost completely disappeared, leaving the bone surface as its original uncoated form. The ordered structure of the matrix composition of the present invention is shown in figure 5. The coated bone particles (PLGA (85:15), DPPC (16:0), and cholesterol 10%) were analyzed by electron microscopy (X18,000) and negative staining (data not shown). The light colored lines represent the polymer, while the lipids are represented by the dark filling between the polymer materials.

The effect of different polymer/lipid compositions on the release rate of a given drug was determined using fluorescein encapsulated in matrices composed of PLGA (75:25), DPPC as the main phospholipid and varying amounts of Lauric Acid (LA) and Phosphatidylethanolamine (PE) with saturated fatty acid moieties of at least 14 carbons. As seen in fig. 6, the expected release time of 90% of the encapsulated molecules is greatly affected by LA: PE content. When the LA: PE w/w% ratio varies between 10: 10 and 0: 0, respectively (relative to the total mass of the formulation), the release lasts between about 20 and about 110 days. The drug release profile from bone particles coated with a matrix formulation comprising PE was compared to the drug release profile from bone particles coated with a matrix formulation comprising cholesterol. Figure 7 shows that the release profile of DOX from PE or cholesterol containing coated bone particles behaves similarly.

Such facts can be used to achieve different clinical needs: different polymer/lipid compositions affect the release rate. For example, anti-inflammatory therapy with NSAIDs is typically short-term (e.g., several days), so full release of the drug can be achieved within 10 days by using rapidly degrading polymers such as PLGA 50:50 and 14:0 phospholipids such as DMPC, as seen in fig. 8. Conversely, when the antibiotic drug DOX was associated with a slowly degrading polymer such as PLGA 85:15 and 18:0 phospholipids such as DSPC, full release of the drug was achieved after more than 50 days (figure 8).

The release profile of DOX from coated and uncoated bone (xenograft bovine commercial BioOss) particles was determined. The coated particles were mixed with similar normal (uncoated) bone particles in a ratio of 1: 4. As a control, we tracked the release of a similar dose of free DOX from normal (uncoated) bone particles soaked with free drug. As seen in fig. 9, the release of DOX from the formulation after hydration (37 ℃, 5% serum) was not affected by the presence of uncoated particles. In comparison, a majority of the drug from uncoated bone particles soaked with DOX was released immediately after hydration (88% within 3 hours compared to about 15% of the drug released from coated bone particles during the same period). The formulations in this study included PLGA 75:25 and DPPC.

We have further shown that the duration of drug release from coated bone particles is linearly dependent on formulation quality. The release of DOX from bone particles (12 mg/sample) coated with different masses of DOX-containing matrix formulations was compared. The duration of release of 90% of the initial DOX amount in the formulation after hydration (37 ℃, 5% serum) was monitored (fig. 10). The linear dependence between the duration of drug release and the mass of the coated matrix formulation indicates that the drug is released by gradual degradation of the matrix and that the release rate is not affected by the overall mass of the formulation.

Example 3: preparation of 1, 3-Thiabendazole (TBZ) formulation:

stock solutions：

a.PLGA/ethyl acetate, 300mg/ml (SS1, 1 ml):(i) weighing 300mg PLGA (50: 50; Sigma) into a 4ml glass cup, (ii) adding 1ml ethyl acetate, (iii) vortexing (vortex) for 5 minutes, (iv) stirring at Room Temperature (RT) for 12-18 hours, (v) confirming that the polymer particles are all dissolved, (vi) dissolving in N₂Lower, close and pack with aluminum foil and keep it at RT, (vii) solution good for 1 month.

b. Cholesterol-Ethyl acetate, 30mg/ml (SS2, 1 ml):(i) weigh 30mg cholesterol (Sigma 99%) into a 4ml glass, (ii) add 1ml ethyl acetate, (iii) vortex at RT for 5 minutes, (iv) confirm that cholesterol is totally dissolved, otherwise continue to vortex for another 2 minutes, (v) at N₂Closed, wrapped with aluminum foil and kept at RT, (vi) solution good for 1 month.

c. Ethyl acetate (C)Methanol 1: 1(SS 2.1):(i) placing 10mg of ethyl acetate in a 20ml glass cup, (ii) placing 10ml of methanol in the same(ii) in one cup, (iii) vortex for 20 seconds, (iv) keep the solution at RT, (v) the solution remains good for 1 month.

d. Thiabendazole (TBZ)/Ethyl acetate methanol 1: 1, 10mg/ml (SS3, 1 ml): (i) weighing 10mg TBZ into a 4ml glass, (ii) adding 1ml SS2.1 stock solution, (iii) vortexing at RT for 5 minutes, (iv) confirming that TBZ is fully dissolved, otherwise vortexing is continued for another 2 minutes, the solution has some white turbidity, (v) vortexing at N for another 2 minutes₂Closed, wrapped with aluminum foil and kept at RT, (vi) solution good for 1 month.

Solution a (1.2 ml):

i. 1ml of SS2(CH-EA, 30mg of CH) was added to 0.2ml of SS1(PLGA/EA, 60mg of PLGA) and placed in a 4ml glass.

Vortex for 5 min at RT.

Confirm that the mixture is homogeneous and clear, otherwise go back to ii.

At N₂Closed, wrapped with aluminum foil and kept at RT.

v. the solution remained good for 1 month.

Solution a concentration: [ CH ] ═ 25 mg/ml; [ PLGA ] ═ 50 mg/ml.

Solution B (1 ml):

i. 225mg of phospholipid (14:0) was weighed into a 4ml glass.

Add 0.75ml SS3(TBZ/EA-MET, 7.5mg TBZ).

0.25ml of ethyl acetate was put into the cup.

Vortex for 2min at RT.

v. at N₂Closed, wrapped with aluminum foil and kept at RT.

Solution remained good for 1 month.

Concentration: [ phospholipid (14:0) ] -225 mg/ml, [ TBZ ] -7.5 mg/ml.

Solution C (1 ml):

i. 0.4ml of solution B was poured into a 4ml glass.

Put 0.6ml of solution a into the cup.

Vortex for 2min at RT.

Checking: the solution was a liquid at RT, with a light yellow color with some turbidity.

v. at N₂Closing the lower part and packaging with aluminum foil.

Concentration: [ CH ] ═ 15 mg/ml; [ PLGA ] ═ 30 mg/ml; [14:0] ═ 90 mg/ml; [ TBZ ] ═ 3 mg/ml.

Bone coating preparation:

i. 12.5 (+ -0.5) mg of bone particles (Bio-Os or EndoBon) were weighed into a 1.8ml glass cup;

washing the bone with purified water (1/2ml DDW); the water was drawn out with a micropipette and then evacuated for 12-18 hours.

A heating block (heating block) was prepared and heated to 45 ℃.

Heating solution C to 45 ℃ for 30 seconds confirmed that the solution was completely dissolved and homogeneous.

v. add 50. mu.l of solution C to the bone particles using a 10-100. mu.l micropipette.

Put 1.8ml of the unsealed cup into a heating block (45 ℃) for 30 minutes.

Remove from heat and close with a plug.

Rotary pump (1X 10) for (semi-closed) cup^-1Torr) was evacuated for 12 to 18 hours.

Gently separate the melted bone particles with a spatula.

Transferring the dried coated bone particles into a new 4ml glass cup;

xi. at N₂Closed, wrapped with aluminum foil and kept at RT.

Coated bone particles remained good for 1 month.

The release profile of TBZ from bone particles coated with TBZ-containing matrix composition after hydration (37 ℃, 5% serum) can be seen in figure 11.

Example 4:drug release from absorbable gelatin sponges containing the sustained release formulations of the present invention.

A solution containing PLGA-75:25, PC16:0, cholesterol 10% and doxycycline hydrochloride (DOX) 10% was injected into the center of an absorbable gelatin sponge foam cube (gelatamp. roeko). The total DOX content in the injectable preparation was 380. mu.g in 25. mu.l. The solvent was evaporated in a 37 ℃ incubator and then under vacuum overnight. As a control, the general purpose pre-wetted with a similar dose of DOX (380 μ g) solution was injected into a gelatin sponge cube. The release of DOX from the gelatin sponge cubes into the environment after hydration (37 ℃, 5% serum) was monitored and quantified by HPLC. As seen in figure 12, when in the control sample, the entire amount of DOX was released into the medium immediately, only about 40% of the DOX associated with the PLGA/PC/cholesterol formulation was released into the medium immediately after hydration, while the remaining drug was gradually released over more than 7 days.

Example 5:SEM elemental analysis of coated bone particles: bone particles coated with matrix formulation (PLGA 50:50, cholesterol and DPPC 16:0) and uncoated bone particles were analyzed by SEM elemental analysis. Elemental analysis of the surface of the coated and uncoated bone particles is summarized in tables 1 and 2 below:

element(s)	Wt％	At％
			CK	10.05	16.91
NK	02.27	03.28
			OK	43.97	55.55
NaK	00.67	00.59
			MgK	00.76	00.63
PK	11.59	07.57
			CaK	30.68	15.47
Substrate	Correction of	ZAF

Table 1: uncoated bone particle surface

Element(s)	Wt％	At％
			CK	42.28	55.73
NK	02.96	03.34
			OK	30.90	30.57
NaK	00.46	00.32
			MgK	00.39	00.25
PK	5.94	03.04
			CaK	17.08	06.75
Substrate	Correction of	ZAF

Table 2: coated bone particle surface

Elemental analysis showed that carbon (CK in tables 1 and 2) is the predominant element in the bone particles coated with the formulation of the present invention. Carbon is a major element both in the polymer used in the formulation (PLGA 50:50) and in the lipid used (DPPC 16: 0). In contrast, the content of the main elements calcium and phosphorus of ordinary bone particles (uncoated bone particles) is at least two times lower at the surface of the coated bone particles.

The progressive degradation of bone particles coated with the formulation of the invention after hydration was studied by SEM elemental analysis. The weight percent of carbon, calcium and phosphorus atoms on the surface of the coated bone particles was monitored by SEM. As seen in fig. 13, after hydration of the coated bone particles, the percentage of carbon atoms at the surface of the coated bone particles decreased over time, while the percentages of calcium and phosphorus increased over time. These results indicate that the surface of the bone particles is gradually exposed after the coated formulation is gradually degraded.

Example 6:the increased turbidity in the supernatant of the bone particles coated with the formulation of the invention correlates with the appearance of the vesicles in the supernatant.

Bone particles (TCP, artificial bone component-commodity) coated with the formulation of the invention containing doxycycline hyclate-DOX were hydrated in 5% serum at 37 ℃. After 1 hour, the bone particles were separated from the supernatant and the supernatant was analyzed by monitoring its absorbance at 520 nm. The bone particles were re-incubated in fresh 5% serum for an additional 23 hours at 37 ℃. After 23 hours, the bone particles were separated from the supernatant and the latter analyzed as previously described.

As a result:the supernatant collected from normal uncoated bone particles did not have significant turbidity. In contrast, significant turbidity was evident in the supernatant of the coated bone particles after 1h of hydration. Three types of bone particle coatings were tested: (i) a formulation of the invention comprising an antibiotic and (ii) a DPPC and antibiotic coating and (iii) a PLGA coating. With particles coated with DPPC (OD)₅₂₀nm > 3) or particles (OD) coated with a formulation of the invention₅₂₀nm 2.0) the PLGA coated bone particles showed a smaller increase in turbidity (OD)₅₂₀nm to 0.85) (FIG. 14A). After another 23 hours of incubation (hydration) under the same conditions, the turbidity was much lower than that measured after 1h and was evident only in the lipid-containing bone-coated formulations (i) and (ii)) (fig. 14B).

After 23 hours of second incubation, supernatants removed from normal uncoated bone particles as well as coated bone particles (as described above under (i), (ii), and (iii)) were further analyzed by electron microscopy (18,000 magnification) and negative staining. In the supernatants taken from bone particles coated with formulation (i) of the invention or with dppc (ii), vesicular structures of different sizes were confirmed (fig. 14C).

The properties of the material released from bone particles coated with a matrix formulation containing PLGA (85:15), DPPC (16:0) and an antibiotic Drug (DOX) were further analyzed by a Size Distributor (Malvern Instruments DST ver.5). The coated bone particles were hydrated with 5% serum and incubated at 37 ℃ for 24 hours. After 24 hours the supernatant was removed and analyzed. The released material was characterized by two particle populations with average sizes of 550.3nm and 4.2nm (fig. 14D). The zeta potential of the particles measured using the same instrument was found to be close to zero (0.0225mV) (fig. 14E). The size of the diameter of the released material and its neutral charge may indicate that these small vesicular particles consist primarily of DPPC found in the coated matrix formulation.

The turbidity experiments described indicate that the turbidity seen in the supernatant of hydrated coated bone particles after hydration is mainly controlled by the lipid content of the formulation. The turbidity increased more slowly after an initial high turbidity associated with the kinetic behavior of DOX release from bone particles coated with the formulation of the invention (fig. 2) and with the release of fluorescein phospholipid labelled with NBD (fig. 15), characterized by an initial burst followed by a slow zero-order release.

Example 7:small angle X-ray scattering analysis of bone particles coated with the matrix composition of the invention:

we have analyzed the structure of bone particles (TCP artificial bone component-commercial product) coated with a matrix composition comprising biopolymer PLGA 85:15, lipid DPPC 16:0 and the antibiotic drug doxycycline-DOX hydrochloride. The dried particles were loaded into a glass capillary and analyzed by small angle X-ray scattering.

As a result: the scattering characteristics of the bone particles coated with the above-described matrix formulation revealed that the matrix formulation had an ordered structure having several substructures (substructures) with various sizes ranging from 5nm to 40nm (fig. 16). The dried phospholipid powder was further analyzed for structure and found to have an ordered structure with a substructure of less than 5 nm. As a control, the structure of ordinary uncoated TCP particles was investigated and found not to be characterized by the presence of sub-structures smaller than 1 nm. Thus, the substructure observed in the scattering signature of the coated bone particles can be attributed to the coating material itself and not to the normal uncoated bone particles.

Example 8:differential Scanning Calorimetry (DSC) of a Polymer comprising a solution with or without Cholesterol (solution A)

The vacuum dried polymer (PLGA (75:25)) was analyzed by differential scanning calorimetry. The temperature of the polymer was increased at a rate of 5 ℃/min with or without cholesterol (Cstrl) at different polymer/cholesterol mass ratios (w/w). The typical calorimetric reaction of PLGA (without Cstrl) shows heat uptake (heat uptake) due to PLGA melting during heating up to 200 ℃. In contrast, the addition of cholesterol reduced the caloric uptake of the polymer in a dose-responsive manner, up to a level where almost no caloric uptake was shown. Narrow caloric intake at about 150 ℃ is typical for free cholesterol in this system (fig. 17A). The effect of cholesterol was not affected by the rate of heating (data not shown). Similar but lower effects are shown when other lipids, such as alpha tocopherol, are incorporated into the polymer. In contrast, the caloric uptake of the polymer upon heating was not affected by the presence of fatty acids such as mineral oil (carbon chain C12-C18) (fig. 17B).

Example 9:the metal implant is coated with the matrix formulation of the present invention. Dental implants made of titanium were coated with the matrix formulation (PLGA 85:15, DSPC 18:0, cholesterol 10% and DOX 10%) by dipping the metal in the final solution containing the matrix composition (see step III of example 1). The solvent was then evaporated in the incubator at 37 ℃ and subsequently continued to dry under vacuum overnight (fig. 18).

Example 10:preclinical testing of matrix compositions of the invention for bone restoration

Animal model:

A. tibial osteomyelitis in rabbits

B. Bacteria: staphylococcus aureus (staphylococcus aureus)

All preclinical trials were conducted according to the regulatory guidelines for israeli animal experiments and according to the institutional ethics committee.

Test a): the relevant bacterial load was determined for the model:

1. trauma to bone (as determined in test a) -10 animals.

2. The void (damaged bone) was filled with tricalcium phosphate (TCP) material and sealed with bone wax (bonewax).

3. The site is loaded with a determined amount of bacteria by injecting the determined amount of bacteria into the site.

4. The duration is-22 days. Clinical signs and body weight were monitored (3 times per week).

5. At the end of the incubation time: animal blood was used for basic hematology & biochemistry hematology (before test termination).

6. X-ray of the tibia before termination of the trial (day 20)

7. The experiment was terminated and the tibia was collected for bacteriological testing.

8. Extracting bacteria from bone and determining bacterial concentration (as described below)

Determination of the bacterial concentration in the bone marrow:bone marrow and intramedullary cavities were swabbed with a sterile cotton swab applicator for quality assurance of total culture analysis. The inoculated applicator stripe was smeared onto a blood plate and then placed into 5ml of sterile TSB. The plates and tubes were then incubated at 37 ℃ for 24h and growth recorded.

Determination of the bacterial concentration per gram of bone:placing the bone intoInto a sterile 50mL centrifuge tube and weighed. The bone was then crushed and the final product weighed. Standard 0.9% sterile saline was added at a 3: 1 ratio (3mL saline/g bone) and the suspension was vortexed for 2 min. Six 10-fold dilutions of each suspension were prepared with 0.9% sterile standard saline. Samples (20 μ Ι _) of each dilution, including the initial suspension, were placed on blood agar plates in triplicate and incubated at 37 ℃ for 24 h; colony forming units at the maximum dilution were counted for each tibial sample. The concentration of Staphylococcus aureus was calculated as CFU/g bone.

Test a) the relevant bacterial load was determined for the model:

group of

Wound healing

Adding bacteria

Number of animals

Treatment of

Duration of time

A

Test of

Positive for

Is (L)

3

TCP (control)

22 days

B

Test of

Positive for

Is (M)

3

TCP (control)

22 days

C

Test of

Positive for

Is (H)

3

TCP (control)

22 days

D

Control

Negative of

Whether or not

1

TCP (control)

22 days

Test B) determination of the bactericidal activity of the matrix composition of the invention:

1. trauma to bone (as described in trial a) -13 animals.

2. The void (damaged bone) was filled with TCP material and sealed with bone wax.

3. The site was loaded with a defined amount of bacteria by injecting a defined amount of bacteria into the site (the amount will be determined from the results of test a).

5. During the incubation time: the animal blood was used for basic hematology & biochemistry blood tests on days 7 and 16 (before termination of the test).

6. X-ray of tibia on day 1 (or 2) plus day-20 before termination of the trial.

8. Bacteria were extracted from bone and bacterial concentrations were determined: as described above for test a.

9. The local drug concentration is determined.

Test B) determination of the bactericidal activity of the base composition of the invention (BonyPid):

group of

Wound healing

Adding bacteria

Number of animals

Treatment of

Duration of time

A

Test of

Positive for

Is that

6

BonyPid

22 days

B

Test of

Positive for

Is that

6

TCP (control)

22 days

C

Control

Positive for

Whether or not

1

TCP (control)

22 days

Test C) toxicology of the matrix composition of the invention:

1. trauma to bone (as described in trial a) -24 animals.

4. The duration is-45 days. Signs and body weight were monitored (3 times per week). The termination time is determined from the X-ray results taken during the incubation time.

5. During the incubation time: the animal blood was used for basic hematology & biochemistry blood tests on days 0, 10, 30 and 45 (before termination of the test).

6. Animals will bleed on days 1, 3, 10, 16 and 30 for plasma concentration analysis.

7. X-ray of the tibia on days 2, 20, 30 and 43 before termination of the trial.

8. The experiment was terminated and the tibia was collected for histological testing.

9. Histological examination of the lesion sites in 50% of the animals (12 animals).

10. Bacteria were extracted from bone and the bacterial concentration was determined in 50% of animals (12 animals) as described above.

Test C) toxicology of the matrix composition of the invention (BonyPid):

group of

Wound healing

Adding bacteria

Number of animals

Treatment of

Duration of time

A

Test of

Positive for

Is that

6

BonyPid

45 days

C

Test of

Positive for

Is that

6

BonyPid

45 days

D

Control

Positive for

Whether or not

6

BonyPid

45 days

F

Control

Positive for

Whether or not

6

BonyPid

45 days

Example 12: preclinical testing in periodontitis animal models

In a three-stage study, experimental periodontitis was induced in pigs using a cotton bandage placed in a proximal position. Treatment of periodontitis by a combination of Scaling and Root Planning (SRP) with one of the following treatments:

topical application of matrix implants containing flurbiprofen and doxycycline at very high, medium or low doses (30, 15, 5 and 1 mg/application site; indicated VH, H, M and L, respectively).

Topical application of matrix implants containing no active agent, in amounts corresponding to the matrix including the high, medium and low doses described above (negative control).

-topical application of flurbiprofen and doxycycline in doses corresponding to the very high, medium and low doses described above, administered as free drug.

-systemic administration of flurbiprofen and doxycycline twice daily at doses corresponding to the high, medium and low doses described above. This (in combination with SRP) is considered the benchmark for treatment of periodontitis in this animal model.

No treatment (another negative control group)

The following parameters were determined in each group each day:

vector marker level in order to determine the in vivo stability of the vector in the tissue.

-levels of flurbiprofen, doxycycline and their known metabolites in the application site, surrounding tissues and circulation.

Toxicity test

-furthermore, the following indicators of efficacy are determined:

improvement of clinical parameters like depth of Probing (PD), Clinical Adhesion Level (CAL) and Bleeding On Probing (BOP).

Improvement of radiological parameters, such as cementum-the distance between enamel and alveolar bone crest.

Histological analysis

The number of pigs in each group, the length of the study and the groups in each phase of the study are listed in tables 1-3 below:

example 13: clinical trial of the matrix composition of the present invention for periodontitis

The following studies tested the safety and clinical, radiological and microbiological efficacy of the matrix compositions of the present invention when used as an adjunct to Scaling and Root Planning (SRP).

Scaling and root planing are the most common and most conservative forms of treatment for periodontal (gum) disease. Scaling is the removal of calculus and plaque that adheres to the surface of the tooth. The procedure is particularly targeted to the subgingival area along the root. The plaque is most likely to adhere to rough surfaces. For this reason, the root surface is flattened in a process called root planing. Root planing removes any remaining stones and flattens irregular areas of the root surface.

Study design was longitudinal, randomized, single blind and intra-subject. Male and female subjects aged 20-65 years, with moderate to severe chronic periodontitis or aggressive periodontitis were recruited. A detailed medical and dental history is obtained. Elimination standard: 1) concurrent systemic disorders, i.e., pregnancy or diabetes; 2) systemic antibiotics or NSAID drugs were used over the last 3 months; 3) smoking; 4) any known allergy to the ingredients of the matrix composition; 5) periodontal treatment was performed less than 6 months prior to baseline. The subject undergoes SRP either alone or in combination with administration of: (a) a matrix implant comprising an antibiotic + NSAID drug; (b) a matrix implant free of active ingredients; (c) free antibiotic + NSAID drug for oral administration; (d) systemic antibiotics + NSAIDs. Implants or free antibiotics are applied in the oral cavity at multiple sites.

The following clinical measurements were recorded at baseline and at 1, 3, 6 and 9 months: depth of Probe (PD), Clinical Attachment Level (CAL), Bleeding On Probe (BOP) and gingivitis, plaque and staining index.

Microbiological tests were performed, including bacterial culture and N-benzoyl-DL-arginine-naphthamide (BANA) test.

The foregoing description of the specific embodiments will so fully reveal the general nature of the invention that others can, by applying current knowledge, readily modify such specific embodiments for various applications without undue experimentation and without departing from the general concept, and, therefore, such modifications and improvements are intended to be comprehended within the meaning and range of equivalents of the disclosed embodiments. It is to be understood that the phraseology or terminology employed herein is for the purpose of description and not of limitation. The methods, materials, and steps for performing various disclosed functions may take a variety of alternative forms without departing from the invention.

Claims

1. A matrix composition comprising:

a. a biodegradable polymer associated with a first lipid having polar groups, wherein the first lipid comprises a sterol;

b. a second lipid comprising at least one phospholipid having a hydrocarbon chain of at least 14 carbons; and

c. a pharmaceutically active agent;

wherein the matrix composition is lipid-saturated, substantially anhydrous and has an ordered multi-layered structure in which the biodegradable polymer and lipid are ordered in the form of layers, and wherein the matrix composition is suitable for providing sustained release of the pharmaceutically active agent, wherein at least 50% of the pharmaceutically active agent is released from the matrix composition with zero order kinetics.

2. The matrix composition of claim 1, wherein the phospholipid is a phosphatidylcholine having fatty acid moieties comprising at least 14 carbons.

3. The matrix composition of claim 1, wherein the biodegradable polymer is a biodegradable polyester selected from the group consisting of: polylactic acid (PLA), polyglycolic acid (PGA), poly (lactic-co-glycolic acid) (PLGA), and combinations thereof.

4. The matrix composition of claim 1, wherein the pharmaceutically active agent is selected from the group consisting of: antibiotics, antifungal agents, non-steroidal anti-inflammatory drugs, steroids, anti-cancer agents, osteogenic factors, and bone resorption inhibitors.

5. The matrix composition of claim 4, wherein the pharmaceutically active agent is selected from an antibiotic or an antifungal agent.

6. The matrix composition of claim 4, wherein the pharmaceutically active agent is an anti-cancer agent.

7. The matrix composition according to claim 4, wherein the pharmaceutically active agent is a non-steroidal anti-inflammatory drug (NSAID).

8. The matrix composition of claim 4, wherein the pharmaceutically active agent is a steroid.

9. The matrix composition of claim 4, wherein the pharmaceutically active agent is selected from an osteogenic factor or a bone resorption inhibitor.

10. The matrix composition of claim 1, wherein the weight ratio of total lipid to the biodegradable polymer is between 1.5:1 and 9:1, inclusive.

11. The matrix composition of claim 1, comprising a plurality of pharmaceutically active agents selected from the group consisting of: antibiotics, antifungal agents, non-steroidal anti-inflammatory drugs (NSAIDs), steroids, anticancer agents, osteogenic factors, and bone resorption inhibitors.

12. The matrix composition of claim 1, wherein the matrix composition is homogeneous.

13. The matrix composition of claim 1, comprising a sphingolipid.

14. The matrix composition of claim 1, comprising tocopherol.

15. The matrix composition of claim 1, further comprising an additional phospholipid selected from the group consisting of phosphatidylserine, phosphatidylglycerol, phosphatidylinositol, or a combination thereof.

16. The matrix composition of claim 1, further comprising a free fatty acid having 14 or more carbon atoms.

17. The matrix composition of claim 1, further comprising a targeting moiety capable of interacting with a target molecule selected from the group consisting of a collagen molecule, a fibrin molecule, and heparin.

18. The matrix composition of claim 17, wherein the targeting moiety is a fibronectin peptide.

19. The matrix composition of claim 1, further comprising a pegylated lipid.

20. The matrix composition of claim 1, wherein the sterol is cholesterol.

21. The matrix composition of claim 20, wherein the cholesterol is present in an amount of 5-50 mole percent of the total lipid content of the matrix composition.

22. The matrix composition according to claim 1, for sustained release of the pharmaceutically active agent, wherein at least 60% of the pharmaceutically active agent is released from the matrix composition at zero order kinetics.

23. The matrix composition according to claim 22, for sustained release of the pharmaceutically active agent, wherein at least 65% of the pharmaceutically active agent is released from the matrix composition at zero order kinetics.

24. An implant comprising the matrix composition of claim 1.

25. A pharmaceutical composition for sustained release of an active agent comprising the matrix composition of claim 1.

26. The pharmaceutical composition of claim 25, wherein the active agent is selected from the group consisting of: antibiotics, antifungal agents, non-steroidal anti-inflammatory drugs, steroids and anticancer agents, osteogenic factors and bone resorption inhibitors.

27. Use of the matrix composition of claim 5 in the preparation of a medicament for administering an antibiotic to a subject in need thereof.

28. Use of the matrix composition according to claim 5 for the preparation of a medicament for the treatment of periodontitis.

29. Use of a matrix composition according to claim 7 in the manufacture of a medicament for administering a non-steroidal anti-inflammatory drug (NSAID) to a subject in need thereof.

30. Use of the matrix composition according to claim 7 for the preparation of a medicament for the treatment of periodontitis.

31. Use of the matrix composition of claim 9 in the preparation of a medicament for administering a bone resorption inhibitor to a subject in need thereof.

32. Use of the matrix composition of claim 9 in the preparation of a medicament for stimulating bone augmentation in a subject in need thereof.

33. A medical device, comprising: a substrate and a biocompatible coating deposited on at least a portion of the substrate, wherein the biocompatible coating comprises the matrix composition of claim 1.

34. The medical device of claim 33, wherein the biocompatible coating comprises multiple layers.

35. The medical device of claim 33, wherein the substrate comprises at least one material selected from the group consisting of: hydroxyapatite, stainless steel, cobalt-chromium, titanium alloys, tantalum, ceramics and gelatin.

36. The medical device of claim 33, wherein the substrate is selected from the group consisting of: orthopedic nails, orthopedic screws, orthopedic wires, orthopedic needles, metallic or polymeric implants, bone filler particles, collagen and non-collagen films, suture materials, orthopedic cements, and sponges.

37. The medical device of claim 36, wherein the orthopedic staples are orthopedic staples.

38. The medical device of claim 36, wherein the bone filler particles are selected from the group consisting of allogeneic, xenogeneic, and artificial bone particles.

39. Use of the substrate composition according to any one of claims 1-23 for the coating of a substrate selected from the group consisting of: orthopedic nails, orthopedic screws, orthopedic wires, orthopedic needles, metallic or polymeric implants, bone filler particles, collagen and non-collagen films, suture materials, orthopedic cements, and sponges.

40. The use according to claim 39, wherein the orthopedic staples are orthopedic staples.

41. A method of producing a matrix composition, the method comprising the steps of:

a. mixing into a first volatile organic solvent: (i) a biodegradable polymer and (ii) a first lipid having a polar group, the first lipid comprising at least one sterol;

b. mixing into a second volatile organic solvent: (i) at least one pharmaceutically active agent; (ii) a second lipid selected from phospholipids having a hydrocarbon chain of at least 14 carbons; and

c. mixing the products produced in steps (a) and (b) to produce a homogeneous mixture; and is

d. Evaporating the first volatile organic solvent and the second volatile organic solvent;

wherein each step of the process is substantially free of an aqueous solution;

thereby producing a homogeneous matrix composition, wherein the matrix composition has an ordered multi-layered structure.

42. The method of claim 41, wherein the phospholipid is a phosphatidylcholine having fatty acid moieties that contain at least 14 carbons.

43. The method of claim 41, wherein the first lipid further comprises phosphatidylethanolamine comprising fatty acid moieties having at least 14 carbons.

44. The method of claim 41, wherein the biodegradable polymer is a polyester selected from the group consisting of: polylactic acid (PLA), polyglycolic acid (PGA), poly (lactic-co-glycolic acid) (PLGA), and combinations thereof.