Method for delivering a peptide to a subject at a modulated rate via microcapsules of lactic-co-glycolic copolymer containing said peptide

Abstract

Micro-capsules for the slow release of drugs, consisting of a lactic-co-glycolic copolymer to which a plasticizer has been incorporated and which contain a drug of pharmaceutical interested within them.

Claims

1. A method for delivering a peptide analog of luteinizing hormone releasing, hormone (LHRH) selected from the group consisting of triptorelin, leuprolide, goserelin, buserelin and cetrorelix at a modulated rate to a subject in need thereof, said method comprising: administering to the subject an amount of microcapsules of a polymer selected from a polylactic acid polymer or a lactic-co-glycolic copolymer containing the peptide analog to provide a therapeutic effect, wherein the polymer incorporates triethyl citrate as an additive, wherein the triethyl citrate is added in a range of between 2.7% and 4.4% by total weight of the microcapsule components, and wherein the triethyl citrate produces a modulation of the release of the peptide analog from the microcapsules and a reduction of the initial release rate of the peptide analog from the microcapsules as compared to an identical microcapsule in the absence of triethyl citrate.

2. The method according to claim 1, wherein the polymer is a lactic-co-glycolic copolymer comprising lactate and glycolate units which are present in a ratio of between 99:1 and 10:90, both inclusive.

3. A method for delivering a peptide analog of luteinizing hormone releasing hormone (LHRH) selected from the group consisting of triptorelin, leuprolide, goserelin, buserelin and cetrorelix at a modulated rate to a subject to whom a plurality of microcapsules containing said peptide analog are administered, said method comprising: (a) forming a plurality of microcapsules containing the peptide analog by (i) dissolving a polymer selected from a polylactic acid polymer or a lactic co-glycolic copolymer and triethyl citrate in a solvent in which said polymer and said triethyl citrate are soluble to form a solution; (ii) adding the peptide analog to the solution to form a peptide suspension in said solution; (iii) adding an alkyl derivative to the solution to produce deposition of the polymer and the triethyl citrate on the peptide analog; (iv) adding the suspension obtained in step (ii) to a solution in which said polymer and triethyl citrate are not soluble to harden and precipitate microcapsules thus formed; and (v) isolating the microcapsules thus formed, wherein the polymer incorporates triethyl citrate as an additive, wherein the triethyl citrate is added in a range of between 2.7% and 4.4% by total weight of the microcapsule components, and (b) administering a sufficient amount of said microcapsules containing the peptide analog to provide a therapeutic effect to said subject, wherein the triethyl citrate produces a modulation of the release of the peptide analog from the microcapsules and a reduction of the initial release rate of the peptide analog from the microcapsules as compared to an identical microcapsule in the absence of triethyl citrate.

4. The method according to claim 3, wherein a ratio between lactate and glycolate units in the lactic co-glycolic copolymer is between 99:1 and 10:90, both inclusive.

5. The method according to claim 3, wherein the alkyl derivative is silicon oil.

6. The method according to claim 1, wherein the lactic-co-glycolic copolymer comprises lactate and glycolate units present in a ratio of 1:1.

7. The method according to claim 1, wherein the molecular weight of the lactic-co-glycolic polymer is 50,000.

8. The method according to claim 1, wherein the peptide analog of LHRH is leuprolide.

9. The method according to claim 1, wherein the peptide analog of LHRH is leuprolide acetate.

10. A microcapsule comprising: i. a peptide analog selected from leuprolide, octreotide or triptorelin; ii. a polymer selected from a polylactic acid polymer or a lactic-co-glycolic copolymer containing the peptide analog; and iii. triethyl citrate; wherein the polymer incorporates triethyl citrate as an additive, wherein the triethyl citrate is added in a range of between 2.7% and 4.4% by total weight of the microcapsule components and wherein the triethyl citrate produces a modulation of the release of the peptide analog from the microcapsules and a reduction of the initial release rate of the peptide analog from the microcapsules as compared to an identical microcapsule in the absence of triethyl citrate.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

(1) FIG. 1 shows percent leuprolide drug released by the microcapsules over time (days).

(2) The preparation of the micro-capsules can be carried out following any of the methods described in the literature such as, for example, those described in the U.S. Pat. No. 3,773,919. By way of description and without limitation thereto, the different procedures for producing micro-capsules of the invention would be grouped into the following sections:

(3) a) Method of Coacervation:

(4) A solution of polymer is prepared along with tri-ethyl citrate in a suitable solvent. The drug to be encapsulated is suspended in the polymer and plasticiser solution and a non-solvent of the polymer is added to force deposition of the polymer on the drug crystals. Examples of these procedures without using plasticiser can also be found in documents such as ES 2009346 or EP 052510.

(5) b) Double Emulsion Methods:

(6) The drug to be encapsulated is dissolved in water or in a solution of some other co-adjuvant and is emulsified in a solution of the polymer and the plasticiser in a suitable solvent such as dichloromethane, for example. The resulting emulsion is in turn emulsified in water or in an aqueous solution of an emulsifier such as polyvinylic alcohol. Once this second emulsion has been carried out, the solvent in which the polymer was dissolved is eliminated through evaporation or extraction. The resulting micro-capsules are obtained directly by filtration. Examples of these procedures that do not use the plasticiser can also be found in documents such as U.S. Pat. No. 4,652,441.

(7) c) Simple Emulsion Method:

(8) The drug to be encapsulated, the polymer and the plasticiser are dissolved together in a suitable solvent. This solution is emulsified in water or in a solution of an emulsifier such as polyvinyl acid, and the organic solvent is eliminated by evaporation or extraction. The resulting micro-capsules are recovered by filtration. Examples of these procedures that do not use the plasticiser can also be found in documents such as U.S. Pat. No. 5,445,832.

(9) d) Methods of Solvent Evaporation:

(10) The drug to be encapsulated, the polymer and the plasticiser are dissolved together in a suitable solvent. This solution is evaporated to dryness and the resulting residue reduced down to a suitable size. Examples of this procedure, although not using the plasticiser, can be also be found in documents such as GB 2,209,937.

(11) In the present invention, in all cases, the citric acid ester is deposited along with the polymer, plastifying it and advantageously modifying the hydrophobicity, flexibility and coating capacity characteristics of the polymer and the release profile of the micro-capsules obtained.

(12) This is reducing the initial release of the encapsulated drug and making this release almost linear in time.

(13) The present invention is now described by means of following, non-limiting examples:

EXAMPLE 1: PRODUCTION OF MICRO-CAPSULES, CONTAINING LEUPROLIDE ACETATE, WHICH PRESENTS A DRUG RELEASE PROFILE SUITABLE FOR ONE MONTH

(14) 3 g of tri-ethyl citrate and 1.45 g of lactic-co-glycolic polymer (mw=50000 with monomer ratio of 1/1) are dissolved in 50 ml of dichloromethane. When the polymer is fully dissolved 67 mg of leuprolide acetate are added and then suspended by sonication.

(15) 63 g of silicone of 350 cts is added slowly with intensive stirring. And when all the silicone has been added the content of the reactor is poured onto 2.5 l of n-heptane and stirred for 1 hour.

(16) The micro-capsules are recovered by filtration and dried under vacuum for 48 hours.

EXAMPLE 2: PRODUCTION OF MICRO-CAPSULES WITH ONE-MONTH RELEASE CONTAINING OCTREOTIDE ACETATE

(17) 2 g of tri-ethyl citrate and 1.45 g of lactic-co-glycolic polymer (mw=50000 with monomer ratio of 1/1) are dissolved in 50 ml of dichloromethane. When the polymer is fully dissolved 67 mg of octreotide acetate are added and then suspended by sonication.

(18) 70 g of silicone of 350 cts is added slowly with intensive stirring. And when all the silicone has been added the content of the reactor is poured onto 2.5 l of n-heptane and stirred for 1 hour.

(19) The micro-capsules are recovered by filtration and dried under vacuum for 48 hours.

EXAMPLE 3: PRODUCTION OF MICRO-CAPSULES WITH A THREE-MONTH RELEASE PROFILE CONTAINING TRIPTORELINE ACETATE

(20) 2 g of tri-ethyl citrate and 1.45 g of lactic-co-glycolic polymer (mw=50000 with monomer ratio of 1/1) are dissolved in 50 ml of dichloromethane. When the polymer is fully dissolved 45 mg of triptoreline acetate are added and then suspended by sonication.

(21) 70 g of silicone of 350 cts is added slowly with intensive stirring. And when all the silicone has been added the content of the reactor is poured onto 2.5 l of heptane and stirred for 1 hour.

(22) The micro-capsules are recovered by filtration and dried under vacuum for 48 hours.

EXAMPLE 4: IN VITRO DETERMINATION OF THE DRUG RELEASE BY THE MICRO-CAPSULES OBTAINED

(23) Material Needed:

(24) 12 plastic 10-ml tubes with lid.

(25) 1 rack for tubes.

(26) Procedure:

(27) Approximately 10 mg of micro-capsules containing leuprolide obtained according to example 1 are weighed into 12 10-ml tubes.

(28) To each tube 2 ml of phosphate buffer 1/30 M and pH=7.0 are added.

(29) Each tube is gently shaken to suspend the micro-capsules in the buffer, the tubes are sealed and placed in an oven at 37° C.

(30) Taking samples for the control of the hydrolysis is carried out in accordance with the following table:

(31) TABLE-US-00001 TABLE 1 Taking samples for analysis of leuprolide released. Tube no. Type of analysis Time  1 h 1, 2 Supernatant  3 h 3 Supernatant  6 h 4 Supernatant  1 d 5 and 6  Pellet  2 d 7 Pellet  4 d 8 Pellet Point  8 d 10  Pellet 11 d 1 and 11 Pellet 14 d 2 Pellet 18 d 3 and 12 Pellet 23 d 9 Pellet 29 d 4 and 5  Pellet

(32) The analysis of leuprolide released is carried out by HPLC in the following conditions:

(33) COLUMN: Kromasil C-8; 25×0.45 cm

(34) ELUENT: Acetonitrile/water 30/70+0.05% trifluoracetic acid

(35) FLOW RATE: 1 ml/min

(36) DETECTION: UV 280 nm.

(37) The samples are taken at the times indicated in table 1 and the analysis carried out by quantifying the peptide released in the supernatant (supernatant analysis) or the residual peptide inside the micro-capsule (pellet analysis) depending on the hydrolysis time, as indicated in table 1.

(38) The result of this analysis is indicated in FIG. 1. In this FIGURE, the results obtained are compared with a control assay performed with leuprolide microcapsules in which triethyl citrate has not been incorporated, in accordance with the method of Example 1.