A OLIGOMERIC LACTIDE MACROMER BASED COPOLYMER AND A PROCESS FOR THE PREPARATION THEREOF

Field of the invention

The present invention relates to a copolymer composition comprising a lactide macromer and a basic monomer and method for the preparation of these copolymers thereof. In particular a composition comprising an acrylate or methacrylate ester of oligomeric lactide is provided. These copolymers exhibit unusual dissolution behavior. They are soluble over wide range of pH from 1.8 to 7.4. This dissolution property can be exploited in designing pharmaceutical dosage forms.

Background of the invention Biodegradable polyesters derived from aliphatic hydroxy carboxylic acids have been developed for medical applications such as surgical sutures, drug delivery devices, tissue supports, and implants for internal bone fixation (S.W. Shalaby and A. Johnson In: S.W. Shalaby, Editor, Biomedical polymers: Designed to degrade systems, Carl Hanser Verlag, Munich (1994), pp. 1-34. , R.L. Dunn In: J. O. Hollinger, Editor, Biomedical applications of synthetic biodegradable polymers, CRC Press, Boca Raton (1995), pp. 17-31. V. Maquet and R. Jerome Mater. Sci. Forum 250 (1997), pp. 15-42).

Most of these materials are made from high-molecular-weight linear polyesters like polylactides, polyglycolides and their copolymers (D. E. Perrin and J. P. English In: A.J. Domb, J. Kost and D.M. Wiseman, Editors, Handbook of biodegradable polymers, Harwood Academic Publishers, Amsterdam (1997), pp. 3-27.) Less attention has been paid to oligomeric esters, because these oligomers normally do not have the mechanical and thermal properties required for sutures or implants. Recent work on synthesis of liquid or low melt oligolactides offers interesting approach to a new class of biodegradable materials useful for injectable drug delivery systems, implant coatings or soft tissue augmentations. In addition, biodegradable polymer networks and composites can be prepared from these oligoesters terminated with unsaturated functional groups (D.K. Han and J.A. Hubbell Macromolecules 30

(1997), pp. 6077-6083, G. Coullerez, C. Lowe, P. Pechy, H. H. Kausch and J. Hilborn J. Mater. Sci: Mater. Med. 11 (2000), pp. 505-510.)

Novel linear and star-shaped oligolactide macromers were prepared and used for the fabrication of highly porous polymer network scaffolds of controlled shape. In vitro studies on the cultivation of osteoblasts on these materials demonstrated that the polymer networks possess excellent biocompatibility and they are well suited as scaffolds for bone tissue engineering. (Matthias Schnabelrauch, Sebastian Vogta, Yves Larcherb and lngo Wilkeb, Biomolecular Engineering, 19 (2-6), (2002), pp.295-298) Controlled ring-opening polymerization of L-Lactide was initiated using cyclic tin alkoxides which resulted in series of lactide macromonomers. Double bond of the initiator was successfully incorporated into the synthesized macromonomers which is well-suited for postpolymerization into a brushlike polymer. (Ryner, M.; Finne, A.; Albertsson, A. C; Kricheldorf, H. R. Macromolecules 34, (2001) pp.7281-7287) This unsaturated macromonomer provided a variety of opportunities for further modifications. The incorporated C=C double bond was oxidized into epoxides. (Finne, Anna; Albertsson, Ann- Christine. Journal of Polymer Science, Part A: Polymer Chemistry 42(3), (2004), pp.444-452) Poly (D, L) lactide diacrylate macromer was used to develop a new family of biodegradable hydrogels with photo-crosslinked dextran derivative of allyl isocyanate The changes in thermal and mechanical properties of these hydrogels as function of dextran and lactide macromer composition were investigated. (Zhang, YeIi; Chu, Chin-Chang. Journal of Materials Science: Materials in Medicine 13(8), (2002), pp.773-781).

A series of temperature and pH-sensitive hydrogels based on poly (2- ethyl-2-oxazoline) and three-arm poly (D, L-lactide) macromer were synthesized via photo-copolymerization. Lactide macromer was synthesized by first reacting lactide with Glycerol and then reacting 3 arm poly-lactide with methacryloyl chloride and triethylamine. This study effectively proved that unique combination of water swellability and biodegradability properties provides hydrogels for a much wider range of applications in biomedical fields.

(Wang, Chau-Hui; Hsiue, Ging-Ho. Journal of Polymer Science, Part A: Polymer Chemistry 40(8), (2002), pp.1112-1121)

Difunctional oligolactone macromers were synthesized by ring-opening oligomerization of various lactones (L-lactide, glycolide, p-dioxanone) in the presence of suitable diols (propane-1 , 2-diol, dianhydro-D-glucitol) and subsequent end capping of these oligolactones with methacrylate moieties. Highly porous scaffolds were fabricated from these macromers. The oligolactide based polymer networks possess excellent biocompatibility and are promising candidates as scaffolds in bone tissue engineering. ( Haris, Parvez I.; Vogt, S.; Berger, S.; Wilke, I.; Larcher, Y.; Weisser, J.; Schnabelrauch, M. Bio-Medical Materials and Engineering 15(1 , 2), (2005), pp.73-85.)

Weak polyacids or polybases, which undergo an ionization / deionization transition from pH 4~8, are utilized as pH-responsive polymers. Poly (N, N'- dimethyl amino ethyl methacrylate) (PDMAEMA) and poly (N, λ/'-diethyl amino ethyl methacrylate) (PDEAEMA) are examples of pH responsive polybases. (Eun Seok Gil, Samuel M. Hudson. Prog. Polym. Sci. 29 (2004) pp.1173- 1222). They have amine groups in their side chains. The amine groups gain protons under acidic condition and release them under basic condition. PDMAEMA was also reported to exhibit temperature sensitivity similar to PNIPAAm (M. Okubo, H. Ahmad and T. Suzuki, Colloid Polym Sci 276 (1998), pp. 470-475) The origin of the thermo-responsive behavior lies in the balance of hydrophobicity and hydrophilicity of poly (DMAEMA), while the pH-sensitive behavior is due to the existence of tertiary amino-group, which gets protonated with decreasing pH of the aqueous medium. These properties of poly (DMAEMA) were used to develop hydrogels having good mechanical properties through copolymerization of DMAEMA with other methacrylate derivatives, among which the copolymers and hydrogels prepared from DMAEMA and Butyl methacrylate (BMA) were studied in detail. By using a unique method and photoredox system to initiate the copolymerization of DMAEMA with BMA, a stable asymmetric bilayer sheet which shows reversible thermal and pH-responsive behavior was developed as intelligent soft material.

(Li, Fu-Mian; Chen, Shuang-Ji; Du, Fu-Sheng; Wu, Zhi-Qiang; Li, Zi-Chen. ACS Symposium Series 726 (1999), pp.266-276)

A pH-sensitive interpolymer polyelectrolyte complex was synthesized by gamma radiation induced copolymerization of acrylic acid and N, N'-Dimethyl amino ethyl methacrylate (DMAEMA). pH dependent swelling showed different phase transitions depending on the copolymer composition and also showed the interpolymer polyelectrolyte complex formation at pH values ranging from pH 3 to pH 4. The ability of the copolymer to be used as drug carrier for colon specific drug delivery system was demonstrated using Ketoprofen as a model drug. (El-Hag AIi Said, Amr. Biomaterials 26(15), (2005), pp.2733-2739.)

A new pH/temperature responsive polymer system with transitions resulting both from polymer-water and polymer-polymer interactions has been demonstrated using the copolymer composed of N,N'-dimethyl amino ethyl methacrylate (DMAEMA) and ethylacrylamide (EAAm) and the mixture of poly DMAEMA and poly EAAm. Based on the pH / temperature responsiveness of the copolymer and polymer mixture, glucose controlled insulin delivery system and microspheres for temperature sensitive solute release were designed and characterized^ Yuk, Soon Hong; Seo, Jung Ki; Lee, Jin Ho; Cho, Sun Hang. ACS Symposium Series 752 (2000), pp.232-242.) The insulin release from the same copolymer matrix was demonstrated.

(Yuk, Soon Hong; Cho, Sun Hang; Lee, Sang Hoon. Polymer Preprints 39(2), (1998), pp.204-205.). Poly (N, N-dimethyl amino ethyl) methacrylate (DMAEMA) and polyethyl acrylamide (EAAm) system was also used to design the microspheres for pH / temperature sensitive drug release. Hydrocortisone was used as a model drug. This gave the control of hydrocortisone release in an on-off manner without considerable lag time (Kim, E. J.; Cho, S. H.; Yuk, S. H. Biomaterials 22(18), (2001), pp.2495-2499).

In the same way copolymers of DMAEMA and ethylacrylamide (EAAm) [or acrylamide (AAm)] were prepared and characterized as polymeric drug delivery systems modulated for pulsatile and time release. When the temperature of poly DMAEMA aqueous solution was increased above 50 ⁰ C, the polymer precipitated from the solution. The incorporation of EAAm in the

O

copolymer caused lower critical solution temperature (LCST) to shift to a lower temperature. This was because of the formation of hydrogen bonds, which protect (N, N-dimethyl amino) ethyl groups from exposure to water and led to a hydrophobic contribution to the LCST. Glucose controlled insulin release and thermosensitive permeation of hydrocortisone was accomplished by manipulating pH / temperature responsiveness of polymers. (Yuk, Soon Hong; Cho, Sun Hang; Lee, Sang Hoon; Seo, Jung Ki; Lee, Jin Ho. Editor(s): Ottenbrite, Raphael M.; Kim, Sung Wan. Polymeric Drugs & Drug Delivery Systems (2001), pp.39-55.) The effect of pendent side-chain length and crosslinking agent concentration in methyl methacrylates / dimethyl amino ethyl methacrylate as polybasic / hydrophobic pH-sensitive hydrogel was studied. Increasing both side-chain length and crosslinking agent concentration decreased the sharpness of response to pH and water-uptake capacity of the polymer. (Falamarzian, M.; Varshosaz, J. Drug Development and Industrial Pharmacy 24(7), (1998), pp.667-669).

Amphiphilic comb-copolymers made by copolymerization of N, N- dimethyl amino ethyl methacrylate (DMAEMA) and poly (Butyl acrylate) macromonomers are excellent dispersants for many organic pigments in different coating systems. (Muehlebach, Andreas. Polymeric Materials Science and Engineering 90, (2004), pp.180).The glucose-responsive insulin controlled release system based on the hydrogel poly (2-hydroxyethyl methacrylate-co- N, N-dimethyl amino ethyl methacrylate), with entrapped glucose oxidase, catalase and insulin was studied. When exposed to physiological fluids, glucose diffuses into the hydrogel; glucose oxidase catalyzes the glucose conversion to gluconic acid, causing swelling of the pH-sensitive hydrogel and subsequently increased insulin release. The effects of polymer morphology and oxygen availability on hydrogel swelling and on insulin release kinetics were tested. In vivo experiments on rats demonstrated that at least some of the entrapped insulin retained its active form and was effective in reducing blood glucose levels. Thus the pH-sensitive hydrogel poly (HEMA-co-DMAEMA) could be manipulated to produce glucose-responsive insulin release system

that was effective in reducing blood glucose levels. (Traitel, Tamar; Cohen, Yachin; Kost, Joseph. Biomaterials 21(16), (2000), pp.1679-1687.)

Copolymerization of acrylate monomers with basic monomers like Dimethyl amino ethyl methacrylate (DMAEMA) with the aim of designing controlled drug delivery of therapeutic or complex protein molecules like insulin or site specific drug delivery using hydrogels has been reported. Such polymers containing basic functional groups such as amino groups are known to dissolve at pH prevalent in the stomach. Hence these copolymers of acrylates with DMAEMA result in pH sensitive polymers and at times exhibit temperature sensitive behavior. The copolymers have high molecular weights and they are used in variety of applications like drug delivery systems and also as dispersants in pigment industry. DMAEMA along with other neutral methacrylates (Composition of Eudragit E) is also used as reverse enteric coating. This polymer shows swelling at pH 5. Percentage of DMAEMA in Eudragit E is high (35% w/w), which possibly results in negative drug interaction with some of the drugs. (MJ. Alonso, M. L Lorenzo-Lamosa, M.Cuna, J.LVila-Jato and D.Torres, Journal of Microencapsulation, 1997, Volume14, No.5, 607-616). Low molecular weight DMAEMA polymers having low basic monomer content and their utility as excipient in pharmaceutical drug delivery has not been investigated in the past.

This invention describes composition of a low molecular weight macromer based copolymer synthesized using oligomeric lactide macromer and basic monomer. It also demonstrates that at very low concentration of the basic monomer (about 12 % w/w) this copolymer exhibits unusual dissolution behavior. Contrary to the solubility behavior of Eudragit E polymers, these polymers are soluble over a wide pH range. These polymers are copolymers of lactide macromer and DMAEMA. Because of their unusual dissolution behavior, they can be used as excipients in pharmaceutical drug delivery systems. The lactide macromer used in the present invention is a low molecular weight moiety and the copolymer shows unexpected dissolution properties which are very different than copolymers of DMAEMA reported in the literature.

Objectives of the invention

The main objectivies of the present invention is to provide a macromer based novel copolymer comprising low molecular weight lactide macromer and a basic monomer Another objectivie of the present invention is to provide a process for the preparation of macromer based novel copolymer by copolymerization of acrylate or methacrylate ester of oligomeric lactide with basic DMAEMA monomer.

Summary of the invention

The present invention relates to a copolymer which is synthesized using oligomeric lactide macromer and basic monomer and method of preparation thereof The said copolymer has a very low concentration of the basic monomer (about 12 % w/w) and exhibit unusual behavior . The said copolymers are soluble over a wide PH range and having application in pharmaceutical drug delivery system.

Detailed description of the invention

Accordingly the present invention provides a novel oligomeric lactide macromer based copolymer having the general formula

[A ( _X ) B _(y) ] n wherein A is an oligomeric lactide macromer having a degree of polymerization in the range of 2 to 10, B is a basic monomer, x varies from 1 to 4, y varies from 3 to 7 and n varies from 1 to 5. In an embodiment of the present invention the copolymer prepared is soluble in an aqueous medium over a pH rage of 1.8 to 7.4.

In another embodiment of the present invention the copolymer prepared has a molecular weight in the range of 1000 to 5000.

In yet another embodiment the content of oligomeric lactide macromer present is in the range of 25 to 90 % w/w of the copolymer.

In yet another embodiment the content of basic monomer present is in the range of 10 to 75 % w/w of the copolymer.

In yet another embodiment the macromer A used is an acrylic or methacrylic acid ester selected from oligo ( lactide) acrylate and oligo (lactide) methacrylate .

In yet another embodiment macromer A used has a molecular weight in the range of 500 to 1000.

In yet another embodiment the basic monomer B used is selected from the group comprising of dimethyl amino ethyl methacrylate, dimethyl amino ethyl acrylate, diethyl amino ethyl methacrylate, diethyl amino ethyl acrylate, piperidine ethyl methacrylate and 2 tert-butyl amino ethyl methacrylate. The present invention further provides a process for the preparation of a novel oligomeric lactide macromer based copolymer having the general formula

[A ( _X) B ₍ y ₎ ] _n

wherein A is an oligomeric lactide macromer having a degree of polymerization in the range of 2 to 10, B is a basic monomer, x varies from 1 to 4, y varies from 3 to 7 and n varies from 1 to 5, which comprises a)preparing a solution of a macromer A and a basic monomer B in a molar ratio in the range of 1 to 9 and 9 to 1 with an organic solvent, b)adding a, free radical initiator to above said solution and heating it to a temperature ranging between 50 to 75 ⁰ C ₁ for a period of 16 to 24 hours where by a reaction mixture is obtained, c)concentrating the above said reaction mixture by removing solvent, at a reduced pressure and d) precipitating the resultant reaction mixture in water to recover the desired copolymer.

In yet another embodiment organic solvent of step( a) is in the range 5 to 50 ml. In yet another embodiment free radical initiator of step( b) is in the ratio

1:10 of free radical initiator to solution of step (a)

In yet another embodiment of the present invention the macromer A used is an acrylic or methacrylic acid ester selected from the group comprising of oligo ( lactide) acrylate and oligo (lactide) methacrylate In yet another embodiment of the present invention the the macromer A used has a molecular weight in the range of 500 to 1000.

In yet another embodiment the basic monomer B used is selected from the group comprising of dimethyl amino ethyl methacrylate, dimethyl amino ethyl acrylate, diethyl amino ethyl methacrylate, diethyl amino ethyl acrylate, piperidine ethyl methacrylate and 2 tert-butyl amino ethyl methacrylate. In yet another embodiment the organic solvent used is selected from the group comprising of chlorinated hydrocarbon, alcohol, ester, ketone, formamide, tetrahydrofuran, dioxane and dimethyl sulfoxide.

In yet another embodiment the free radical initiator used is an azocompound selected from the group comprising of azo-bis-cyano-valeric acid, azo-bis-biphenyl methane, azo-bis- methyl isobutyrate and azo-bis- isobutyronitrile.

In yet another embodiment the free radical initiator used is selected from the group comprising of peroxide, hydroperoxide, peracid and perester.

In yet another embodiment the copolymer obtained is soluble in an aqueous medium over a pH rage of 1.8 to 7.4.

In yet another embodiment the copolymer obtained has a molecular weight in the range of 1000 to 5000.

In yet another embodiment the copolymer obtained has a content of oligomeric lactide macromer in the range of 25 to 90 % w/w of the copolymer. In still another embodiment the copolymer obtained has a content of basic monomer B in the range of 10 to 75 % w/w of the copolymer.

High-molecular-weight linear polyesters like polylactides, polyglycolides and their copolymers are widely used for medical applications such as surgical sutures, drug delivery devices, tissue supports, and implants for internal bone fixation. But less attention has been paid to lactide oligomers as they lack mechanical and thermal properties required for above applications. These low melt oligolactides can be looked upon as a new class of biodegradable materials useful in injectable drug delivery systems, implant coatings or soft tissue augmentations. This invention relates to the synthesis of novel lactide copolymers from low molecular weight lactide macromer and basic monomer. Copolymer

compositions of the present invention can be obtained by varying two principal factors in the process.

1. The molecular weight of lactide macromer.

2. Composition of macromer and basic monomer in feed during polymerization In the present invention the lactide macromer (A) is synthesized from oligo (lactide) diol by condensation reaction with acryloyl chloride or methacryloyl chloride using triethyl amine as base.( Haris, Parvez I.; Vogt, S.; Berger, S.; Wilke, I.; Larcher, Y.; Weisser, J.; Schnabelrauch, M. Bio-Medical Materials and Engineering 15(1, 2), (2005), pp.73-85) This macromer can be also synthesized by coupling oligomeric diol with acrylic acid or methacrylic acid using Dicyclohexyl carbodiimide. Oligomeric lactide diol is an oligomeric lactide having terminal hydroxyl groups which is synthesized from Lactide and 1 , 4 Butanediol by ring opening melt polymerization. (Kari Hiltunen, Mika Harkonen, Jukka Seppala, Taito Vaananen Macromolecules 29, 1996, pp.8677-8682). This oligo (lactide) diol is then , dissolved in tetrahydrofuran to which acryloyl chloride is added drop by drop under Nitrogen atmosphere. The product is recovered by removing salt by filtration. Then macromer is precipitated in non-solvent like water and dried at room temperature. This macromer is then used for copolymerization with a basic monomer.

The solution polymerization technique is used for polymerization of oligomeric lactide macromer with basic monomer. In solution polymerization, the macromer and basic monomer are dissolved in the solvent. Azo-bis- isobutyronitrile is used as initiator for polymerization. Reaction mixture is purged with Nitrogen and the polymerization carried out under inert atmosphere. After stipulated reaction time, solvent is evaporated under reduced pressure and the polymer is precipitated from the solution, by precipitating in nonsolvents like water, petroleum ether or diethyl ether. The polymer is then dried under vacuum. The dissolution behavior of the polymers synthesized was studied by exposing the polymers to buffer solutions of different range of 1.8, 4.8, 6.8 and

7.4 pH. 50 mg dry polymer of each composition was put in different test tube in

5 ml buffer of different pH.

Dissolution behavior of the polymers is recorded in table 1 and 2.

The advantage of the disclosed invention is thus attained in an economical, practical and facile manner. While preferred embodiments and example configurations have been shown and described, it is to be understood that various further modifications and additional configuration will be apparent to those skilled in the art. It is intended that the specific embodiments and configuration herein disclosed are illustrative of the preferred to best mode of practicing the invention and should not be interpreted as limitation of the scope of the invention.

Examplei

This example provides for the preparation of copolymer of poly (Lactide) acrylate with Dimethyl amino ethyl methacrylate in which DMAEMA content is 14 % w/w

1.0 g (1.42 x10 ^'3 moles) poly (Lactide) acrylate was dissolved in 13 ml Dimethyl formamide to which 0.240 ml (1.42 x 10 ^"3 moles) Dimethyl amino ethyl methacrylate was added. The initiator azo bis lsobutyronitrile 9.32 mg (5.68x 10 ^"5 moles) was added to it. This reaction mixture was stirred well and nitrogen was purged through it for 10 minutes. This was heated for 24 hours at 65 ⁰ C in a water bath. After predetermined reaction time, solvent was removed under reduced pressure and the polymer was precipitated in water. Polymer was dried under vacuum and characterized by NMR and VPO. NMR of the copolymer was recorded in deuterated chloroform. Signals observed in the spectrum which were used for composition analysis were (2 protons of two -CH of lactide at 5.16 with 2 protons of -OCH ₂ of DMAEMA at 4.2)

Example 2 This example provides for the preparation of copolymer of poly (Lactide) methacrylate with Dimethyl amino ethyl methacrylate in which DMAEMA content is 72 % w/w

1.0 g (1.40x10 ^"3 moles) poly (Lactide) methacrylate was dissolved in 10 ml Dimethyl formamide to which 1.99 g (0.0126 moles) Dimethyl amino ethyl methacrylate was added. The initiator azo bis lsobutyronitrile 45 mg (2.8 x 10 ^"4 moles) was added to it. This reaction mixture was stirred well and nitrogen was purged through it for 10 minutes. This was heated for 18 hours at 60 ⁰ C in a water bath. After stipulated reaction time, solvent was removed under reduced pressure and the polymer was precipitated in water. Polymer was dried under vacuum and characterized by NMR and VPO.

Example 3

This example provides for the preparation of copolymer of poly (Lactide) acrylate with Dimethyl amino ethyl methacrylate in which poly (lactide) acrylate molecular weight was 700. 1.0 g (1.42 x10 ^'3 moles) poly (Lactide) acrylate was dissolved in 13 ml Dimethyl formamide to which 0.957 ml (5.68 x 10 ^"3 moles) Dimethyl amino ethyl methacrylate was added. The initiator azo bis lsobutyronitrile 23 mg (1.42x 10 ^"4 moles) was added to it. This reaction mixture was stirred well and nitrogen was purged through it for 10 minutes. This was heated for 20 hours at 70 ⁰ C in a water bath. After predetermined reaction time, solvent was removed under reduced pressure and polymer was precipitated in water. Polymer was dried under vacuum and characterized by NMR andVPO.

Example 4

This example provides for the preparation of copolymer of poly (Lactide) methacrylate with Dimethyl amino ethyl methacrylate in which poly (lactide) methacrylate molecular weight was 800.

1.0 g (1.25x10 ^"3 moles) poly (Lactide) methacrylate was dissolved in 10 ml Dimethyl formamide to which 0.210 ml (1.25x10 ^"3 moles) Dimethyl amino ethyl methacrylate was added. The initiator azo bis lsobutyronitrile 8.21 mg (5.Ox 10 ^{' 5} moles) was added to it. This reaction mixture was stirred well and nitrogen was purged through it for 10 minutes. This was heated for 22 hours at 55 ⁰ C in a water bath. After stipulated reaction time, solvent was removed under reduced

pressure and the polymer was precipitated in water. Polymer was dried under vacuum and characterized by NMR and VPO.

Vapour pressure osmometer (VPO) was used for molecular weight measurement of all these DMAEMA copolymers was from KNAUER K 7000 series. Solvent used for measurement was chloroform (HPLC grade).

Table 1 Dissolution behavior of polymers in buffers

Composition- oligo (Lactide) acrylate: DMAEMA

Table 2 Dissolution behavior of polymers in buffers

Composition: oligo (lactide) methacrylate: DMAEMA

_g

Advantages of the present invention are:

1. It exhibits unusual dissolution behavior and are soluble in aqueous medium over a wide pH range of 1.8 to 7.4.

2. Because of their unusual dissolution behavior, they can be used as excipients in pharmaceutical drug delivery systems.

3. The lactide macromer used in the present invention is a low molecular weight moiety.