Patent Publication Number: US-2021186867-A1

Title: Drug delivery system for the delivery of antiviral agents

Description:
CROSS REFERENCE TO RELATED APPLICATIONS 
     This application is a continuation of application Ser. No. 16/310,511, filed Dec. 17, 2018, which a U.S. National Phase application under 35 U.S.C. § 371 of PCT Application No. PCT/US2017/037583 filed Jun. 15, 2017, which claims priority from U.S. Ser. No. 62/352,208 filed Jun. 20, 2016. 
    
    
     BACKGROUND OF THE INVENTION 
     The development of highly active antiretroviral therapy (HAART) in the mid 1990&#39;s transformed the clinical care of human immunodeficiency virus (HIV) type infection. HAART regimens have proven to be highly effective treatments, significantly decreasing HIV viral load in HIV-infected patients, thereby slowing the evolution of the illness and reducing HIV-related morbidity and mortality. Yet, the treatment success of HAART is directly related to adherence to the regimen by the patient. Unless appropriate levels of the antiretroviral drug combinations are maintained in the blood, viral mutations will develop, leading to therapy resistance and cross-resistances to molecules of the same therapeutic class, thus placing the long-term efficacy of treatments at risk. Various clinical studies have shown a decline in treatment effectiveness with relatively small lapses in adherence. A study by Musiime found that 81% of patients with more than 95% adherence demonstrated viral suppression, while only 50% of patients who were 80-90% adherent were successful. See, Musiime, S., et al., Adherence to Highly Active Antiretroviral Treatment in HIV-Infected Rwandan Women.  PLOS  one 2011, 6, (11), 1-6. Remarkably, only 6% of patients that were less than 70% adherent showed improvements in viral markers. Thus, low adherence is a leading cause of therapeutic failure in treatment of HIV-1 infection. 
     Nonetheless, adherence rates to the HAART regimens continue to be far from optimal. Various characteristics of HAART make adherence particularly difficult. Therapeutic regimens are complex, requiring multiple drugs to be taken daily, often at different times of the day, and many with strict requirements on food intake. Many HAART medications also have unpleasant side effects, including nausea, diarrhea, headache, and peripheral neuropathy. Social and psychological factors can also negatively impact adherence. Patients report that forgetfulness, lifestyle factors, including fear of being identified as HIV-positive, and therapy fatigue over life-long duration of treatment all contribute to adherence lapses. 
     New HIV treatment interventions aim to improve adherence by reducing the complexity of treatments, the frequency of the dosages, and/or the side effects of the medications. Long-acting injectable (LAI) drug formulations that permit less frequent dosing, on the order of a month or longer, are an increasingly attractive option to address adherence challenges. However, the majority of approved and investigational antiretroviral agents are not well suited for reformulation as long-acting injectable products. In large part, this is due to suboptimal physicochemical properties limiting their formulation as conventional drug suspensions, as well as insufficient antiviral potency resulting in high monthly dosing requirements. Even for cabotegravir or rilpivirine, two drugs being studied as long-acting injectible formulations, large injection volumes and multiple injections are required to achieve pharmacokinetic profiles supportive of monthly dosing. See, e.g., Spreen, W. R., et al., Long-acting injectable antiretrovirals for HIV treatment and prevention.  Current Opinion in Hiv and Aids  2013, 8, (6), 565-571; Rajoli, R. K. R., et al., Physiologically Based Pharmacokinetic Modelling to Inform Development of Intramuscular Long-Acting Nanoformulations for HIV.  Clinical Pharmacokinetics  2015, 54, (6), 639-650; Baert, L., et al., Development of a long-acting injectable formulation with nanoparticles of rilpivirine (TMC278) for HIV treatment.  European Journal of Pharmaceutics and Biopharmaceutics  2009, 72, (3), 502-508; Van&#39;t Klooster, G., et al., Pharmacokinetics and Disposition of Rilpivirine (TMC278) Nanosuspension as a Long-Acting Injectable Antiretroviral Formulation.  Antimicrobial Agents and Chemotherapy  2010, 54, (5), 2042-2050. Thus, novel formulation approaches capable of delivering extended-duration pharmacokinetic characteristics for molecules of diverse physicochemical properties at practical injection volumes and with a limited number of injections are highly desirable. 
     SUMMARY OF THE INVENTION 
     This invention relates to novel implant drug delivery systems for long-acting delivery of antiviral drugs. These compositions are useful for the treatment or prevention of human immunodeficiency virus (HIV) infection. 
    
    
     DETAILED DESCRIPTION OF THE INVENTION 
     This invention relates to novel implant drug delivery systems for long-acting delivery of antiviral drugs. The novel implant drug delivery systems comprise a polymer and an antiviral agent. These implant drug delivery systems are useful for the treatment or prevention of human immunodeficiency virus (HIV) infection. The invention further relates to methods of treating and preventing HIV infection with the novel implant drug delivery systems described herein. 
     The novel implant delivery systems of the invention comprise a biocompatible bioerodible polymer to generate monolithic matrices with dispersed or dissolved drug. The chemical properties of the polymer matrices are tuned to achieve a range of drug release characteristics, offering the opportunity to extend duration of dosing. In an embodiment of the invention, the novel implant delivery systems are compatible with molecules having a broad spectrum of physicochemical properties, including those of high aqueous solubility or amorphous phases which are unsuitable to formulation as solid drug suspensions. 
     Specifically, this invention relates to novel implant drug delivery systems comprising a biocompatible bioerodible polymer and 4′-ethynyl-2-fluoro-2′-deoxyadenosine wherein said implant drug delivery system is implanted subdermally and 4′-ethynyl-2-fluoro-2′-deoxyadenosine is continually released in vivo at a rate resulting in a plasma concentration between 0.01 ng/mL and 3000.0 ng/mL. These implant delivery systems are desired and useful for prophylaxis and/or treatment of HIV infection from both compliance and convenience standpoints. 
     As used herein, the term “biocompatible bioerodible polymer” refers to polymeric materials that include hydrolytically labile linkages which undergo cleavage at physiological conditions. The broken down-products are non-toxic and either excreted in the urine or incorporated into the Krebs cycle and used for energy. The polymer is generally hydrophobic so that it retains its integrity for a suitable period of time when placed in an aqueous environment, such as the body of a mammal, and is stable enough to be stored for an extended period before use. Bioerodible polymers remain intact in vivo for extended periods of time, typically weeks, months or years. Drug molecules encapsulated in the polymer are released over time via diffusion through channels and pores in a sustained manner. The release rate can be altered by modifying the identity of the polymer (monomeric units, molecular weight, end group, etc.) thereby modifying the degradation kinetics, percent drug loading, porosity of the polymer, structure of the implantable device, or hydrophobicity of the polymer, or by adding a hydrophobic coating to the exterior of the implantable device. 
     Accordingly, any polymer that can be readily cleared or eliminated by the body can be used to manufacture the implant drug delivery systems of the instant invention that comprise a biocompatible bioerodible polymer. The term “polymer” can also include copolymers. Biocompatible bioerodible polymers of the instant invention include, but are not limited to, poly(DL-lactide) (“PLA”), poly(caprolactone) (“PCL”), poly(lactide-co-glycolide), poly(lactide), poly(glycolide) (“PLG”), poly(ortho esters), poly(dioxanone), poly(alkylcyanoacrylates) and combinations thereof. In a class of the invention, the biocompatible bioerodible polymer is selected from the group consisting of poly(DL-lactide) and poly(caprolactone). 
     In a class of the invention, the biocompatible bioerodible polymer is selected from the group consisting of poly(DL-lactide), poly(L-lactide), and poly(caprolactone), all of which can have an acid or an ester end group. In a subclass of the invention, the biocompatible bioerodible polymer is poly(DL-lactide). In another subclass of the invention, the biocompatible bioerodible polymer is poly(caprolactone). 
     As used herein, the term “diffusional barrier” refers to a coating that is permeable to the drug and is placed over at least a portion of the device to further regulate the rate of release. For example, a coating of biocompatible bioerodible polymeric material, e.g., poly(DL-lactide), or a coating of a biocompatible bioerodible polymeric material with a lower drug loading than the remainder of the implant delivery system, may be used. The diffusional barrier may be formed, for example, by coextrusion with the device. 
     Suitable diffusional barriers of the instant invention include, but are not limited to, poly(DL-lactide) (“PLA”), poly(caprolactone) (“PCL”), poly(lactide-co-glycolide), poly(lactide), poly(glycolide), poly(ortho esters), poly(dioxanone), poly(alkylcyanoacrylates) and combinations thereof. In a class of the invention, the diffusional barrier is selected from the group consisting of poly(DL-lactide) and poly(caprolactone). 
     In an embodiment of the invention, the diffusion barrier contains an antiviral drug. In a class of the embodiment, the diffusion barrier comprises 4′-ethynyl-2-fluoro-2′-deoxyadenosine. 
     As used herein, the term “dispersed or dissolved in the biocompatible bioerodible polymer” refers to the drug and polymer being mixed and then hot-melt extruded. 
     As used herein, the term “continually released” refers to the drug being released from the biocompatible bioerodible polymer at continuous rates for extended periods of time. The implant drug delivery systems of the instant invention generally exhibit 1 st  order release kinetics for the drug in vivo, sometimes with an initial burst. Polymer degradation modifies the dissolution and diffusion of the drug from the polymer matrix. This typically results in an increased drug elution rate that may deviate from 1 st  order kinetics, and is a function of the polymer degradation rate. 
     Optionally, the novel implant delivery systems of the instant invention can further comprise a radiopaque component. The radiopaque component will cause the implant to be X-ray visible. The radiopaque component can be any such element known in the art, such as barium sulfate, titanium dioxide, bismuth oxide, tantalum, tungsten or platinum. In a specific embodiment, the radiopaque component is barium sulfate. 
     In one embodiment, the radiopaque material is about 1% to 30% by weight. In another embodiment, the radiopaque material is about 1% to 20% by weight. In another embodiment, the radiopaque material is about 4% to 25% by weight. In further embodiment, the radiopaque material is about 6% to 20% by weight. In another embodiment, the radiopaque material is about 4% to 15% by weight. In another embodiment, the radiopaque material is about 8% to 15% by weight. 
     The radiopaque material does not affect the release of 4′-ethynyl-2-fluoro-2′-deoxyadenosine from the implant. 
     The novel implant delivery systems of the invention comprise antiviral agents. Suitable antiviral agents include anti-HIV agents. In an embodiment of the invention, the antiviral agent is administered as a monotherapy. In another embodiment of the invention, two or more antiviral agents are administered in combination. 
     An “anti-HIV agent” is any agent which is directly or indirectly effective in the inhibition of HIV reverse transcriptase or another enzyme required for HIV replication or infection, or prophylaxis of HIV infection, and/or the treatment, prophylaxis or delay in the onset or progression of AIDS. It is understood that an anti-HIV agent is effective in treating, preventing, or delaying the onset or progression of HIV infection or AIDS and/or diseases or conditions arising therefrom or associated therewith. Suitable anti-viral agents for use in implant drug delivery systems described herein include, for example, those listed in Table A as follows: 
     
       
         
           
               
             
               
                 TABLE A 
               
             
            
               
                   
               
               
                 Antiviral Agents for Preventing HIV infection or AIDS 
               
            
           
           
               
               
            
               
                 Name 
                 Type 
               
               
                   
               
               
                 abacavir, ABC, Ziagen ® 
                 nRTI 
               
               
                 abacavir + lamivudine, Epzicom ® 
                 nRTI 
               
               
                 abacavir + lamivudine + zidovudine, Trizivir ® 
                 nRTI 
               
               
                 amprenavir, Agenerase ® 
                 PI 
               
               
                 atazanavir, Reyataz ® 
                 PI 
               
               
                 AZT, zidovudine, azidothymidine, Retrovir ® 
                 nRTI 
               
               
                 Capravirine 
                 nnRTI 
               
               
                 darunavir, Prezista ® 
                 PI 
               
               
                 ddC, zalcitabine, dideoxycytidine, Hivid ® 
                 nRTI 
               
               
                 ddI, didanosine, dideoxyinosine, Videx ® 
                 nRTI 
               
               
                 ddI (enteric coated), Videx EC ® 
                 nRTI 
               
               
                 delavirdine, DLV, Rescriptor ® 
                 nnRTI 
               
               
                 doravirine 
                 nnRTI 
               
               
                 efavirenz, EFV, Sustiva ®, Stocrin ® 
                 nnRTI 
               
               
                 efavirenz + emtricitabine + tenofovir DF, Atripla ® 
                 nnRTI + nRTI 
               
               
                 EFdA (4′-ethynyl-2-fluoro-2′-deoxyadenosine) 
                 nRTI 
               
               
                 emtricitabine, FTC, Emtriva ® 
                 nRTI 
               
               
                 emtricitabine + tenofovir DF, Truvada ® 
                 nRTI 
               
               
                 emvirine, Coactinon ® 
                 nnRTI 
               
               
                 enfuvirtide, Fuzeon ® 
                 FI 
               
               
                 enteric coated didanosine, Videx EC ® 
                 nRTI 
               
               
                 etravirine, TMC-125 
                 nnRTI 
               
               
                 fosamprenavir calcium, Lexiva ® 
                 PI 
               
               
                 indinavir, Crixivan ® 
                 PI 
               
               
                 lamivudine, 3TC, Epivir ® 
                 nRTI 
               
               
                 lamivudine + zidovudine, Combivir ® 
                 nRTI 
               
               
                 Lopinavir 
                 PI 
               
               
                 lopinavir + ritonavir, Kaletra ® 
                 PI 
               
               
                 maraviroc, Selzentry ® 
                 EI 
               
               
                 nelfinavir, Viracept ® 
                 PI 
               
               
                 nevirapine, NVP, Viramune ® 
                 nnRTI 
               
               
                 PPL-100 (also known as PL-462) (Ambrilia) 
                 PI 
               
               
                 raltegravir, Isentress ™ 
                 InI 
               
               
                 (S)-2-(3-chloro-4-fluorobenzyl)-8-ethyl-10-hydroxy- 
                 InI 
               
               
                 N,6-dimethyl-1,9-dioxo-1,2,6,7,8,9- 
                   
               
               
                 hexahydropyrazino[1′,2′:1,5]pyrrolo[2,3-d]pyridazine- 
                   
               
               
                 4-carboxamide (MK-2048) 
                   
               
               
                 ritonavir, Norvir ® 
                 PI 
               
               
                 saquinavir, Invirase ®, Fortovase ® 
                 PI 
               
               
                 stavudine, d4T, didehydrodeoxythymidine, Zerit ® 
                 nRTI 
               
               
                 tenofovir DF (DF = disoproxil fumarate), TDF, 
                 nRTI 
               
               
                 Viread ® 
                   
               
               
                 Tenofovir, hexadecyloxypropyl (CMX-157) 
                 nRTI 
               
               
                 tipranavir, Aptivus ® 
                 PI 
               
               
                 Vicriviroc 
                 EI 
               
               
                   
               
               
                 EI = entry inhibitor; 
               
               
                 FI = fusion inhibitor; 
               
               
                 InI = integrase inhibitor; 
               
               
                 PI = protease inhibitor; 
               
               
                 nRTI = nucleoside reverse transcriptase inhibitor; 
               
               
                 nnRTI = non-nucleoside reverse transcriptase inhibitor. 
               
            
           
         
       
     
     Some of the drugs listed in the table can be used in a salt form; e.g., abacavir sulfate, delavirdine mesylate, indinavir sulfate, atazanavir sulfate, nelfinavir mesylate, saquinavir mesylate. 
     In certain embodiments the antiviral agents in the implant drug delivery systems described herein are employed in their conventional dosage ranges and regimens as reported in the art, including, for example, the dosages described in editions of the  Physicians&#39; Desk Reference , such as the 70th edition (2016) and earlier editions. In other embodiments, the antiviral agents in the implant drug delivery systems described herein are employed in lower than their conventional dosage ranges. 
     In an embodiment of the invention, the antiviral agent can be an entry inhibitor; fusion inhibitor; integrase inhibitor; protease inhibitor; nucleoside reverse transcriptase inhibitor; or non-nucleoside reverse transcriptase inhibitor. In a class of the invention, the antiviral agent is a nucleoside reverse transcription inhibitor. 
     In an embodiment of the invention, the antiviral agent is a nucleoside reverse transciptase inhibitor (NRTI). In a class of the invention, the NRTI is 4′-ethynyl-2-fluoro-2′-deoxyadenosine. 
     4′-ethynyl-2-fluoro-2′-deoxyadenosine is also known as EFdA, and has the following chemical structure: 
     
       
         
         
             
             
         
       
     
     Production of and the ability of 4′-ethynyl-2-fluoro-2′-deoxyadenosine to inhibit HIV reverse transcriptase are described in PCT International Application WO2005090349, published on Sep. 29, 2005, and US Patent Application Publication No. 2005/0215512, published on Sep. 29, 2005, both to Yamasa Corporation which are hereby incorporated by reference in their entirety. 
     In an embodiment of the implant drug delivery system described herein, the antiviral agent is present in the biocompatible bioerodible polymer at about 0.10%-80% by weight of drug loading. In other embodiments, the antiviral agent is present in the biocompatible bioerodible polymer at about 20%-60% by weight, at about 30%-65% by weight, at about 40%-60% by weight or at about 40%-45% by weight of drug loading. In a class of the embodiment of the implant drug delivery system described herein, 4′-ethynyl-2-fluoro-2′-deoxyadenosine is present in the biocompatible bioerodible polymer at about 0.10%-80% by weight of drug loading. In a subclass of the embodiment of the implant drug delivery system described herein, 4′-ethynyl-2-fluoro-2′-deoxyadenosine is present in the biocompatible bioerodible polymer at about 20%-60% by weight of drug loading. In a further subclass of the embodiment of the implant drug delivery system described herein, 4′-ethynyl-2-fluoro-2′-deoxyadenosine is present in the biocompatible bioerodible polymer at about 30%-65% by weight of drug loading. In a further subclass of the embodiment of the implant drug delivery system described herein, 4′-ethynyl-2-fluoro-2′-deoxyadenosine is present in the biocompatible bioerodible polymer at about 40%-60% by weight of drug loading. In a further subclass of the embodiment of the implant drug delivery system described herein, 4′-ethynyl-2-fluoro-2′-deoxyadenosine is present in the biocompatible bioerodible polymer at about 40%-45% by weight of drug loading. In an example of the embodiment of the implant drug delivery system described herein, 4′-ethynyl-2-fluoro-2′-deoxyadenosine is present in the biocompatible bioerodible polymer at 40% by weight of drug loading. In another example of the embodiment of the implant drug delivery system described herein, 4′-ethynyl-2-fluoro-2′-deoxyadenosine is present in the biocompatible bioerodible polymer at 45% by weight of drug loading. In another example of the embodiment of the implant drug delivery system described herein, 4′-ethynyl-2-fluoro-2′-deoxyadenosine is present in the biocompatible bioerodible polymer at 50% by weight of drug loading. In another example of the embodiment of the implant drug delivery system described herein, 4′-ethynyl-2-fluoro-2′-deoxyadenosine is present in the biocompatible bioerodible polymer at 60% by weight of drug loading. In another example of the embodiment of the implant drug delivery system described herein, 4′-ethynyl-2-fluoro-2′-deoxyadenosine is present in the biocompatible bioerodible polymer at 80% by weight of drug loading. 
     The implant drug delivery systems of the instant invention may be produced using an extrusion process, wherein ground biocompatible, bioerodible polymer is blended with the antiviral agent, melted and extruded into rod-shaped structures. Rods are cut into individual implantable devices of the desired length, packaged and sterilized prior to use. Other methods for encapsulating therapeutic compounds in implantable polymeric, bioerodible matrices are known to those of skill in the art. Such methods include solvent casting (see U.S. Pat. Nos. 4,883,666, 5,114,719 and 5,601835). One of skill in the art would be able to readily determine an appropriate method of preparing such an implant drug delivery system, depending on the shape, size, drug loading, and release kinetics desired for a particular type of patient or clinical application. 
     The size and shape of the implant drug delivery systems may be modified to achieve a desired overall dosage. The implant drug delivery systems of the instant invention are often about 0.5 cm to about 10 cm in length. In an embodiment of the invention, the implant drug delivery systems are about 1.5 cm to about 5 cm in length. In a class of the embodiment, the implant drug delivery systems are about 2 cm to about 5 cm in length. In a subclass of the embodiment, the implant drug delivery systems are about 2 cm to about 4 cm in length. The implant drug delivery systems of the instant invention are often about 0.5 mm to about 7 mm in diameter. In an embodiment of the invention, the implant drug delivery systems are about 1.5 mm to about 5 mm in diameter. In a class of the embodiment, the implant drug delivery systems are about 2 mm to about 5 mm in diameter. In a subclass of the embodiment, the implant drug delivery systems are about 2 mm to about 4 mm in diameter. 
     The implant drug delivery systems described herein are capable of releasing 4′-ethynyl-2-fluoro-2′-deoxyadenosine over a period of 21 days, 28 days, 31 days, 4 weeks, 6 weeks, 8 weeks, 12 weeks, one month, two months, three months, four months, five months, six months, seven months, eight months, nine months, ten months, eleven months, twelve months, eighteen months, twenty-four months or thirty-six months at an average rate of between 0.01-5 mg per day. In an embodiment of the invention, the 4′-ethynyl-2-fluoro-2′-deoxyadenosine is released at therapeutic concentrations for a duration from between three months and thirty-six months. In a class of the embodiment, the 4′-ethynyl-2-fluoro-2′-deoxyadenosine is released at therapeutic concentrations for a duration from between six months and twelve months. 
     One or more implants can be used to achieve the desired therapeutic dose. In an embodiment of the invention, one or more implants can be used to achieve the therapeutic dose for durations of up to 1 year. In another embodiment of the invention, one or more implants can be used to achieve the therapeutic dose for durations of up to 2 years. 
     The implant drug delivery systems described herein are capable of releasing 4′-ethynyl-2-fluoro-2′-deoxyadenosine resulting in a plasma concentration of between 0.02-300 ng/mL per day. In an embodiment of the invention, the implant drug delivery systems described herein are capable of releasing 4′-ethynyl-2-fluoro-2′-deoxyadenosine resulting in a plasma concentration of between 0.02-30.0 ng/mL per day. In a class of the embodiment, the implant drug delivery systems described herein are capable of releasing 4′-ethynyl-2-fluoro-2′-deoxyadenosine resulting in a plasma concentration of between 0.02-15 ng/mL per day. In a class of the embodiment, the implant drug delivery systems described herein are capable of releasing 4′-ethynyl-2-fluoro-2′-deoxyadenosine resulting in a plasma concentration of between 0.02-8 ng/mL per day. In a subclass of the embodiment, the implant drug delivery systems described herein are capable of releasing 4′-ethynyl-2-fluoro-2′-deoxyadenosine resulting in a plasma concentration of between 0.1-1.0 ng/mL per day. 
     The following examples are given for the purpose of illustrating the present invention and shall not be construed as being limitations on the scope of the invention. 
     Example 1 
     Preparation and In Vitro Release of Implant Drug Delivery Systems Containing 40-80 Wt % 4′-Ethynyl-2-Fluoro-2′-Deoxyadenosine 
     Implantable devices were prepared using an extrusion process. The first step involved mixing the dry, micronized powders of the active compound (4′-ethynyl-2-fluoro-2′-deoxyadenosine) and the cryomilled PCL (Evonik RESOMER® Select 100 CL 7.5E) or PLA (Evonik RESOMER® Select 100 DL 8A) using a Turbula T2F mixer. Drug and polymer blends were prepared at 40-80 wt % drug load. The drug and polymer blends were hot-melt extruded using a twin screw extruder through a 3 mm diameter die and pulled to a diameter of approximately 1.9-2.3 mm. The screws contained predominately conveying elements with a single 90° mixing section. The 1St zone where the drug-polymer blends were introduced was water-cooled and maintained at room temperature. The temperature for zones 2-4 was 100° C. for PLA and 75° C. for PCL. Extruded fibers with diameters between 1.9-2.3 mm were cut to a length of approximately 40 mm. 
     The in vitro release rate of 4′-ethynyl-2-fluoro-2′-deoxyadenosine was determined by incubating the implants segments, approximately 1 cm in length, in a glass vial containing phosphate buffered saline (PBS) at 37° C., and 50 rpm shaking in an Innova 42 incubator. The volume of PBS was sufficient to maintain sink conditions. Sink conditions are defined as the drug concentration maintained at or below ⅓ of the maximum solubility (drug concentration &lt;0.45 mg/mL in PBS at 37° C.). Samples were removed (0.5 mL) at selected time points, and centrifuged at 20,800×g for 8 min. The supernatant was removed (0.4 mL), diluted 4-fold, and vortexed. Samples were assayed by HPLC (Agilent 1100 series). Analysis of a 6 μL volume was performed at 240 nm with a Supelco Ascentis® Express C18 column (100×4.6 mm, 2.7 μm). The mobile phase was 0.1% H 3 PO 4  and 50:50 ACN:MeOH (83:17 v/v) at a flow rate of 1.5 mL/min (40° C.). 
     To determine degradation of 4′-ethynyl-2-fluoro-2′-deoxyadenosine by HPLC, a 6 μL volume was injected onto an Agilent Zorbax SB-Aq column (150×4.6 mm, 3.5 μm). The mobile phase was 0.1% H 3 PO 4  and 50:50 ACN:MeOH with a flow rate of 1.0 mL/min (40° C.). The mobile phase gradient is shown in the table below. 
     
       
         
           
               
             
               
                 TABLE 1 
               
             
            
               
                   
               
               
                 4′-ethynyl-2-fluoro-2′-deoxyadenosine chemical stability 
               
               
                 HPLC method details 
               
            
           
           
               
               
               
            
               
                   
                 Time (min) 
                 0.1% H 3 PO 4   
               
               
                   
               
            
           
           
               
               
               
            
               
                   
                 0.0 
                 98 
               
               
                   
                 10.0 
                 95 
               
               
                   
                 12.0 
                 90 
               
               
                   
                 14.0 
                 10 
               
               
                   
                 14.1 
                 98 
               
               
                   
                 20.0 
                 98 
               
               
                   
               
            
           
         
       
     
     All samples were calibrated to 0.5 mg/mL standard solutions of 4′-ethynyl-2-fluoro-2′-deoxyadenosine in 50:50 MeOH:H 2 O. 
     
       
         
           
               
             
               
                 TABLE 2 
               
             
            
               
                   
               
               
                 4′-ethynyl-2-fluoro-2′-deoxyadenosine (EFdA) in vitro release from 40 wt %, 50 wt %, 
               
               
                 60 wt %, and 80 wt % 4′-ethynyl-2-fluoro-2′-deoxyadenosine in poly(DL-lactide) 
               
               
                 (PLA) implants at sink conditions; reported as a % release from total  
               
               
                 [avg = average and std dev = standard deviation] 
               
            
           
           
               
               
               
               
               
            
               
                   
                 40 wt % EFdA + 
                 50 wt % EFdA + 
                 60 wt % EFdA + 
                 80 wt % EFdA + 
               
               
                   
                 60 wt % PLA 
                 50 wt % PLA 
                 40 wt % PLA 
                 20 wt % PLA 
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                 Time 
                   
                 Std. 
                   
                 Std. 
                   
                 Std. 
                   
                 Std. 
               
               
                 (days) 
                 Avg (%) 
                 Dev. 
                 Avg (%) 
                 Dev. 
                 Avg (%) 
                 Dev. 
                 Avg (%) 
                 Dev. 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                 3 
                 4 
                 1 
                 12 
                 2 
                 24 
                 4 
                 39 
                 3 
               
               
                 7 
                 9 
                 2 
                 26 
                 3 
                 48 
                 2 
                 69 
                 13 
               
               
                 14 
                 16 
                 4 
                 42 
                 2 
                 68.7 
                 0.5 
                 91 
                 3 
               
               
                 21 
                 23 
                 6 
                 56 
                 2 
                 85 
                 2 
                 105 
                 3 
               
               
                 27 
                 27 
                 6 
                 63 
                 2 
                 89 
                 1 
                   
                   
               
               
                 35 
                 33 
                 7 
                 72 
                 2 
                 92 
                 1 
                   
                   
               
               
                 42 
                 37 
                 8 
                 78 
                 1 
                 91 
                 1 
                   
                   
               
               
                 49 
                 42 
                 8 
                 83 
                 1 
                 90 
                 1 
                   
                   
               
               
                 63 
                 54 
                 9 
                 94.8 
                 0.3 
                 95 
                 1 
                   
                   
               
               
                 101 
                 80 
                 10 
                   
                   
                   
                   
                   
                   
               
               
                 122 
                 86 
                 8 
                   
                   
                   
                   
                   
                   
               
               
                 136 
                 92 
                 8 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 3 
               
             
            
               
                   
               
               
                 4′-ethynyl-2-fluoro-2′-deoxyadenosine (EFdA) in vitro  
               
               
                 release from 40 wt % and 60 wt % 4′-ethynyl-2-fluoro- 
               
               
                 2′-deoxyadenosine in poly(caprolactone) (PCL) implants  
               
               
                 at sink conditions; reported as a % release from total  
               
               
                 [avg = average and std dev = standard deviation] 
               
            
           
           
               
               
               
            
               
                   
                 40 wt % EFdA + 
                 60 wt % EFdA + 
               
               
                   
                 60 wt % PCL 
                 40 wt % PCL 
               
            
           
           
               
               
               
               
               
            
               
                 Time 
                 Avg  
                 Std. 
                 Avg  
                 Std. 
               
               
                 (days) 
                 (%) 
                 Dev. 
                 (%) 
                 Dev. 
               
               
                   
               
            
           
           
               
               
               
               
               
            
               
                 3 
                 10 
                 1 
                 26 
                 3 
               
               
                 7 
                 15 
                 1 
                 45 
                 5 
               
               
                 14 
                   
                   
                 58 
                 2 
               
               
                 15 
                 21 
                 2 
                   
                   
               
               
                 21 
                   
                   
                 69 
                 2 
               
               
                 23 
                 26 
                 2 
                   
                   
               
               
                 27 
                   
                   
                 75 
                 3 
               
               
                 35 
                 31 
                 2 
                 82 
                 3 
               
               
                 42 
                   
                   
                 86 
                 3 
               
               
                 49 
                   
                   
                 89 
                 2 
               
               
                 63 
                   
                   
                 99 
                 1 
               
               
                 67 
                 47 
                 9 
                   
                   
               
               
                 73 
                 48 
                 4 
                   
                   
               
               
                 86 
                 52 
                 5 
                   
                   
               
               
                 101 
                 55 
                 6 
                   
                   
               
               
                 157 
                 74 
                 7 
                   
                   
               
               
                 178 
                 74 
                 7 
                   
                   
               
               
                 192 
                 75 
                 12 
                   
                   
               
               
                 219 
                 81 
                 7 
                   
                   
               
               
                 231 
                 82 
                 7 
               
               
                   
               
            
           
         
       
     
     Example 2 
     Implant Drug Delivery Systems with Durations of Action of 1 Week to 3 Years 
     Implantable devices were prepared using an extrusion process. The first step involved mixing the dry, micronized powders of the active compound (4′-ethynyl-2-fluoro-2′-deoxyadenosine) and the cryomilled PCL (Evonik RESOMER® Select 100 CL 7.5E) or PLA (Evonik RESOMER® Select 100 DL 8A) using a Turbula T2F mixer. Drug and polymer blends were prepared at 40-80 wt % drug load. The drug and polymer blends were hot-melt extruded using a twin screw extruder through a 3 mm diameter die and pulled to a diameter of approximately 1.9-2.3 mm. The screws contained predominately conveying elements with a single 90° mixing section. The 1 st  zone where the drug-polymer blends were introduced was water-cooled and maintained at room temperature. The temperature for zones 2-4 was 100° C. for PLA and 75° C. for PCL. Extruded fibers with diameters between 1.9-2.3 mm were cut to the appropriate length to achieve the desired amount of drug per implant for in vivo studies. 
     All animal studies were conducted following protocols in accordance with the Institutional Animal Care and Use Committee (IACUC) at NIRC and Merck &amp; Co., Inc., Kenilworth, N.J., U.S.A. which adhere to the regulations outlined in the USDA Animal Welfare Act. For each implantation, a Wistar Han rat was anesthetized using isoflurane to effect prior to subcutaneous dose administrations. Using a trocar needle, the solid formulation (˜2 mm in diameter and of varying lengths based on the body weight of the individual animal to achieve the dose appropriate for each group) was placed in the scapular region. Four animals (2 males and 2 females) were used for each formulation. Animals were monitored until recovered. At indicated time points, samples of blood were obtained from anesthetized animals (using isoflurane) and processed to plasma for determination of 4′-ethynyl-2-fluoro-2′-deoxyadenosine levels. 
     
       
         
           
               
             
               
                 TABLE 4 
               
             
            
               
                   
               
               
                 4′-ethynyl-2-fluoro-2′-deoxyadenosine (EFdA) concentration in blood plasma from 40 wt %, 50 wt %,  
               
               
                 60 wt % and 80 wt % 4′-ethynyl-2-fluoro-2′-deoxyadenosine in poly(DL-lactide) (PLA) implants 
               
               
                 (n/a = not applicable, insufficient data for statistical analysis). 
               
            
           
           
               
               
               
               
               
            
               
                   
                 40 wt % EFdA + 
                 50 wt % EFdA + 
                 60 wt % EFdA + 
                 80 wt % EFdA + 
               
               
                   
                 60 wt % PLA 
                 50 wt % PLA 
                 40 wt % PLA 
                 20 wt % PLA 
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                   
                   
                 Std.  
                   
                 Std. 
                   
                 Std. 
                   
                 Std.  
               
               
                 Time 
                 Average 
                 Dev. 
                 Average 
                 Dev. 
                 Average 
                 Dev. 
                 Average 
                 Dev. 
               
               
                 (days) 
                 (nM) 
                 (nM) 
                 (nM) 
                 (nM) 
                 (nM) 
                 (nM) 
                 (nM) 
                 (nM) 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                 0.04 
                 323 
                 15 
                 614 
                 175 
                 1552 
                 694 
                 5055 
                 459 
               
               
                 0.08 
                 189 
                 9 
                 552 
                 171 
                 1458 
                 631 
                 5978 
                 988 
               
               
                 0.17 
                 73 
                 3 
                 372 
                 124 
                 1099 
                 381 
                 5243 
                 1028 
               
               
                 0.29 
                 46 
                 2 
                   
                   
                   
                   
                   
                   
               
               
                 1 
                 25 
                 5 
                 224 
                 80 
                 742 
                 253 
                 2095 
                 601 
               
               
                 2 
                 17 
                 2 
                 251 
                 90 
                 802 
                 191 
                 1750 
                 317 
               
               
                 3 
                 17 
                 2 
                 227 
                 80 
                 698 
                 167 
                 1478 
                 190 
               
               
                 4 
                 19 
                 3 
                 230 
                 77 
                 671 
                 133 
                 1403 
                 275 
               
               
                 7 
                 19 
                 2 
                 228 
                 81 
                 588 
                 113 
                 1398 
                 196 
               
               
                 10 
                 21 
                 1 
                 192 
                 58 
                 491 
                 87 
                 874 
                 207 
               
               
                 14 
                 22 
                 2 
                 188 
                 62 
                 445 
                 23 
                 353 
                 109 
               
               
                 17 
                 22 
                 2 
                 192 
                 59 
                 403 
                 55 
                 216 
                 54 
               
               
                 21 
                 19 
                 2 
                   
                   
                   
                   
                   
                   
               
               
                 22 
                   
                   
                 177 
                 49 
                 363 
                 60 
                 75 
                 48 
               
               
                 24 
                 19 
                 2 
                   
                   
                   
                   
                   
                   
               
               
                 25 
                   
                   
                 157 
                 35 
                 309 
                 63 
                 52 
                 15 
               
               
                 28 
                 20 
                 2 
                 155 
                 39 
                 229 
                 60 
                 56 
                 n/a 
               
               
                 31 
                 17 
                 2 
                 152 
                 42 
                 148 
                 91 
                 48 
                 n/a 
               
               
                 35 
                 16 
                 2 
                 150 
                 41 
                 92 
                 47 
                 19 
                 n/a 
               
               
                 38 
                   
                   
                 113 
                 33 
                 49 
                   
                   
                   
               
               
                 39 
                 18 
                 2 
                   
                   
                   
                   
                   
                   
               
               
                 42 
                   
                   
                 115 
                 25 
                   
                   
                   
                   
               
               
                 44 
                 17 
                 4 
                   
                   
                   
                   
                   
                   
               
               
                 45 
                   
                   
                 123 
                 16 
                   
                   
                   
                   
               
               
                 51 
                 15 
                 2 
                   
                   
                   
                   
                   
                   
               
               
                 53 
                   
                   
                 108 
                 16 
                   
                   
                   
                   
               
               
                 58 
                 13 
                 2 
                   
                   
                   
                   
                   
                   
               
               
                 60 
                   
                   
                 70 
                 9 
                   
                   
                   
                   
               
               
                 64 
                   
                   
                 62 
                 42 
                   
                   
                   
                   
               
               
                 65 
                 14 
                 1 
                   
                   
                   
                   
                   
                   
               
               
                 72 
                 13 
                 2 
                   
                   
                   
                   
                   
                   
               
               
                 74 
                   
                   
                 53 
                 44 
                   
                   
                   
                   
               
               
                 78 
                   
                   
                 48 
                 31 
                   
                   
                   
                   
               
               
                 79 
                 13 
                 2 
                   
                   
                   
                   
                   
                   
               
               
                 85 
                   
                   
                 67 
                 n/a 
                   
                   
                   
                   
               
               
                 86 
                 11 
                 1 
                   
                   
                   
                   
                   
                   
               
               
                 92 
                   
                   
                 56 
                 n/a 
                   
                   
                   
                   
               
               
                 93 
                 13 
                 3 
                   
                   
                   
                   
                   
                   
               
               
                 100 
                 11 
                 2 
                   
                   
                   
                   
                   
                   
               
               
                 106 
                   
                   
                 30 
                 n/a 
                   
                   
                   
                   
               
               
                 107 
                 10 
                 1 
                   
                   
                   
                   
                   
                   
               
               
                 113 
                   
                   
                 7 
                 n/a 
                   
                   
                   
                   
               
               
                 114 
                 9 
                 2 
                   
                   
                   
                   
                   
                   
               
               
                 121 
                 7 
                 2 
                   
                   
                   
                   
                   
                   
               
               
                 128 
                 7 
                 2 
                   
                   
                   
                   
                   
                   
               
               
                 135 
                 7 
                 2 
                   
                   
                   
                   
                   
                   
               
               
                 142 
                 4 
                 3 
                   
                   
                   
                   
                   
                   
               
               
                 156 
                 5 
                 2 
                   
                   
                   
                   
                   
                   
               
               
                 163 
                 4 
                 1 
                   
                   
                   
                   
                   
                   
               
               
                 170 
                 3 
                 1 
                   
                   
                   
                   
                   
                   
               
               
                 177 
                 4 
                 n/a 
                   
                   
                   
                   
                   
                   
               
               
                 184 
                 2 
                 1 
                   
                   
                   
                   
                   
                   
               
               
                 191 
                 3 
                 n/a 
                   
                   
                   
                   
                   
                   
               
               
                 212 
                 23 
                 19 
                   
                   
                   
                   
                   
                   
               
               
                 219 
                 83 
                 77 
                   
                   
                   
                   
                   
                   
               
               
                 232 
                 6 
                 n/a 
                   
                   
                   
                   
                   
                   
               
               
                 239 
                 4 
                 n/a 
                   
                   
                   
                   
                   
                   
               
               
                 246 
                 3 
                 n/a 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 5 
               
             
            
               
                   
               
               
                 4′-ethynyl-2-fluoro-2′-deoxyadenosine (EFdA) in vivo release rates and fraction aborbed from 40 wt %, 
               
               
                 50 wt %, 60 wt % and 80 wt % 4′-ethynyl-2-fluoro-2′-deoxyadenosine in poly(DL-actide) (PLA) implants  
               
               
                 (normalized to a 40 mm long and 2 mm diameter implant) 
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                   
                 Release 
                 Fraction 
                 Release 
                 Fraction 
                 Release 
                 Fraction 
                 Release 
                 Fraction 
               
               
                   
                 Rate at 
                 absorbed 
                 Rate at 
                 absorbed 
                 Rate at 
                 absorbed 
                 Rate at 
                 absorbed 
               
               
                   
                 day 3 
                 at day 3 
                 day 30 
                 at day 30 
                 day 60 
                 at day 60 
                 day 127 
                 at day 127 
               
               
                 Sample 
                 (mg/day) 
                 (%) 
                 (mg/day) 
                 (%) 
                 (mg/day) 
                 (%) 
                 (mg/day) 
                 (%) 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                 40 wt % EFdA + 
                 0.15 
                 1.3 
                 0.16 
                 8.6 
                 0.12 
                 15.0 
                 0.06 
                 25.0 
               
               
                 60 wt % PLA 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 50 wt % EFdA + 
                 0.92 
                 3.5 
                 0.62 
                 26.5 
                 0.28 
                 42.4 
                 n/a 
                 42.4 
               
               
                 50 wt % PLA 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 60 wt % EFdA + 
                 2.74 
                 9.4 
                 0.69 
                 53 
                 n/a 
                 53 
                 n/a 
                 53 
               
               
                 40 wt % PLA 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 80 wt % EFdA + 
                 6.12 
                 21.7 
                 0.21 
                 60.7 
                 n/a 
                 60.7 
                 n/a 
                 60.7 
               
               
                 20 wt % PLA 
                   
                   
                 (day 25) 
                   
                   
                   
                   
                   
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 6 
               
             
            
               
                   
               
               
                 4′-ethynyl-2-fluoro-2′-deoxyadenosine (EFdA) concentration  
               
               
                 in blood plasma from 40 wt % and 60 wt % 4′-ethynyl-2-fluoro-  
               
               
                 2′-deoxyadenosine in poly(caprolactone) (PCL) implants  
               
               
                 (n/a = not applicable, insufficient data for statistical analysis). 
               
            
           
           
               
               
               
            
               
                   
                 40 wt % EFdA + 
                 60 wt % EFdA + 
               
               
                   
                 60 wt % PCL 
                 40 wt % PCL 
               
            
           
           
               
               
               
               
               
            
               
                 Time 
                 Average 
                 Std. Dev. 
                 Average 
                 Std. Dev. 
               
               
                 (days) 
                 (nM) 
                 (nM) 
                 (nM) 
                 (nM) 
               
               
                   
               
            
           
           
               
               
               
               
               
            
               
                 0.04 
                 158 
                 57 
                 5353 
                 911 
               
               
                 0.08 
                 130 
                 30 
                 5635 
                 899 
               
               
                 0.17 
                 85 
                 26 
                 4230 
                 996 
               
               
                 0.29 
                 55 
                 14 
                   
                   
               
               
                 1 
                 33 
                 7 
                 931 
                 299 
               
               
                 2 
                 27 
                 4 
                 664 
                 273 
               
               
                 3 
                 21 
                 2 
                 455 
                 146 
               
               
                 4 
                 20 
                 3 
                 359 
                 90 
               
               
                 7 
                 14 
                 1 
                 205 
                 37 
               
               
                 10 
                 12 
                 2 
                 127 
                 21 
               
               
                 14 
                 11 
                 2 
                 81 
                 4 
               
               
                 17 
                 10 
                 2 
                 74 
                 5 
               
               
                 21 
                 10 
                 1 
                   
                   
               
               
                 22 
                   
                   
                 63 
                 8 
               
               
                 24 
                 9 
                 1 
                   
                   
               
               
                 25 
                   
                   
                 53 
                 6 
               
               
                 28 
                 9 
                 2 
                 49 
                 4 
               
               
                 31 
                 8 
                 1 
                 47 
                 4 
               
               
                 35 
                 8 
                 2 
                 45 
                 3 
               
               
                 38 
                   
                   
                 34 
                 3 
               
               
                 39 
                 6 
                 1 
                   
                   
               
               
                 42 
                   
                   
                 34 
                 6 
               
               
                 44 
                 6 
                 0 
                   
                   
               
               
                 45 
                   
                   
                 32 
                 2 
               
               
                 51 
                 5 
                 0 
                   
                   
               
               
                 53 
                   
                   
                 34 
                 2 
               
               
                 58 
                 4 
                 1 
                   
                   
               
               
                 60 
                   
                   
                 22 
                 3 
               
               
                 64 
                   
                   
                 26 
                 4 
               
               
                 65 
                 4 
                 0 
                   
                   
               
               
                 72 
                 4 
                 0 
                   
                   
               
               
                 74 
                   
                   
                 24 
                 3 
               
               
                 78 
                   
                   
                 19 
                 4 
               
               
                 79 
                 4 
                 1 
                   
                   
               
               
                 85 
                   
                   
                 17 
                 3 
               
               
                 86 
                 3 
                 0 
                   
                   
               
               
                 92 
                   
                   
                 18 
                 3 
               
               
                 93 
                 3 
                 1 
                   
                   
               
               
                 100 
                 3 
                 1 
                   
                   
               
               
                 106 
                   
                   
                 13 
                 2 
               
               
                 107 
                 2 
                 1 
                   
                   
               
               
                 113 
                   
                   
                 11 
                 2 
               
               
                 114 
                 2 
                 0 
                   
                   
               
               
                 120 
                   
                   
                 14 
                 3 
               
               
                 121 
                 2 
                 0 
                   
                   
               
               
                 127 
                   
                   
                 13 
                 1 
               
               
                 128 
                 2 
                 0 
                   
                   
               
               
                 134 
                   
                   
                 11 
                 1 
               
               
                 135 
                 2 
                 0 
                   
                   
               
               
                 141 
                   
                   
                 10 
                 1 
               
               
                 142 
                 2 
                 0 
                   
                   
               
               
                 148 
                   
                   
                 11 
                 2 
               
               
                 155 
                   
                   
                 9 
                 1 
               
               
                 156 
                 1 
                 1 
                   
                   
               
               
                 162 
                   
                   
                 8 
                 1 
               
               
                 163 
                 1 
                 1 
                   
                   
               
               
                 169 
                   
                   
                 9 
                 2 
               
               
                 170 
                 2 
                 1 
                   
                   
               
               
                 176 
                   
                   
                 9 
                 2 
               
               
                 177 
                 1 
                 0 
                   
                   
               
               
                 183 
                   
                   
                 7 
                 1 
               
               
                 184 
                 1 
                 0 
                   
                   
               
               
                 190 
                   
                   
                 7 
                 2 
               
               
                 191 
                 1 
                 1 
                   
                   
               
               
                 197 
                   
                   
                 7 
                 1 
               
               
                 204 
                   
                   
                 7 
                 2 
               
               
                 211 
                   
                   
                 7 
                 1 
               
               
                 212 
                 1 
                 0 
                   
                   
               
               
                 218 
                   
                   
                 7 
                 1 
               
               
                 219 
                 1 
                 0 
                   
                   
               
               
                 225 
                   
                   
                 7 
                 2 
               
               
                 232 
                 1 
                 0 
                 7 
                 1 
               
               
                 239 
                 1 
                 0 
                 5.8 
                 0.2 
               
               
                 246 
                 1 
                 1 
                 7 
                 1 
               
               
                 253 
                 1 
                 0 
                 7 
                 2 
               
               
                 260 
                   
                   
                 7 
                 1 
               
               
                 267 
                 1 
                 1 
                   
                   
               
               
                 269 
                   
                   
                 7 
                 2 
               
               
                 276 
                   
                   
                 6 
                 1 
               
               
                 274 
                 1 
                 0 
                   
                   
               
               
                 281 
                 1 
                 1 
                   
                   
               
               
                 283 
                   
                   
                 6 
                 1 
               
               
                 288 
                 1 
                 0 
                   
                   
               
               
                 290 
                   
                   
                 6 
                 1 
               
               
                 295 
                 2 
                 n/a 
                   
                   
               
               
                 297 
                   
                   
                 6 
                 1 
               
               
                 302 
                 1 
                 n/a 
                   
                   
               
               
                 304 
                   
                   
                 7 
                 1 
               
               
                 309 
                 1 
                 n/a 
                   
                   
               
               
                 311 
                   
                   
                 7 
                 1 
               
               
                 316 
                 1 
                 n/a 
                   
                   
               
               
                 318 
                   
                   
                 8 
                 2 
               
               
                 323 
                 1 
                 n/a 
                   
                   
               
               
                 325 
                   
                   
                 9 
                 1 
               
               
                 330 
                 1 
                 n/a 
                   
                   
               
               
                 332 
                   
                   
                 6 
                 1 
               
               
                 337 
                 1 
                 n/a 
                   
                   
               
               
                 339 
                   
                   
                 7 
                 1 
               
               
                 344 
                 1 
                 n/a 
                   
                   
               
               
                 346 
                   
                   
                 7 
                 1 
               
               
                 351 
                 1 
                 n/a 
                   
                   
               
               
                 353 
                   
                   
                 7 
                 1 
               
               
                 360 
                   
                   
                 7 
                 1 
               
               
                 367 
                   
                   
                 8 
                 1 
               
               
                 374 
                   
                   
                 7 
                 2 
               
               
                 381 
                   
                   
                 7 
                 1 
               
               
                 388 
                   
                   
                 7 
                 1 
               
               
                 395 
                   
                   
                 7 
                 1 
               
               
                 402 
                   
                   
                 7 
                 1 
               
               
                 409 
                   
                   
                 6 
                 1 
               
               
                 416 
                   
                   
                 5.2 
                 0.3 
               
               
                 423 
                   
                   
                 5 
                 1 
               
               
                 429 
                   
                   
                 5 
                 1 
               
               
                 437 
                   
                   
                 4 
                 1 
               
               
                 444 
                   
                   
                 5 
                 1 
               
               
                 451 
                   
                   
                 5 
                 1 
               
               
                 458 
                   
                   
                 5 
                 1 
               
               
                 465 
                   
                   
                 4 
                 1 
               
               
                 472 
                   
                   
                 4 
                 1 
               
               
                 479 
                   
                   
                 4 
                 1 
               
               
                 486 
                   
                   
                 3.3 
                 0.4 
               
               
                 493 
                   
                   
                 3.6 
                 0.4 
               
               
                 500 
                   
                   
                 4 
                 1 
               
               
                 507 
                   
                   
                 3.6 
                 0.3 
               
               
                 514 
                   
                   
                 3.2 
                 0.5 
               
               
                 521 
                   
                   
                 3 
                 1 
               
               
                 528 
                   
                   
                 4 
                 2 
               
               
                 535 
                   
                   
                 3 
                 1 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 7 
               
             
            
               
                   
               
               
                 4′-ethynyl-2-fluoro-2′-deoxyadenosine (EFdA) in vivo release rates and fraction absorbed from  
               
               
                 40% and 60% 4′-ethynyl-2-fluoro-2′-deoxyadenosine in poly(caprolactone) (PCL) implants 
               
               
                 (normalized to a 40 mm long and 2 mm diameter implant) 
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                   
                 Release 
                 Fraction 
                 Release 
                 Fraction 
                 Release 
                 Fraction 
                 Release 
                 Fraction 
               
               
                   
                 Rate at 
                 absorbed 
                 Rate at 
                 absorbed 
                 Rate at 
                 absorbed 
                 Rate at 
                 absorbed 
               
               
                   
                 day 3 
                 at day 3 
                 day 30 
                 at day 30 
                 day 60 
                 at day 60 
                 day 127 
                 at day 127 
               
               
                 Sample 
                 (mg/day) 
                 (%) 
                 (mg/day) 
                 (%) 
                 (mg/day) 
                 (%) 
                 (mg/day) 
                 (%) 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                 40 wt % 
                 0.58 
                 5.3 
                 0.23 
                 20.1 
                 0.12 
                 28.9 
                 0.05 
                 38.6 
               
               
                 EFdA + 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 60 wt % 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 PCL 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 60 wt % 
                 1.69 
                 15.6 
                 0.19 
                 26.9 
                 0.09 
                 30.8 
                 0.05 
                 35.1 
               
               
                 EFdA + 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 40 wt % 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 PCL 
               
               
                   
               
            
           
         
       
     
     Example 3 
     Preparation and In Vivo Studies of Implant Drug Delivery Systems Containing 45 Wt % 4′-Ethynyl-2-Fluoro-2′-Deoxyadenosine 
     Implantable devices were prepared using an extrusion process. The first step involved mixing the dry, micronized powders of the active compound (4′-ethynyl-2-fluoro-2′-deoxyadenosine) and the cryomilled PCL (Evonik RESOMER® Select 100 CL 7.5E) or PLA (Evonik RESOMER® Select 100 DL 8A) using a Turbula T2F mixer. Drug and polymer blends were prepared at 40-80 wt % drug load. The drug and polymer blends were hot-melt extruded using a twin screw extruder through a 3 mm diameter die and pulled to a diameter of approximately 1.9-2.3 mm. The screws contained predominately conveying elements with a single 90° mixing section. The 1 st  zone where the drug-polymer blends were introduced was water-cooled and maintained at room temperature. The temperature for zones 2-4 was 100° C. for PLA and 75° C. for PCL. Extruded fibers with diameters between 1.9-2.3 mm were cut to the appropriate length to achieve the desired amount of drug per implant for in vivo studies. 
     All animal studies were conducted following protocols in accordance with the Institutional Animal Care and Use Committee (IACUC) at NIRC and Merck, which adhere to the regulations outlined in the USDA Animal Welfare Act. For each implantation, a Rhesus monkey was sedated with Ketamine HCl (100 mg/mL) prior to subcutaneous implant administrations. Using an injector device, the implant was placed subcutaneously in the interscapular region. Either two or three animals were used per group (n=2 for 40 wt % EFdA+60 wt % PLA, n=3 for 50 wt % EFdA+50 wt % PLA, n=2 for 50 wt % EFdA+50 wt % PCL). Animals were monitored until recovered. At indicated time points, samples of blood were obtained and processed to plasma for determination of 4′-ethynyl-2-fluoro-2′-deoxyadenosine levels. 
     
       
         
           
               
             
               
                 TABLE 8 
               
             
            
               
                   
               
               
                 4′-ethynyl-2-fluoro-2′-deoxyadenosine (EFdA) concentration  
               
               
                 in blood plasma from 40 wt % 4′-ethynyl-2-fluoro-2′- 
               
               
                 deoxyadenosine in poly(DL-lactide) (PLA) implants and 50 wt %  
               
               
                 4′-ethynyl-2-fluoro-2′-deoxyadenosine in PLA implants 
               
            
           
           
               
               
               
            
               
                   
                 40 wt % EFdA + 
                 50 wt % EFdA + 
               
               
                   
                 60 wt % PLA 
                 50 wt % EVA 
               
            
           
           
               
               
               
               
               
            
               
                 Time  
                 Avg 
                 std. dev. 
                 Avg 
                 std. dev. 
               
               
                 (days) 
                 (nM) 
                 (nM) 
                 (nM) 
                 (nM) 
               
               
                   
               
            
           
           
               
               
               
               
               
            
               
                 0.0833 
                 98 
                 2 
                 67 
                 34 
               
               
                 0.1667 
                 69 
                 5 
                 55 
                 26 
               
               
                 0.25 
                 56 
                 5 
                 51 
                 21 
               
               
                 1 
                 53.6 
                 0.4 
                 62 
                 23 
               
               
                 2 
                 42 
                 4 
                 44 
                 15 
               
               
                 9 
                 26 
                 5 
                 37 
                 13 
               
               
                 16 
                 22 
                 6 
                 35 
                 8 
               
               
                 30 
                 16 
                 1 
                 25 
                 5 
               
               
                 44 
                 7 
                 4 
                 17 
                 4 
               
               
                 58 
                 5 
                 4 
                 11 
                 3 
               
               
                 72 
                 5 
                 3 
                 7 
                 2 
               
               
                 86 
                 4 
                 3 
                 4 
                 1 
               
               
                 100 
                 4 
                 3 
                 1.2 
                 0.2 
               
               
                 114 
                 3 
                 3 
                 n/a 
                 n/a 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 9 
               
             
            
               
                   
               
               
                 4′-ethynyl-2-fluoro-2′-deoxyadenosine (EFdA) in vivo  
               
               
                 release rates from 40 wt % and 50 wt % 4′-ethynyl-2-fluoro- 
               
               
                 2′-deoxyadenosine in poly(DL-lactide) (PLA) implants 
               
            
           
           
               
               
               
               
            
               
                   
                 Release rate  
                 Release rate  
                 Release rate  
               
               
                   
                 at day30  
                 at day 60  
                 at day 114  
               
               
                 Sample 
                 (mg/day) 
                 (mg/day) 
                 (mg/day) 
               
               
                   
               
               
                 40 wt % EFdA + 
                 0.40-0.74 
                 0.14-0.26  
                 0.07-0.18 
               
               
                 60 wt % PLA 
                   
                   
                   
               
               
                 50 wt % EFdA + 
                 0.69-1.29 
                 0.30-0.55 
                 n/a 
               
               
                 50 wt % PLA 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 10 
               
             
            
               
                   
               
               
                 4′-ethynyl-2-fluoro-2′-deoxyadenosine (EFdA) concentration 
               
               
                 in blood plasma from 50 wt % 4′-ethynyl-2-fluoro-2′- 
               
               
                 deoxyadenosine in poly(caprolactone) (PCL) implants 
               
            
           
           
               
               
            
               
                   
                 50 wt % EFdA + 
               
               
                   
                 50 wt % PCL 
               
            
           
           
               
               
               
            
               
                 Time 
                 Avg 
                 std. dev. 
               
               
                 (days) 
                 (nM) 
                 (nM) 
               
               
                   
               
            
           
           
               
               
               
            
               
                 0.0833 
                 37 
                 2 
               
               
                 0.1667 
                 33 
                 7 
               
               
                 0.25 
                 27 
                 2 
               
               
                 1 
                 37 
                 11 
               
               
                 2 
                 21 
                 4 
               
               
                 9 
                 12 
                 3 
               
               
                 16 
                 9 
                 1 
               
               
                 30 
                 6 
                 1 
               
               
                 44 
                 5 
                 2 
               
               
                 58 
                 5 
                 1 
               
               
                 72 
                 4 
                 1 
               
               
                 86 
                 3.1 
                 0.3 
               
               
                 100 
                 2.6 
                 0.1 
               
               
                 114 
                 2.5 
                 0.1 
               
               
                 128 
                 2.2 
                 0.2 
               
               
                 142 
                 1.7 
                 0.0 
               
               
                 156 
                 1.5 
                 0.2 
               
               
                 170 
                 1.4 
                 0.2 
               
               
                 184 
                 1.4 
                 0.2 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 11 
               
             
            
               
                   
               
               
                 4′-ethynyl-2-fluoro-2′-deoxyadenosine (EFdA) in vivo  
               
               
                 release rates from 50 wt % 4′-ethynyl-2-fluoro-2′- 
               
               
                 deoxyadenosine in poly(caprolactone) (PCL) implants 
               
            
           
           
               
               
               
               
            
               
                   
                 Release rate  
                 Release rate  
                 Release rate  
               
               
                   
                 at day 30  
                 at day 60  
                 at day 114  
               
               
                 Sample 
                 (mg/day) 
                 (mg/day) 
                 (mg/day) 
               
               
                   
               
               
                 50 wt % EFdA + 
                 0.22-0.47 
                 0.18-0.38 
                 0.09-0.19 
               
               
                 50 wt % PCL