Long-acting colloidal pharmaceutical composition of an integrase chain transfer inhibitor and related method

Abstract

A composition comprising a stable colloid of an HIV integrase inhibitor for the preparation of an injectable, long-acting pharmaceutical product for treating and preventing HIV infection.

Description

Technical Field

[0001] (Cross - Reference to Related Applications) This application claims the benefit of U.S. Patent Application No. 63 / 498461, filed on April 26, 2023, which is hereby expressly incorporated by reference in its entirety.

[0002] (Statement of Government Licensing Rights) This invention was made with government support under Contract No. R61 / 33 AI149665, awarded by the National Institutes of Health. The government has certain rights in this invention.

Background Art

[0003] Successful advances in the development of many effective drug substances have made it possible to introduce single-dose or multi-dose oral drug products to maximize inhibition of HIV viral replication. However, all available oral drug products require at least daily or even more frequent administration in HIV+ patients to suppress the virus. People who have survived HIV can expect to live virus-free for a long period of time, provided they take tablets daily for an extended period. Daily HIV tablets are called highly active antiretroviral therapy (HAART) and consist of a combination of antiretroviral drugs, often referred to as cART (combinations anti-retroviral therapeutics). cART is intended to block HIV replication using drug substances that are designed to maximize inhibition of viral replication by inhibiting multiple replication checkpoints. HAART or cART typically consists of two to three drug substances intended to inhibit viral reverse transcription (RT), proteases, and / or integrases (often called integrase strand transfer inhibitors or INSTIs). Thanks to the global implementation of HAART and global efforts to identify, treat, and achieve effective viral suppression with cART, people living with HIV can now live into old age. However, long-term daily oral pills can cause pill fatigue. Stopping cART can lead to viral rebound, disease progression to AIDS, and premature death. Interruption of treatment and non-adherence to prescribed daily doses increase the likelihood of drug-resistant viruses in patients who are also at risk of viral rebound and progression to AIDS. While the current one-pill-a-day regimen has been proven to save lives, the WHO reported that in 2021, approximately 1.5 million people were newly infected with HIV, and more than 500,000 (approximately 650,000) died from HIV.

[0004] To overcome tablet fatigue, avoid preventable deaths, and address the aforementioned challenges, HIV drugs with long-acting (LA) pharmacokinetic properties that increase the dosing interval are being developed to improve treatment adherence and reduce tablet burden. A typical approach involves creating long-acting HIV drug products, which include selecting water-insoluble derivatives that exhibit longer terminal phase half-lives in the blood to produce injectable dosage forms that sustain drug levels for weeks to months. For example, the long-acting cabotegravir (LA-CAB, INSTI) product incorporated into CABENUVA, recently approved by the FDA [consisting of two drug products, LA cabotegravir injection and LA rilpivirine (LA-RPV, RT inhibitor) injection, administered intramuscularly to two separate sites]. Both LA-CAB and LA-RPV are manufactured using excipients such as PEG400 and poloxamer (polymer compounds) to form small drug crystals suitable for injection. These products retain their natural crystalline state. Nanocrystalline or nanocrystals are granulated to improve the solubility of water-insoluble drugs. Consequently, nanodrug crystal platforms are typically used to enhance drug dissolution rates by reducing the size of large, naturally occurring particulate drug crystals, thereby increasing oral absorption. However, nanocrystals are metastable and tend to invert into larger, more stable structures of the polymorphic crystalline form of the drug. Metastable nanocrystalline drugs are known to be unstable and have increased water solubility. Therefore, there is an urgent need for the formation of stable drug particles that achieve sustained plasma drug levels over extended periods, or for long-acting drug products based on novel and non-trivial compositions and methods. The goal is to provide pharmaceutically acceptable injectable products that are stable, scalable, and easily resuspendable, and further, to achieve long-lasting plasma drug concentrations to overcome the requirement of daily oral administration.Even with long-acting or sustained-release drug formulations packaged in oral dosage forms, the drug is eliminated from the gastrointestinal tract (intestines) in humans within approximately 24 hours. Therefore, daily tablet administration is still necessary to achieve drug levels effective in suppressing viral replication.

[0005] In some embodiments, methods and techniques for producing long-acting drug products are described. Some techniques include (1) conjugating drug molecules into an erosive biopolymer that releases drug molecules as the drug molecules are separated from the polymer over time; (2) encapsulating biodegradable polymer (e.g., PLGA or PLA) particles or drug molecules in a sustained-release hydrogel; (3) encapsulating in lipid vesicles, silica, clay and other carriers; and (4) grinding large drug crystals in an insoluble suspension containing polymer excipients to form small nanocrystalline drug products that dissolve slowly upon injection.

[0006] In one embodiment, current methods for producing long-acting and sustained release of drugs require cumbersome and time-consuming chemical modifications of the drug substance. Some encapsulation processes may involve mechanical milling processes that require the removal of non-encapsulated material or may result in contamination, heat-induced decomposition, increased costs, and increased production risks.

[0007] According to the WHO, 38.4 million people were living with HIV in 2021, a number that has continued to increase by approximately 1 million from the previous year. Therefore, there is an urgent need to develop simple, scalable, long-acting HIV products that are easy to manufacture, distribute, and use by people with HIV and those at risk of exposure.

[0008] The disclosures described herein provide novel compositions and simple methods that enable the production of stable and suitable colloidal drug products for creating injectable pharmaceutical suspension products to achieve the presence of long-acting drugs in the body. [Overview of the project]

[0009] In one embodiment, the present disclosure provides colloidal integrase chain transfer inhibitor particles comprising an integrase chain transfer inhibitor and an amino acid. Representative integrase chain transfer inhibitors useful in particles include dolutegravir, bictegravir, cabotegravir, raltegravir, and elvitegravir. Representative amino acids useful in particles include glutamine and tryptophan.

[0010] In another embodiment, the present disclosure provides an injectable pharmaceutical composition comprising a suspension of colloidal integrase chain transfer inhibitor particles and an aqueous carrier as described herein.

[0011] In a further embodiment, the Disclosure provides a method for treating or preventing HIV in a subject, comprising the step of administering to a subject in need of such treatment or prevention a therapeutically effective amount of integrase chain transfer inhibitor particles or an injectable pharmaceutical composition as described herein.

[0012] In another aspect, the Disclosure provides a method for treating a disease or condition that is preventable or treatable by administering an integrase chain transfer inhibitor, the method comprising the step of administering a therapeutically effective amount of an integrase chain transfer inhibitor particle or an injectable pharmaceutical composition described herein to a subject in need thereof. [Brief explanation of the drawing]

[0013] The above-described aspects of the present invention and its many associated advantages will become more readily understood as they are better understood in conjunction with the accompanying drawings and by referring to the following detailed description. [Figure 1] This is a schematic diagram illustrating the binding of an integrase chain transfer inhibitor (INSTI) to the integrase catalytic site of the HIV integrase enzyme (e.g., E152, D64, and D116) via cross-linked Mg2+ ions, using three representative coplanar oxygen atoms in the inhibitor. [Figure 2] This diagram compares the chemical structures of representative integrase chain transfer inhibitors, each having three sets of oxygen atoms (circled) on the same plane. The phenyl halide ring substituents are also circled. [Figure 3A] This is an image of a typical colloidal integrase chain transfer inhibitor particle containing dolutegravir (DTG) and glutamine (Gln). This product is called the DTG-CS product and has a final molar ratio of glutamine (Gln) to dolutegravir (DTG) of 9:2. The particle shape is spherical and has a diameter of approximately 354 nm. The particle size can be adjusted to produce a range of sizes based on the Gln to DTG ratio and the Gln addition ratio. [Figure 3B] This is an image of dolutegravir nanocrystals with a diameter of approximately 10 μm. [Figure 3C] This compares the small-angle X-ray diffraction analysis of DTG-CS particles (DTG-CS, Gln:DTG=9.2) (upper curve), a mixture of DTG and Gln (Gln:DTG=9.2) (middle curve), and the difference (lower curve). [Figure 4A] This image compares representative colloidal integrase chain transfer inhibitor particles: dolutegravir (DTG) and glutamine (Gln) (Figure 4A); and dolutegravir (DTG) and tryptophan (Trp) (Figure 4B). [Figure 4B] This image compares representative colloidal integrase chain transfer inhibitor particles: dolutegravir (DTG) and glutamine (Gln) (Figure 4A); and dolutegravir (DTG) and tryptophan (Trp) (Figure 4B). [Figure 5]This report describes the time-course (weekly) pharmacokinetics of dolutegravir concentration (ng / mL) in three non-human primates (NHPs) after a single subcutaneous injection of 5 mg / kg of DTG-CS particles (DTG-CS, Gln:DTG=9.2 m / m). A single injection of DTG in the CS formulation resulted in a long-term effect on the plasma drug concentration time course of dolutegravir. The presented data were obtained from two non-human primates, Southern pig-tailed macaques (M. Nemestrina). No significant adverse effects were observed in NHPs over 15 weeks. For comparison, the plasma time course of soluble DTG at 5 mg / kg after a single subcutaneous administration in NHPs is also presented. [Figure 6] These are images of typical colloidal integrase chain transfer inhibitor particles: bictegravir (BIC) and glutamine (Gln). BIC-CS particles (BIC-CS, Gln:BIC = 9:2 m / m) have a diameter of approximately 2 to 300 nm. [Figure 7] This study compares DTG-CS particle size (nm) as a function of glutamine concentration (%wt / v). Changing the DTG:Gln ratio reveals a direct relationship between Gln concentration (%wt / v) and particle size. Increasing the Gln concentration resulted in an increase in DTG-Gln CS particle size across the tested Gln% range. [Figure 8] This study demonstrates the formation of bictegravir particles (BIC-CS) as a function of glutamine concentration. A fixed concentration of 2.23 mM bictegravir in solution was to which the glutamine concentration was increased. An immediate increase in bictegravir particle formation was observed and completed within 1-2 minutes. The particles formed in the apparent turbid suspension were quantified using a spectrophotometer set to measure absorbance at 700 nm and expressed as A700. Higher absorbance indicates a greater degree of bictegravir particle formation in the suspension. This data is expressed as the molar ratio of glutamine to bictegravir. [Modes for carrying out the invention]

[0014] The present disclosure provides a composition comprising a stable colloid of an HIV integrase inhibitor for making an injectable long - acting pharmaceutical product for treating and preventing HIV infection. When injected into a subject, the composition achieves a long - term plasma profile of the HIV integrase inhibitor for over 15 weeks.

[0015] In one aspect, the present disclosure provides colloidal integrase strand transfer inhibitor particles comprising an integrase strand transfer inhibitor and an amino acid.

[0016] As used herein, the term "colloidal integrase strand transfer inhibitor particles" refers to particles comprising a mixture of a single species of small particles (i.e., an INSTI inhibitor) that is substantially uniformly distributed throughout another substance (i.e., an amino acid). Colloidal particles are generally larger than the size of a drug substance completely dissolved in solution and smaller than the size in a drug suspension.

[0017] As used herein, the term "integrase strand transfer inhibitor" (INSTI) refers to a compound that inhibits the HIV integrase enzyme by binding to the integrase catalytic site (e.g., E152, D64, and / or D116) by three sets of oxygen atoms on the same plane in the inhibitor via a bridging Mg2 + ion, as schematically shown in FIG. 1. The integrase strand transfer inhibitors described herein have three sets of oxygen atoms on the same plane as shown in FIGS. 1 and 2. FIG. 2 illustrates the chemical structure of a representative integrase strand transfer inhibitor having three sets of oxygen atoms (each surrounded by a circle) on the same plane. The representative three sets of oxygen atoms on the same plane include the following moieties of formulas (I), (IA), (IB), and (II):

[0018]

Chemical formula

[0019] Representative integrase chain transfer inhibitors described herein include dolutegravir, bictegravir, cabotegravir, raltegravir, and elvitegravir. Dolutegravir, bictegravir, and cabotegravir contain portions of formulas (I) and (IA), raltegravir contains portion of formula (IB), and elvitegravir contains portion of formula (II).

[0020] The colloidal integrase chain transfer inhibitor particles described herein contain amino acids in addition to the integrase chain transfer inhibitor. The amino acids stabilize the colloidal particles. As described herein and without being bound by theory, the colloidal particles described herein appear to be held together not by ionic interactions between the integrase chain transfer inhibitor (e.g., DTG) and the amino acids (e.g., glutamine and tryptophan), but by hydrogen bonding (H-bonding) interactions, and maintained within the colloidal particle structure by van der Waals forces. The H-bonding interaction between the amino acids and the integrase chain transfer inhibitor is thought to be H-bonds between the three sets of oxygen atoms on the same plane as the amino acids and the integrase chain transfer inhibitor (see Figure 2, and parts of formulas (I), (IA), (IB), and (II) containing three sets of oxygen atoms on the same plane). The halogenated phenyl ring of INSTI may also potentially interact with the phenyl ring of tryptophan via π-π electron interactions.

[0021] The colloidal integrase chain transfer inhibitor particles described herein are effective for long-term delivery of integrase chain transfer inhibitors, having advantageous pharmacokinetic properties and profiles compared to the individual integrase chain transfer inhibitors alone, and being released over time from the particles. Due to their stable colloidal form, the integrase chain transfer inhibitor particles described herein are long-acting particles for delivering the integrase chain transfer inhibitors they contain.

[0022] In certain embodiments, the integrase chain transfer inhibitor is selected from the group consisting of dolutegravir, bictegravir, cabotegravir, raltegravir, and elvitegravir. In other embodiments, the integrase chain transfer inhibitor is selected from the group consisting of dolutegravir, bictegravir, and cabotegravir. In further embodiments, the integrase chain transfer inhibitor is selected from the group consisting of raltegravir and elvitegravir. In one embodiment, the integrase chain transfer inhibitor is dolutegravir.

[0023] As stated above, the colloidal integrase chain transfer inhibitor particles described herein contain an amino acid in addition to the integrase chain transfer inhibitor. In certain embodiments, the amino acid is glutamine. In other embodiments, the amino acid is tryptophan. Suitable amino acids include L-isomers, D-isomers, and racemic amino acids.

[0024] In certain embodiments of the colloidal particles, the integrase chain transfer inhibitor is dolutegravir (DTG), and the stabilizing amino acid is glutamine (Gln). In some of these embodiments, the molar ratio of dolutegravir to glutamine (DTG:Gln) is 1:0.5–12 m / m. In other embodiments, the molar ratio of dolutegravir to glutamine (DTG:Gln) is 1:4.9 m / m.

[0025] In other embodiments of the colloidal particles, the integrase chain transfer inhibitor is dolutegravir (DTG), and the stabilizing amino acid is tryptophan (Trp). In some of these embodiments, the molar ratio of dolutegravir to tryptophan (DTG:Trp) is 1:0.5–12 m / m. In other embodiments, the molar ratio of dolutegravir to glutamine (DTG:Trp) is 1:4.9 m / m.

[0026] In further embodiments of the colloidal particles, the integrase chain transfer inhibitor is bictegravir (BIC), and the stabilizing amino acid is glutamine (Gln). In some of these embodiments, the molar ratio of bictegravir to glutamine (BIC:Gln) is 1:0.5–12 m / m. In other embodiments, the molar ratio of bictegravir to glutamine (BIC:Gln) is 1:5 m / m.

[0027] In other embodiments of the colloidal particles, the integrase chain transfer inhibitor is bictegravir (BIC), and the stabilizing amino acid is tryptophan (Trp). In some of these embodiments, the molar ratio of bictegravir to tryptophan (DTG:Trp) is 1:0.5–12 m / m.

[0028] The preparation of representative integrase chain transfer inhibitor particles (DTG:Gln) and their corresponding injectable compositions is described in Example 1. Other representative integrase chain transfer inhibitor particles (e.g., DTG:Trp and BIC:Gln) and their corresponding injectable compositions were prepared according to the procedures described in Example 1. Integrase chain transfer inhibitor particles containing cabotegravir, raltegravir, and elvitegravir, and their corresponding injectable compositions, are prepared according to the procedures described in Example 1.

[0029] In another embodiment, the present disclosure provides an injectable pharmaceutical composition comprising a suspension of colloidal integrase chain transfer inhibitor particles described herein in an aqueous solvent.

[0030] In certain embodiments, the injectable pharmaceutical composition (or formulation) contains about 1 to about 2 mg of an integrase chain transfer inhibitor per 1 mL.

[0031] In a particular embodiment, the injectable pharmaceutical composition (or formulation) contains about 10 to about 15 mg of amino acids per 1 mL.

[0032] In certain embodiments, the injectable pharmaceutical composition (or formulation) further comprises biocompatible pharmaceutical excipients. Suitable excipients include histidine, parabens, sucrose, lactose, Tween20, and Tween80.

[0033] In certain embodiments, the injectable pharmaceutical composition (or formulation) further comprises a pegylation modifier. Preferred pegylation modifiers include lipid-conjugated PEG having various MWs (range: 500-5000) of PEG and acylated PEG having various MWs (range: 500-5000) of PEG.

[0034] In a further embodiment, the disclosure provides a method for using colloidal particles and injectable pharmaceutical compositions.

[0035] In one embodiment, the present disclosure provides a method for treating or preventing HIV in a subject. In a particular embodiment, the method includes the step of administering a therapeutically effective amount of integrase chain transfer inhibitor particles or an injectable composition described herein to a subject in need.

[0036] It is recognized that certain integrase chain transfer inhibitors have been approved for both HIV prevention and HIV treatment. Some have been approved for HIV treatment only.

[0037] In another embodiment, the Disclosure provides a method for treating or preventing a disease or condition treatable by administering an integrase chain transfer inhibitor to a subject. In a particular embodiment, the method includes the step of administering a therapeutically effective amount of an integrase chain transfer inhibitor particle or an injectable composition described herein to a subject in need. Other conditions or diseases known to be treatable by integrase chain transfer inhibitors include certain leukemias and cancers associated with retroviral-induced oncogene activation.

[0038] In the above method, the integrase chain transfer inhibitor particles or injectable composition are administered by injection, such as intravenous, subcutaneous, or intramuscular injection. In certain embodiments, 0.5 to 6 mL of the injectable composition (formulation) described herein is administered by 1 to 4 subcutaneous or intramuscular injections. Administration may be at any interval between 7 and 365 days.

[0039] Those who may benefit from the above method include individuals at risk of HIV exposure, individuals living with HIV, individuals recently exposed to HIV, and individuals living with HIV who are unable to swallow tablets.

[0040] The following describes the preparation, evaluation, and use of representative colloidal integrase chain transfer inhibitor particles and their injectable compositions (formulations).

[0041] In one embodiment, this disclosure provides a long-acting HIV therapeutic product comprising an HIV integrase (INSTI) inhibitor (e.g., dolutegravir or DTG, current HIV drug substances). The therapeutic product described herein is an injectable colloidal pharmaceutical product (e.g., a colloidal DTG product). The colloidal pharmaceutical product comprises a colloid (e.g., a DTG colloid), which is different from current drug delivery technologies, including encapsulation in liposomes, nanocrystals, polymer conjugates or encapsulation, clay, carbon, silicon, and other structures or substances used to produce long-acting drug products. The DTG colloid disclosed herein is stable in suspension and is suitable for use as an injectable pharmaceutical product exhibiting long-acting pharmacokinetics in mammals. Dolutegravir is the current first-line HIV drug that effectively inhibits the integration of the HIV virus sequence into host human genomic DNA to produce HIV offspring.

[0042] The DTG colloid disclosed herein contains amino acids that stabilize the colloid. To determine whether glutamine (Gln), one of the essential amino acids of the protein building blocks in animals and humans, is effective in stabilizing the colloid, dolutegravir sodium (DTG) was dissolved in water at 20 g / L at 25°C. Glutamine (Gln) is also dissolved in water at 15 g / L at 25°C. In a glass container, 8 parts of DTG [20 g / L] were mixed with 2 parts of Gln (15 g / L) at 25°C. Immediately after mixing, the mixture formed a colloid exhibiting a cloudy appearance. These spherical particles were detectable and observable under a high-resolution polarizing microscope (see Figures 3A and 4A). The particles had a diameter of 354 nm. These particles were stable at room temperature or stored at 4°C and were not destabilized upon dilution (after the formation of DTG-CS). The resulting DTG-Gln particles are colloidal and have been confirmed to be a stable product; therefore, in this specification, they are also referred to as DTG-CS.

[0043] Further dose-response studies using various concentrations of Gln in a mixture of fixed-concentration DTG-sodium in an acidic solution (pH 5-6) confirmed that the above composition and mixing method yielded optimal results (confirming that all DTG in the solution was found in DTG-CS, and that no excess free DTG remained in the solution containing DTG-CS). The tested Gln:DTG range was a molar ratio of 0-10. Excess DTG or Gln results in the residue of either drug in the solution, requiring the removal of soluble DTG or Gln as a by-product. Once the molar ratio of DTG:Gln forming the DTG-CS product is defined, the removal of free drugs is not required, thus realizing a unique advantage in the production of DTG-CS products. Therefore, the time, cost, and resources required in the production of DTG-CS as a long-acting drug product are significantly reduced.

[0044] Considering the molecular weights of each component in an 8:2v / v mixture (20g / L DTG:15g / L Gln), the molar ratio of DTG to Gln is 2:9 (m / m). Therefore, unless otherwise specified, DTG-Gln products, or DTG-CS consisting of 2:9 m / m DTG to Gln, are used as representative examples in the descriptions.

[0045] X-ray diffraction techniques were used to analyze the physical properties of DTG-CS particles. These techniques allow for the exploration of new structural and organizational distinctions in the molecular arrangements of DTG and Gln. X-ray diffraction enables the elucidation of structural distinctions between natural DTG crystals, mixtures of DTG and Gln, or either DTG or Gln alone. As shown in Figure 3C, it was determined that the mixture of DTG and Gln (center curve) is distinct from DTG-CS (top curve).

[0046] However, when comparing the X-ray diffraction results for the mixture (Figure 3C, middle curve) with those for colloidal form of DTG-Gln (Figure 3C, upper curve), it was surprising to find that the DTG-Gln mixture powder exhibited an X-ray diffraction pattern different from that of the natural profile, which is not present in the mixture. Based on subtraction analysis versus intensity reduction analysis at 2θ angles (Figure 3C, lower curve), significant increases in intensity are observed at 2θ angles of 13.4, 16.3, 22.4, 25.9, 34.4, and 46.7, and significant decreases at 20.32, 24.6, and 29.6. Therefore, DTG-CS is physically and crystallographically distinct from DTG drug crystals and mixtures of DTG-Gln. DTG-CS is currently a novel structure that was previously unknown.

[0047] The stability of DTG-CS products prepared with either glutamine (Gln) or tryptophan (Trp) in the same ratio (2:9 (m / m)) was evaluated. These DTG-CS products were stable for long-term storage. These DTG-CS products were not destabilized even when loaded with ionic amino acids such as lysine or arginine in excess of 10 to 100 times the amount of glutamine or tryptophan in solution. Furthermore, the DTG-CS products were not destabilized by the addition of positively charged lysine (in ionic form) or negatively charged glutamic acid (in ionic form). Therefore, these data suggest that the interaction between DTG and Trp or Gln is not of an ionic nature.

[0048] To investigate the presence of H bonds, which are proposed as key attributes of INSTI, and their interaction with nonionic amino acids such as glutamine or tryptophan, DTG-CS products were tested with urea, a well-understood H-bond disruptor commonly known as a chaotropic agent. Urea is known to protein scientists as a chaotropic agent that denatures the structure of proteins composed of specific amino acid polymer sequences folded and held together by hydrogen bonds and van der Waals forces, resulting in a three-dimensional structure for specific functions. Urea can disrupt these bonds, causing proteins to unfold or denature, thereby disrupting H-bond (non-covalent) interactions and van der Waals interactions between amino acids. Surprisingly, the DTG-CS products became unstable when urea concentrations were increased and added to them. The DTG-CS products were destabilized, and only at very high urea concentrations (15-20% w / v) did the DTG-CS products (opaque in appearance) revert to soluble DTG (soluble and transparent in appearance). There were some differences in sensitivity to urea between DTG-CS products prepared using Gln and Trp, and some became unstable, but the DTG-CS products were resistant to a maximum of 10-15 w / v% urea chaotropic agent loading. It appears that the DTG-CS products are held together not by ionic bonding, but probably by H bonds between DTG and stabilizers such as the amino acids Trp and Gln, and maintained within the DTG-CS structure by van der Waals forces.

[0049] In the preparation of pharmaceutically acceptable particulate products, excipients or injectable colloidal drug products intended to reduce self-aggregation may interfere with the product's ability to form or resuspend before use. After testing a range of common excipients and suitable surfactants, three pharmaceutical excipients were selected: dextrose, sucrose, and a less common excipient, polyethylene glycol polymer (MW=2000; or mPEG). 2000 )(Generally, mPEG 2000-DSPE (conjugated to disteroylphosphatidyethanolamine (DSPE)) was determined not to interfere with DTG-CS formation or the suspension of the resulting particles.

[0050] To determine the ability to form a DTG-CS suspension, a DTG-CS suspension was prepared using a gln stabilizer with 2 mg / ml DTG. In the preparation of the injectable DTG-CS formulation, 0.1% sucrose, 0.1% dextrose, or 0.014% pegylated lipid (mPEG) was used. 2000 The formation of DTG-CS was determined by adding mPEG2000-DSPE. After recovering the DTG-CS particles by centrifugation at 500g for 10 minutes, they were resuspended in water at 1 / 10 of their original volume. The results showed that the listed pharmaceutical excipients did not hinder the formation of DTG-CS, but in the case of injectable DTG-CS pharmaceutical preparations, the presence of 0.014 w / v% mPEG2000-DSPE in 2 mg / mL of DTG-CS improved the resuspendability of DTG-CS.

[0051] Colloidal bictegravir particles (BIC-CS) were also prepared. Figure 6 shows an image of BIC-CS particles prepared using glutamine as the amino acid. As shown in Figure 6, BIC-CS particles (BIC-CS, Gln:BIC=9:2) have a diameter of approximately 2 to 300 nm.

[0052] Figure 8 shows the formation of bictegravir particles (BIC-CS) as a function of glutamine concentration. Referring to Figure 8, glutamine was added to a fixed concentration of 2.23 mM bictegravir in solution, with increasing concentrations. An immediate increase in bictegravir particle formation was evident and completed within 1-2 minutes. The particles formed in the apparent turbid suspension were quantified using a spectrophotometer set to measure absorbance at 700 nm and expressed as A700. Higher absorbance indicates a greater degree of bictegravir particle formation in the suspension. The data are expressed as the molar ratio of glutamine to bictegravir.

[0053] As stated above, this disclosure provides an injectable, long-acting HIV therapeutic product comprising an HIV integrase (INSTI) inhibitor (e.g., dolutegravir or DTG, which are currently used as HIV drug substances). The DTG colloid disclosed herein is stable in suspension and is suitable for use as an injectable pharmaceutical product exhibiting long-acting pharmacokinetics in mammals. Figure 5 shows the pharmacokinetics of dolutegravir concentration (ng / mL) over time (weekly) in three non-human primates (NHPs) after a single subcutaneous injection of 5 mg / kg of DTG-CS particles (DTG-CS, Gln:DTG=9.2 m / m). A single injection of DTG in the CS formulation resulted in a long-acting plasma drug concentration time course of dolutegravir. The presented data were obtained in two non-human primates, Southern pig-tailed macaques. No significant adverse effects were observed in NHPs over a 15-week period. For comparison, the plasma time course of soluble DTG at 5 mg / kg after a single subcutaneous administration in NHPs is presented. Free drug (DTG) was prepared as a 1.6 mg / mL solution and administered to NHPs via the subcutaneous route at an equivalent dose of 5 mg / kg. Plasma drug concentrations were measured by LC-MS / MS assay (the same assay was used for NHP plasma samples collected from NHPs that received a subcutaneous injection of DTG equivalent to 5 mg / kg in the DTG-CS dosage form). The data shown are the average values ​​of the two NHPs. Plasma drug levels were below the detection limit of the LC-MS / MS assay until day 2.

[0054] In this specification, the term "approximately" refers to ±5% of the specified value.

[0055] The following embodiments are illustrative but not limiting to the present disclosure. [Examples]

[0056] Example 1 Preparation of typical colloidal integrase chain transfer inhibitor particles In this example, the preparation of representative colloidal integrase chain transfer inhibitor particles is described, including dolutegravir:glutamine (DTG-Gln CS) particles and their injectable formulations.

[0057] In this embodiment, the terms "DTG-Gln CS" and "DTG-CS" are used interchangeably.

[0058] An initial DTG-Gln particle suspension of DTG-CS containing approximately 1.4 mL of DTG in the DTG-CS dosage form was prepared using the following substances, compositions, and methods.

[0059] reagent Dolutegravir sodium salt (DTG) CAS#1051375-19-9. MW: 419.38 (Cipla). Glutamine (Gln): CAS#56-85-9, MW: 146.1 (Sigma). mPEG 2000 -DSPE sodium salt: CAS #247925-28-6, MW: 2805.5 (Lipoid) Sterile water for injection

[0060] Preparation of preservation solution A 2 mg / mL storage solution of DTG was prepared by adding 2 mg of DTG to 1 mL of water.

[0061] A 21 mg / mL glutamine storage solution was prepared by adding 21 mg of glutamine powder to 1 mL of water.

[0062] mPEG 2000 -DSPE 7 mg / mL storage solution is made by adding 7 mg of mPEG to 1 mL of water. 2000 - Prepared by adding DSPE.

[0063] Regarding the sterile injectable products, all three items underwent sterilization filtration, and the formulation process was carried out in a sterile hood.

[0064] Formulation of DTG-CS in suspension at 25°C To 600 mL of DTG (2 mg / mL) solution, 120 mL of glutamine (21 mg / mL) was added and mixed using a stirrer at 60 rpm.

[0065] Next, within 1-2 minutes, continue mixing while adding 12 mL of mPEG. 2000 -DSPE (7 mg / mL) was added.

[0066] The resulting mixture was mixed at a low speed (approximately 60 rpm) for 5 minutes.

[0067] The final DTG concentration in the final DTG-CS suspension was approximately 1.64 mg / mL.

[0068] DTG-CS particle size and concentration The DTG-CS particle size and DTG concentration can be adjusted by changing the DTG:Gln ratio during the preparation process.

[0069] The following steps were taken to increase the DTG concentration.

[0070] When the above DTG-CS suspension (approximately 1.64 mg / mL) was left to stand overnight, DTG-CS particles settled at the bottom of the flask.

[0071] It removes approximately 92-93% of the liquid volume, excluding particles.

[0072] The particles are resuspended in 60 mL to achieve a final DTG concentration of approximately 20 mg / mL in DTG-CS.

[0073] The final product is stable at 25°C or 4°C with respect to particle size and drug concentration over a 6-8 month study period.

[0074] This method and formulation process are easily scalable and are intentionally designed as a simple, scalable process for producing injectable sterile drug products. No heating or cooling is required. Particle size was determined by dynamic light scattering (DLS) (the hydrodynamic size of particles determined by optically measuring particles undergoing Brownian motion with low scattering capacity). Particles had initial sizes ranging from approximately 200 to 700 nm (preferred nanoparticle size) or larger sizes ranging from approximately 1 to 2 microns, depending on the DTG:Gln ratio.

[0075] As described above, DTG-CS (dolutegravir:glutamine) particles were prepared by controlling the addition of glutamine to a dolutegravir solution. DSPE-mPEG2000 was included in the resulting product to avoid particle aggregation. DTG-CS (dolutegravir:glutamine) particles can be concentrated 10 to 100 times by sedimentation or centrifugation (low speed).

[0076] Effect of glutamine on DTG-Gln CS particle size In the formulation process, changing the glutamine concentration affects the particle size of DTG-Gln CS. The DTG:Gln ratio was adjusted to reduce the particle size of DTG-Gln CS. A direct relationship was observed between Gln concentration (%w / v) and particle size. Increasing the Gln concentration increased the particle size of DTG-Gln CS.

[0077] Figure 7 illustrates the effects of various concentrations of Gln on the DTG-Gln CS particle size.

[0078] DTG-Gln CS particle stability DTG-Gln CS particles prepared as described herein are stable at 25°C or 4°C with respect to particle size and drug concentration for a period of at least 6 to 8 months, and contain magnesium ions (Mg 2+ They are not inverted by ). DTG-Gln CS particles can be inverted by adding a high concentration of urea.

[0079] While exemplary embodiments have been illustrated and described, it will be understood that various modifications can be made without departing from the spirit and scope of the present invention.

Claims

1. Colloidal integrase chain transfer inhibitor particles containing an integrase chain transfer inhibitor and an amino acid.

2. The particle according to claim 1, wherein the integrase chain transfer inhibitor is selected from the group consisting of dolutegravir, bictegravir, cabotegravir, raltegravir, and elvitegravir.

3. The particle according to claim 1, wherein the integrase chain transfer inhibitor is selected from the group consisting of dolutegravir, bictegravir, and cabotegravir.

4. The particle according to claim 1, wherein the integrase chain transfer inhibitor is dolutegravir.

5. The particle according to claim 1, wherein the integrase chain transfer inhibitor is selected from the group consisting of raltegravir and elvitegravir.

6. The particle according to any one of claims 1 to 5, wherein the amino acid is glutamine or tryptophan.

7. The particle according to claim 1, wherein the integrase chain transfer inhibitor is dolutegravir (DTG) and the stabilizing amino acid is glutamine (Gln).

8. The particle according to claim 1, wherein the integrase chain transfer inhibitor is dolutegravir (DTG) and the stabilizing amino acid is tryptophan (Trp).

9. The particle according to claim 1, wherein the integrase chain transfer inhibitor is bictegravir (BIC) and the stabilizing amino acid is glutamine (Gln).

10. The particle according to claim 1, wherein the integrase chain transfer inhibitor is bictegravir (BIC) and the stabilizing amino acid is tryptophan (Trp).

11. An injectable pharmaceutical composition comprising a suspension of colloidal integrase chain transfer inhibitor particles according to any one of claims 1 to 10 in an aqueous carrier.

12. The composition according to claim 11, further comprising a biocompatible pharmaceutical excipient.

13. The composition according to claim 11 or 12, further comprising a pegylation modifier.

14. A method for treating or preventing HIV in a subject, comprising the step of administering a therapeutically effective amount of integrase chain transfer inhibitor particles according to any one of claims 1 to 10 or a composition according to any one of claims 11 to 13 to a subject in need thereof.

15. A method for treating a disease or condition that is preventable or treatable by administering an integrase chain transfer inhibitor, comprising the step of administering a therapeutically effective amount of integrase chain transfer inhibitor particles according to any one of claims 1 to 10 or a composition according to any one of claims 11 to 13 to a subject in need thereof.

16. The method according to claim 14 or 15, wherein the integrase chain transfer inhibitor particles or the composition are administered by subcutaneous or intramuscular injection.

17. The method according to claim 16, wherein the administration is at any interval between 30 and 365 days.

18. Integrase chain transfer inhibitor particles according to any one of claims 1 to 10 or a composition according to any one of claims 11 to 13, for use in the treatment or prevention of HIV infection in humans.

19. Integrate chain transfer inhibitor particles according to any one of claims 1 to 10 or a composition according to any one of claims 11 to 13, for the treatment or prevention of a disease or condition treatable by administering an integrase chain transfer inhibitor.

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