Drug preparations, methods of manufacturing the same, and uses

Abstract

A drug formulation comprising a ligand-drug conjugate, a lyophilized excipient, and a buffer is disclosed. By performing pH screening on the formulation and limiting the buffer and lyophilized excipient used therein, while simultaneously screening the lyophilization process, the effect of stably maintaining the appearance, moisture content, impurity content, and re-dissolution time of the formulation over the storage period is achieved.

Description

Technology Field

[0001] Priority and related applications

[0002] This application claims priority to a Chinese patent application filed with the National Intellectual Property Administration of China on September 8, 2021, with application number 202111051190.6 and title of invention “drug formulation, method of manufacturing and use thereof,” all of which are incorporated into this application by reference.

[0003] The present invention relates to the field of biopharmaceuticals, specifically to drug formulations, and specifically to drug formulations comprising a ligand-drug conjugate, a method of manufacturing the same, and uses. Background Technology

[0004] Various specific or overexpressed receptors, such as transferrin receptors (TFR), low-density lipoprotein receptors, folate receptors, uric acid kinase receptors, tumor necrosis factor receptors, intercellular adhesion molecules (ICAM), and integrin receptor LFA-1, exist on the surface of tumor cells and other cells with lesions (e.g., vascular endothelial cells, leukocytes, cerebral capillary endothelial cells). The ligands of these receptors can be classified into protein systems (ferritin, apolipoproteins, low-density lipoprotein receptor-associated protein 1, i.e., LRP1, etc.), polypeptide systems (insulin, SOR-C13, luteinizing hormone-releasing hormone LHRH, somatostatin SST-14, etc.), and small molecule systems (folate and analogs, sugar compounds). These ligands possess characteristics of excellent specificity, moderate affinity, and apparent biological effects when bound to receptors, allowing them to combine with therapeutic drugs to form ligand-drug conjugates (LDCs), which can significantly enhance drug targeting and efficacy while reducing toxicity. Ligand-drug conjugates possess endocytosis-mediating functions and can be applied to the treatment of tumors, immunomodulation, and cardiovascular diseases. Specifically, ① the combination of targeting and endocytosis structures allows any ligand to use a targeted polypeptide as a guide component, thereby broadening the target range of such drugs; ② the use of dual-targeting ligands enhances the affinity and targeting of the drug conjugate for lesion cells, enabling the possession of highly effective toxic drugs such as monomethylauristatin E (MMAE), thereby enhancing drug efficacy, widening the therapeutic window, and preventing drug side effects; and ③ the linker cannot release drug molecules outside the cell (intercellular substances, the circulatory system, etc.), ensuring drug stability during circulation, reducing drug toxicity, and providing the advantage of not having toxic effects on normal cells. After entering the target cell, the linker is cleaved to release a therapeutic drug molecule, thereby preventing the development of multidrug resistance (MDR).

[0005] In addition to formulating the conjugate in a manner suitable for administration to subjects, LDC formulations must be formulated in a way that maintains stability during storage and subsequent use. For example, if LDC is not properly formulated in a liquid, it tends to decompose, aggregate, or undergo unwanted chemical modifications within the liquid solution. The stability of LDC within the formulation is determined by the buffers, stabilizers, and surfactants used in the formulation.

[0006] There is a need in the industry for novel drug formulations containing LDCs that are sufficiently stable and suitable for administration to subjects; therefore, it is necessary to manufacture LDC formulations suitable for the treatment or prevention of diseases. [Prior Art][Patent] Chinese Published Patent Application CN 108135881 A (Published June 8, 2018) The problem to be solved

[0007] In light of the aforementioned industry demand for novel LDC formulations that are stable and suitable for administration to subjects, the present invention aims to provide a formulation comprising a ligand-drug conjugate that satisfies the requirement to maintain good stability over a storage period as an LDC formulation by limiting parameters such as pH value, buffer, lyophilized excipient, and lyophilized process, as well as a method for manufacturing the same and uses. means of solving the problem

[0008] In a first aspect, the present invention provides a drug formulation comprising an active drug and a selective pharmaceutically acceptable adjuvant.

[0009] Furthermore, in the above drug formulation, the active drug comprises a ligand-drug conjugate or a pharmaceutically acceptable salt thereof; and the pharmaceutically acceptable adjuvant comprises one or more of a lyophilized excipient and a buffer.

[0010] Furthermore, in the above drug formulation, the pharmaceutically acceptable adjuvant further comprises a solvent.

[0011] Furthermore, in the above drug formulation, the ligand-drug conjugate is a double ligand-drug conjugate; preferably, the double ligand-drug conjugate is a drug conjugate targeting folate receptors and TRPV6 receptors; more preferably, the double ligand-drug conjugate has the following structure,

[0012] ;

[0013] More preferably, the double ligand-drug conjugate has the following structure.

[0014]

[0015] Furthermore, in the above drug formulation, the solvent is water; preferably, the solvent is purified water; more preferably, the solvent is sterile water, distilled water, or deionized water; and more preferably, the sterile water is sterile water for injection, single distilled water, or double distilled water.

[0016] Furthermore, in the above drug formulation, the concentration of the active drug is 4.75 mg / mL to 6 mg / mL; preferably, the concentration of the active drug is 4.75 mg / mL, 5 mg / mL, 5.25 mg / mL, 5.5 mg / mL, 5.75 mg / mL, and 6 mg / mL.

[0017] Furthermore, the pH value of the above drug preparation is 6.5 to 8.5; preferably, the pH value of the above drug preparation is 6.9 to 8.0, for example, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0; more preferably, the pH value of the above drug preparation is 6.9 to 7.7, for example, 6.9, 7.0, 7.3, 7.5, 7.7.

[0018] Furthermore, in the above drug formulation, the lyophilized excipient comprises a polyol such as one or more of mannitol, sorbitol, inositol, and xylitol; preferably, the lyophilized excipient comprises mannitol.

[0019] Furthermore, in the above drug formulation, the freeze-dried excipient further comprises sugars such as one or more of monosaccharides, disaccharides, and polysaccharides; preferably, the mass ratio of the sugars to the polyol is 1:1 to 1:5; more preferably, the mass ratio of the sugars to the polyol is 1:2 to 1:4.

[0020] Preferably, in the above drug formulation, the monosaccharide comprises one or more of glucose and fructose; the disaccharide comprises one or more of sucrose, trehalose, maltose, and lactose; and the polysaccharide comprises one or more of cyclodextrin and dextran.

[0021] More preferably, in the above drug formulation, the lyophilized excipient comprises mannitol and sucrose; preferably, the mass ratio of sucrose to mannitol is 1:2 to 1:5, for example, 1:2, 1:2.4, 1:2.5, 1:4, 1:4.5, 1:4.7, 1:5; more preferably, the mass ratio of sucrose to mannitol is 1:4.

[0022] More preferably, in the above drug formulation, the lyophilized excipient comprises mannitol and trehalose; preferably, the mass ratio of trehalose to mannitol is 1:2 to 1:4; more preferably, the mass ratio of trehalose to mannitol is 1:4.

[0023] Furthermore, in the above drug formulation, the buffer comprises tromethamine and an acidic substance; the mass ratio of tromethamine to the active drug is 1:4 to 1:10, preferably 1:5 to 1:8.5.

[0024] Preferably, in the above drug formulation, the acidic substance comprises one or more of hydrochloric acid, acetic acid, sodium dihydrogen phosphate, and tromethamine hydrochloride.

[0025] More preferably, in the above drug formulation, the buffer comprises tromethamine and hydrochloric acid; preferably, the mass ratio of the tromethamine to the active drug is 1:5 to 1:8.5, for example, 1:5, 1:5.5, 1:6, 1:6.5, 1:7, 1:8, 1:8.5; more preferably, the ratio of the tromethamine to the active drug is 1:6.5.

[0026] Furthermore, in the above drug formulation, the drug formulation comprises 4.75 to 6 mg of a ligand-drug conjugate or a pharmaceutically acceptable salt thereof, 25 to 55 mg of mannitol, 0 to 55 mg of sucrose, 0.5 to 1.25 mg of tromethamine, hydrochloric acid, and the remainder of a solvent per 1 mL; wherein the pH of the drug formulation is 6.5 to 8.5, preferably 6.9 to 8.0; for example, the drug formulation comprises 5 mg of a ligand-drug conjugate or a pharmaceutically acceptable salt thereof, 40 mg of mannitol, 10 mg of sucrose, 0.765 mg of tromethamine, hydrochloric acid, and the remainder of water per 1 mL; wherein the pH value of the drug formulation is 6.5 to 8.5, preferably 6.9 to 8.0.

[0027] Furthermore, the above drug preparation is sterilized.

[0028] Furthermore, the above drug formulation is stable upon freezing and thawing.

[0029] In a second aspect, the present invention provides a method for preparing the drug formulation, comprising the step of taking a prescribed amount of an active drug and an optional pharmaceutically acceptable adjuvant and mixing them uniformly.

[0030] Furthermore, the above manufacturing method comprises the steps of taking a prescribed amount of a ligand-drug conjugate or a pharmaceutically acceptable salt thereof, a lyophilized excipient, a buffer, and a solvent, dissolving the ligand-drug conjugate or the pharmaceutically acceptable salt thereof and the lyophilized excipient using the solvent, adjusting the pH value to 6.9 to 7.7 using the buffer, mixing uniformly, and obtaining by filtration.

[0031] In a third aspect, the present invention provides a freeze-dried formulation characterized by being prepared by freeze-drying the above drug formulation.

[0032] In a fourth aspect, the present invention provides a method for preparing the freeze-dried formulation, comprising: (i) freezing the drug formulation at -45°C; (ii) annealing the drug formulation at -25°C; (iii) refreezing the drug formulation at -45°C; (iv) first drying the drug formulation at -15°C; and (v) second drying the drug formulation at 25°C.

[0033] Furthermore, in the above manufacturing method, the first drying time is 800 to 3000 min; and the second drying time is 300 to 600 min.

[0034] In a fifth aspect, the present invention provides a liquid formulation formed by reconstituting the freeze-dried formulation with water.

[0035] Furthermore, in the above liquid formulation, the water comprises one or more of distilled water, pure water, and sterile water; preferably, the pH value of the freeze-dried formulation reconstituted with water is 6.5 to 8.5.

[0036] In a sixth aspect, the present invention provides a method for preparing the liquid formulation, which includes the step of mixing the freeze-dried formulation with sterile water.

[0037] In a seventh aspect, the present invention provides a drug delivery device comprising one of the drug formulation, the lyophilized formulation, or the liquid formulation.

[0038] In an eighth aspect, the present invention provides a pre-filled syringe comprising one of the drug formulation, the lyophilized formulation, or the liquid formulation, and preferably used for intravenous injection or intramuscular injection.

[0039] In a ninth aspect, the present invention provides the use of the drug formulation, the lyophilized formulation, and the liquid formulation in the manufacture of a delivery device or pre-filled syringe or drug for enhancing the immune effector cell response of a subject and / or reducing immunosuppression.

[0040] In a tenth aspect, the present invention provides the use of the drug formulation, the lyophilized formulation, and the liquid formulation in the manufacture of a delivery device or pre-filled syringe or drug for treating or preventing cancer, immune disease, cardiovascular disease, metabolic disease, and neurological disease of a subject.

[0041] Furthermore, in the above use, the cancer includes one or more of breast cancer, lung cancer, prostate cancer, kidney cancer, leukemia, ovarian cancer, stomach cancer, uterine cancer, endometrial cancer, liver cancer, colon cancer, thyroid cancer, pancreatic cancer, colorectal cancer, esophageal cancer, skin cancer, lymphoma, and multiple myeloma; the immune disease includes one or more of connective tissue disease, systemic sclerosis, rheumatoid arthritis, and systemic lupus erythematosus; the cardiovascular disease includes one or more of angina pectoris, myocardial infarction, stroke, heart attack, hypertensive heart disease, rheumatic heart disease, cardiomyopathy, cardiac arrhythmia, and congenital heart disease; the metabolic disease includes one or more of diabetes mellitus, gout, obesity, hypoglycemia, hyperglycemia, and dyslipidemia; and the neurological disease includes one or more of Alzheimer's disease, Parkinson's disease, Huntington's disease, head injury, multiple sclerosis, vertigo, coma, and epilepsy. Effects of the invention

[0042] The present invention performs pH screening on a formulation comprising a ligand-drug conjugate, limits the buffer and freeze-drying excipients used therein, and simultaneously screens the freeze-drying process, thereby achieving the effect of stably maintaining the appearance, moisture content, impurity content, and re-dissolution time of the formulation during storage. Specific details for implementing the invention

[0043] Embodiments of the present invention are described below, but the present invention is not limited thereto. The present invention is not limited to each composition described below, and various modifications are possible within the scope of the claimed invention. Furthermore, embodiments and examples obtained by appropriately combining the technical solutions disclosed in each of the different embodiments and examples are also included in the present invention. Additionally, all documents described in this specification are cited by reference in this specification.

[0044] Unless otherwise defined, technical and scientific terms used herein have the same meaning as generally understood by a person skilled in the art to which the present invention pertains.

[0045] In this specification, a numeric range expressed using “numerical value A to numerical value B” or “numerical value A - numerical value B” means a range including endpoint numerical values ​​A and B.

[0046] In this specification, “%” indicates a mass or weight percentage content, i.e., “mass%” or “weight%”, unless otherwise specified.

[0047] In this specification, “room temperature” or “room temperature” means an ambient temperature of 25°C.

[0048] In this specification, the meaning expressed using “…may” includes both the meaning of performing a specific process and the meaning of not performing a specific process. In this specification, “optional” or “optionally” means that an event or situation to be described below may or may not occur, and said description includes situations in which said event occurs and situations in which said event does not occur.

[0049] In this specification, references to “some specific / preferred embodiments,” “other specific / preferred embodiments,” “some specific / preferred technical solutions,” “other specific / preferred technical solutions,” etc., mean that specific elements (e.g., features, structures, properties, and / or characteristics) related to the described embodiments are included in at least one embodiment described in this specification and may or may not be present in other embodiments. Furthermore, it should be understood that said elements may be combined in any appropriate manner in various embodiments.

[0050] In the specification and claims of the present invention, the term “inclusion” and any variation thereof are intended to encompass non-exclusive inclusion.

[0051] In the present invention, “pharmaceuticalally acceptable” means suitable for contact with human and other animal cells without excessive toxicity, irritation, allergic reactions, etc., within the scope of reasonable medical judgment, and proportional to a reasonable benefit / risk ratio.

[0052] In the present invention, “pharmaceuticalally acceptable salt” refers to the relatively non-toxic inorganic and organic acid addition salts and alkali addition salts of the conjugate compound of the present application. Representative acid addition salts include hydrobromide, hydrochloride, sulfate, bisulfate, phosphate, nitrate, acetate, oxalate, valerate, oleate, palmitate, stearate, laurate, borate, benzoate, lactate, tosylate, citrate, maleate, fumarate, succinate, tartrate, naphthenate, mesylate, glucoheptonate, lactobionate, sulfamate, malonate, salicylate, propionate, methylene-bis-b-hydroxynaphtoate, gentisate, isethionate, di-p-toluoil tartrate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, cyclamate, and quinateslaurylsulphonate. The base addition salt comprises pharmaceutically acceptable metal and amine salts. Suitable metal salts include sodium, potassium, calcium, barium, zinc, magnesium, and aluminum salts. In some embodiments, sodium and potassium salts are preferred. Suitable inorganic base addition salts are prepared from metal salts, said metal salts including, for example, sodium hydroxide, sodium hydroxide, potassium hydroxide, calcium hydroxide, aluminum hydroxide, lithium hydroxide, magnesium hydroxide, and zinc hydroxide.Suitable amine base addition salts are prepared from amines having sufficient basicity to form a stable salt, and preferably include amines frequently used in pharmacochemistry such as ammonia, ethylenediamine, N-methyl-glucarmine, lysine, arginine, ornithine, choline, N,N'-dibenzylethylenediamine, chloroprocaine, diethanolamine, procaine, N-benzylphenethylamine, diethylamine, piperazine, tri(hydroxymethyl)-aminomethane, tetramethylammonium hydroxide, triethylamine, dibenzylamine, ephenamine, dehydroaviethylamine, N-ethylpiperadine, benzylamine, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, ethylamine, basic amino acids (e.g., lysine and arginine), and dicyclohexylamine, as they have low toxicity and are acceptable for medical use.

[0053] In the present invention, the “ligand-drug conjugate” or “ligand-drug conjugate” (LDC) links a biologically active drug to a ligand through a chemical linkage, and the ligand acts as a carrier to deliver a small molecule drug to a target cell, wherein the ligand may be a polypeptide or a small molecule. Preferably, the ligand-drug conjugate is a double ligand-drug conjugate, and preferably, the double ligand-drug conjugate is a drug conjugate targeting folate receptors and TRPV6 receptors. More preferably, the ligand-drug conjugate of the present invention is the ligand-drug conjugate described in the patent application with application number 201680045855.3 and title of invention “Multiple ligand-drug conjugate and uses thereof,” and more preferably, the ligand conjugate compound is the double ligand conjugate LDC10B, wherein the ligands target the TRPV6 receptor and the folate receptor, respectively, and the loading drug is MMAE, and the structure thereof is as follows.

[0054]

[0055] More preferably, the double ligand-drug conjugate has the following structure.

[0056]

[0057] In the present invention, a “buffer” is a reagent that maintains the pH value of a solution within an acceptable range. In some embodiments, the buffer used in the formulation of the present invention controls the pH of the formulation of the present invention to a pH range of about 6.5 to 8.5, and can control the pH value to, for example, about 6.9 to 8.0. In some specific embodiments, the formulation of the present invention has pH values ​​of about 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, and 8.0.

[0058] In the present invention, “polyol” refers to a large class of alcohols containing two or more hydroxyl groups in the molecule.

[0059] In the present invention, “reconstitution” means dissolving and / or suspending a solid formulation (e.g., a lyophilized formulation) in a physiologically acceptable solution.

[0060] In the present invention, “approximately” means that the limited numerical value is approximate, and that the specific numerical value in the relevant data does not need to be too precise.

[0061] In the present invention, “preparation” refers to a pharmaceutical product manufactured according to specific formulation requirements and ultimately provided to a drug target for use.

[0062] In the present invention, “freeze-dried formulation” refers to a sterile injectable solid prepared by freeze-drying.

[0063] In the present invention, the “freeze-drying excipient,” also referred to as a freeze-drying protective agent, refers to an additive added during the freeze-drying and storage process of food, pharmaceuticals, or organisms, and its purpose is to maintain stability and activity even when affected by various factors (e.g., chemical composition, freezing rate, freeze-drying stress, glass transition temperature, residual moisture of the dry solid, temperature and humidity of the storage environment, etc.).

[0064] In the present invention, “stable formulation” refers to a formulation in which the moisture content, appearance, pH value, impurity content, re-dissolving properties, and shape do not fundamentally change or change within a pharmaceutically acceptable range during the manufacturing and storage process of the product.

[0065] In the present invention, “HPLC” refers to High Performance Liquid Chromatography, which is an important field of chromatography. A liquid is used as the mobile phase, and a high-pressure injection system is used to pump the mobile phase, such as a single solvent with different polarities or a mixed solvent of different ratios, or a buffer, into a chromatography column equipped with a stationary phase. After separating each component within the column, the components are introduced into a detector to perform sample analysis.

[0066] [Drug formulation containing LDC and method of manufacturing the same]

[0067] The present invention provides a drug formulation that may include an active drug and a selective pharmaceutically acceptable adjuvant.

[0068] In one embodiment, the drug formulation may comprise a ligand-drug conjugate, a lyophilized excipient, and a buffer.

[0069] In one embodiment, the drug formulation may comprise a ligand-drug conjugate, a lyophilized excipient, a buffer, and a solvent.

[0070] In a preferred embodiment, the ligand-drug conjugate in the drug formulation may be a dual ligand-drug conjugate targeting folate receptors and TRPV6 receptors.

[0071] In one embodiment, the solvent in the drug formulation is water.

[0072] In a preferred embodiment, the solvent in the drug formulation is purified water.

[0073] In another preferred embodiment, the solvent in the drug formulation is sterile water, distilled water, or deionized water.

[0074] In a more preferred embodiment, the solvent in the drug formulation is sterile water for injection, single distilled water, or double distilled water.

[0075] In one embodiment, the active drug concentration in the drug formulation is 4.75 mg / mL to 6 mg / mL, for example 4.75 mg / mL, 5 mg / mL, 5.25 mg / mL, 5.5 mg / mL, 5.75 mg / mL, and 6 mg / mL.

[0076] In a preferred embodiment, the active drug concentration of the drug formulation is 5 mg / mL.

[0077] In one embodiment, the pH value of the drug formulation may be 6.5 to 8.5.

[0078] In a preferred embodiment, the pH value of the drug formulation may be 6.9 to 8.0, for example, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0.

[0079] In a more preferred embodiment, the pH value of the drug formulation may be 6.9 to 7.7, for example, 6.9, 7.0, 7.3, 7.5, and 7.7.

[0080] In one embodiment, the lyophilized excipient in the drug formulation may include a polyol such as one or more of mannitol, sorbitol, inositol, and xylitol.

[0081] In a preferred embodiment, the lyophilized excipient in the drug formulation comprises mannitol.

[0082] In one embodiment, the lyophilized excipient in the drug formulation comprises a polyol and a sugar, and includes, for example, one or more of a monosaccharide, a disaccharide, and a polysaccharide.

[0083] Furthermore, the monosaccharides in the lyophilized excipients of the above drug formulation include one or more of glucose and fructose; the disaccharides include one or more of sucrose, trehalose, maltose, and lactose; and the polysaccharides include one or more of cyclodextrin and dextran.

[0084] Furthermore, the mass ratio of the sugars and the polyol is 1:1 to 1:5; preferably, the mass ratio of the sugars and the polyol is 1:2 to 1:4.

[0085] In a preferred embodiment, the lyophilized excipient in the drug formulation may include mannitol and sucrose, and preferably, the mass ratio of sucrose to mannitol is 1:2 to 1:5, for example, 1:2, 1:2.4, 1:2.5, 1:4, 1:4.5, 1:4.7, 1:5.

[0086] In a more preferred embodiment, the lyophilized excipient in the drug formulation may include mannitol and sucrose, and preferably, the mass ratio of sucrose to mannitol is 1:4.

[0087] In another preferred embodiment, the lyophilized excipient in the drug formulation may include mannitol and trehalose, and preferably, the mass ratio of trehalose to mannitol is 1:2 to 1:4.

[0088] In another more preferred embodiment, the lyophilized excipient in the drug formulation may include mannitol and trehalose, and preferably, the mass ratio of trehalose to mannitol is 1:4.

[0089] In one embodiment, the buffer in the drug formulation may include tromethamine and an acidic substance, and the mass ratio of tromethamine to the active drug is 1:4 to 1:10, preferably 1:5 to 1:8.5.

[0090] Furthermore, the acidic substance included in the buffer within the above drug formulation may be selected from one or more of hydrochloric acid, acetic acid, sodium dihydrogen phosphate, and tromethamine hydrochloride.

[0091] In a preferred embodiment, the buffer in the drug formulation may comprise tromethamine and hydrochloric acid, and preferably, the mass ratio of tromethamine to the active drug is 1:5 to 1:8.5, for example, 1:5, 1:5.5, 1:6, 1:6.5, 1:7, 1:8, 1:8.5.

[0092] In a more preferred embodiment, the buffer in the drug formulation may include tromethamine and hydrochloric acid, and preferably, the ratio of tromethamine to the active drug is 1:6.5.

[0093] In another preferred embodiment, the buffer in the drug formulation may include tromethamine and acetic acid, and the ratio of tromethamine to the active drug is 1:6.5.

[0094] In another preferred embodiment, the buffer in the drug formulation may include tromethamine and sodium dihydrogen phosphate, and the ratio of tromethamine to the active drug is 1:6.5.

[0095] In another preferred embodiment, the buffer in the drug formulation may include tromethamine and tromethamine hydrochloride, and the ratio of tromethamine to the active drug is 1:6.5.

[0096] In one embodiment, the drug formulation comprises 4.75 to 6 mg of a ligand-drug conjugate or a pharmaceutically acceptable salt thereof per 1 mL, 25 to 55 mg of mannitol, 0 to 55 mg of sucrose, 0.5 to 1.25 mg of tromethamine, hydrochloric acid, and the remainder being a solvent; wherein the pH value of the drug formulation is 6.5 to 8.5, preferably 6.9 to 8.0.

[0097] In a preferred embodiment, the drug formulation comprises 5 mg of a ligand-drug conjugate or a pharmaceutically acceptable salt thereof per 1 mL, 40 mg of mannitol, 10 mg of sucrose, 0.765 mg of tromethamine, hydrochloric acid, and the remainder being water; wherein the pH value of the drug formulation is 6.5 to 8.5, preferably 6.9 to 8.0.

[0098] In one embodiment, the drug formulation is sterilized.

[0099] In one embodiment, the drug formulation is stable during freezing and thawing.

[0100] In one embodiment, the method for preparing the drug formulation comprises the step of taking a prescribed amount of an active drug and an optional pharmaceutically acceptable adjuvant, and uniformly mixing them to obtain the drug formulation.

[0101] In a preferred embodiment, the method for preparing the drug formulation comprises the steps of taking a prescribed amount of a ligand-drug conjugate or a pharmaceutically acceptable salt thereof, a lyophilized excipient, a buffer, and a solvent, dissolving the ligand-drug conjugate or the pharmaceutically acceptable salt thereof and the lyophilized excipient using the solvent, adjusting the pH value to 6.9 to 7.7 using the buffer, mixing uniformly, and filtering to obtain the result.

[0102] [Freeze-dried formulation and method of manufacturing the same]

[0103] The present invention provides a freeze-dried formulation prepared by freeze-drying the drug formulation of the present invention.

[0104] The present invention further provides a method for manufacturing the freeze-dried formulation comprising the step of obtaining the drug formulation of the present invention by freeze-drying.

[0105] In one embodiment, the manufacturing method comprises the step of freezing the drug preparation at -45°C; the step of annealing the drug preparation at -25°C; the step of refreezing the drug preparation at -45°C; the step of first drying the drug preparation at -15°C; and the step of secondarily drying the drug preparation at 25°C.

[0106] In a preferred embodiment, the manufacturing method comprises the steps of: freezing a drug preparation at -45°C; annealing a drug preparation at -25°C; refreezing a drug preparation at -45°C; first drying a drug preparation at -15°C for 800 to 3000 min; and second drying a drug preparation at 25°C for 300 to 600 min.

[0107] In a more preferred embodiment, the manufacturing method comprises: a freezing step of placing a prescription solution into a freeze dryer box and cooling it down to -45°C at atmospheric pressure and maintaining it for 120 minutes; an annealing step of cooling it down to -25°C at atmospheric pressure and maintaining it for 180 minutes; a freezing step of cooling it down to -45°C at atmospheric pressure and maintaining it for 120 minutes; a pre-vacuuming step of changing the atmospheric pressure of the box to 0.2 mbar; a first drying step of cooling it down to -15°C at 0.13 mbar and maintaining it for 800 to 3000 minutes; and a second drying step of cooling it down to 25°C at 0.13 mbar and maintaining it for 300 to 600 minutes.

[0108] In a more preferred embodiment, the manufacturing method comprises: a freezing step of placing a prescription solution into a freeze dryer box, cooling the temperature to -45°C at atmospheric pressure and maintaining it for 120 minutes; an annealing step of cooling the temperature to -25°C at atmospheric pressure and maintaining it for 180 minutes; a freezing step of cooling the temperature to -45°C at atmospheric pressure and maintaining it for 120 minutes; a pre-vacuuming step of changing the atmospheric pressure of the box to 0.2 mbar; a first drying step of cooling the temperature to -15°C at 0.13 mbar and maintaining it for 800 minutes; and a second drying step of cooling the temperature to 25°C at 0.13 mbar and maintaining it for 600 minutes.

[0109] [Liquid formulation and method of manufacturing the same]

[0110] The present invention provides a liquid formulation prepared by reconstituting the freeze-dried formulation of the present invention with water.

[0111] The present invention further provides a method for preparing the liquid formulation, comprising the step of mixing the freeze-dried formulation of the present invention with water.

[0112] In one embodiment, in the above manufacturing method, the reconstitution is completed using distilled water, pure water, or sterile water.

[0113] In one embodiment, the pH value of the freeze-dried formulation reconstituted with water is 6.5 to 8.5.

[0114] [device]

[0115] The present invention provides a drug delivery device comprising one of the above drug formulation, the above freeze-dried formulation, or the above liquid formulation.

[0116] The present invention further provides a pre-filled syringe comprising one of the above drug formulation, the above lyophilized formulation, or the above liquid formulation.

[0117] [Pharmacological Use]

[0118] The present invention provides the use of the drug formulation, the lyophilized formulation, or the liquid formulation comprising an active drug in the manufacture of a delivery device or a pre-filled syringe or a drug.

[0119] In one embodiment, the delivery device, pre-filled syringe, or drug in the above application is used to enhance the immune effector cell response of the subject.

[0120] In one embodiment, the delivery device or pre-filled syringe or drug in the above application is used to reduce immunosuppression in the subject.

[0121] In one embodiment, the delivery device, pre-filled syringe, or drug used for the above application is used to treat or prevent cancer in a subject.

[0122] Furthermore, in the above application, cancer includes one or more of breast cancer, lung cancer, prostate cancer, kidney cancer, leukemia, ovarian cancer, stomach cancer, uterine cancer, endometrial cancer, liver cancer, colon cancer, thyroid cancer, pancreatic cancer, colorectal cancer, esophageal cancer, skin cancer, lymphoma, and multiple myeloma.

[0123] In one embodiment, the delivery device, pre-filled syringe, or drug used for the above application is used to treat or prevent immune diseases in a subject.

[0124] Furthermore, in the above applications, immune diseases include one or more of connective tissue diseases, systemic sclerosis, rheumatoid arthritis, and systemic lupus erythematosus.

[0125] In one embodiment, the delivery device, pre-filled syringe, or drug used for the above application is used to treat or prevent cardiovascular disease in a subject.

[0126] Furthermore, in the above applications, cardiovascular disease includes one or more of angina pectoris, myocardial infarction, stroke, heart attack, hypertensive heart disease, rheumatic heart disease, cardiomyopathy, cardiac arrhythmia, and congenital heart disease.

[0127] In one embodiment, the delivery device, pre-filled syringe, or drug used for the above application is used to treat or prevent metabolic diseases of the subject.

[0128] Furthermore, in the above applications, metabolic diseases include one or more of diabetes mellitus, gout, obesity, hypoglycemia, hyperglycemia, and dyslipidemia.

[0129] In one embodiment, the delivery device, pre-filled syringe, or drug used for the above application is used to treat or prevent neurological diseases in a subject.

[0130] Furthermore, in the above applications, neurological diseases include one or more of Alzheimer's disease, Parkinson's disease, Huntington's disease, head injury, multiple sclerosis, vertigo, coma, and epilepsy.

[0131] Unless otherwise specified, the apparatus, reagents, materials, etc. used in the present invention may be obtained through ordinary commercial means. The technical solution of the present invention will be further explained below with reference to specific embodiments.

[0132] Example 1

[0133] This is to test the stability of a formulation containing LDC10B at pH 6.9 to 8.0. A total of five pH values ​​were designed, namely, pH values ​​of 6.9, 7.2, 7.5, 7.8, and 8.0, respectively.

[0134] 1.1 Experimental Steps

[0135] Aqueous solutions containing LDC10B (5 mg / mL), mannitol (40 mg / mL), sucrose (10 mg / mL), and tromethamine (0.765 mg / mL) were prepared, and the pH was adjusted with dilute hydrochloric acid to 6.9, 7.2, 7.5, 7.8, and 8.0, respectively; the solutions were filtered, dispensed into vials, plugged, and the lids closed. The stability of each sample was tested for 12 hours at room temperature.

[0136] 1.2 Experimental Plan

[0137] The specific plan is as shown in the table below.

[0138]

[0139] 1.3 Experimental Results

[0140]

[0141] After being left at room temperature for different times at pH 6.9, 7.2, 7.5, 7.8, and 8.0, the relevant substances in each sample were measured by HPLC. As can be seen from this, when tested for 12 h under room temperature conditions, the total impurities in all samples at each pH value increased, and compared to the relevant substances in the samples at 0 h, the total impurities in the samples at pH 6.9, 7.2, 7.5, 7.8, and 8.0 increased by 0.17%, 0.37%, 0.58%, 0.68%, and 1.28%, respectively.

[0142] The above experimental results show that the total impurities are all within the acceptable range between pH 6.9 and 8.0.

[0143] 1.4 Prepare the sample in the final formulation form (freeze-dried form) and check the pH.

[0144] 1.4.1 Experimental Phase

[0145] Aqueous solutions containing LDC10B (5 mg / mL), mannitol (40 mg / mL), sucrose (10 mg / mL), and tromethamine (0.765 mg / mL) were prepared, and the pH was adjusted to 6.9, 7.3, and 7.7, respectively, using dilute hydrochloric acid; the solutions were filtered, dispensed into vials, sealed in half, freeze-dried, and then closed. Stability was tested for each sample for one month at 30°C.

[0146] 1.4.2 Experimental Plan

[0147] The specific plan is as shown in the table below.

[0148]

[0149] 1.4.3 Experimental Results

[0150]

[0151] The experimental results are as shown in the table above, and as can be seen from the data, the quality of the freeze-dried formulation containing LDC10B is stable within the pH range of 6.9 to 7.7.

[0152] Example 2

[0153] To study the effects of tromethamine buffering, phosphate buffering, and acetate buffering on the stability of formulations containing LDC10B, a total of four formulations were designed, and detailed formulation information is shown in the table below.

[0154]

[0155] 2.1 Experimental Phase

[0156] Aqueous solutions of each of the above formulations were prepared, filtered, divided into vials, sealed in half, freeze-dried, and the lids closed. Each sample was tested for stability under conditions of 40°C.

[0157] 2.2 Experimental Plan

[0158] The specific measures are as shown in the table below, and the detection indicators are appearance and related substances.

[0159]

[0160] 2.3 Experimental Results

[0161]

[0162] The above experimental results show that the change in total impurities in each formulation under 40℃ conditions is all within an acceptable range.

[0163] Example 3

[0164] The amount of tromethamine used in the obtained buffer system (0.615 ~ 1.00 mg / mL) was screened and optimized to obtain better product quality.

[0165]

[0166] 3.1 Experimental Phase

[0167] Aqueous solutions of each of the above formulations were prepared, filtered, divided into vials, sealed in half, freeze-dried, and the lids closed. The stability of each sample was tested under conditions of 2–8°C.

[0168] 3.2 Experimental Results

[0169]

[0170] The results show that when the tromethamine usage is 0.615 mg / mL to 1.00 mg / mL, there are no significant differences in product re-dissolution time, properties and shape, moisture, and related substances, and that 0.615 mg / mL to 1.00 mg / mL is an acceptable usage range.

[0171] Example 4

[0172] To study the effects of mannitol as a polyol and sucrose and trehalose as sugars on formulations containing LDC10B, four formulations were designed, and detailed formulation information is as shown in the table below.

[0173]

[0174] 4.1 Experimental Phase

[0175] Aqueous solutions of each of the above prescriptions were prepared, filtered, divided into vials, sealed in half, freeze-dried, and the lids closed. After freeze-drying and removing from the box, the products of prescriptions 9 and 10 shrank, but this phenomenon did not occur in the other prescriptions.

[0176] 4.2 Experimental Plan

[0177] The specific measures are as shown in the table below, and the detection indicators are appearance, moisture, and related substances.

[0178]

[0179] 4.3 Experimental Results

[0180]

[0181] Note: “NA” means there is no relevant data.

[0182] As can be seen from the above experimental results, when sucrose (Formula 9) and trehalose (Formula 10) are used as fillers, the product shrinks, and after adding mannitol, a freeze-dried preparation with acceptable appearance, moisture, and quality can be obtained, so mannitol and sucrose can be selected, or mannitol and trehalose can be used together as excipients.

[0183] Example 5

[0184] In the obtained excipient system, better product quality was achieved by screening and optimizing the usage of mannitol and sucrose.

[0185]

[0186] 5.1 Experimental Phase

[0187] Aqueous solutions of each of the above formulations were prepared, filtered, divided into vials, sealed in half, freeze-dried, and the lids closed. Each sample was tested for stability under conditions of 40°C.

[0188] 5.2 Experimental Results

[0189]

[0190] As can be seen from the results, when mannitol is used alone, the impurities in the product increase significantly, and when sucrose and mannitol are used together as excipients, the increase in impurities is significantly reduced; therefore, the amount of excipients used in this product is 20 mg to 33.5 mg of sucrose and 80 mg to 93.5 mg of mannitol.

[0191] Example 6

[0192] 6.1 Process Selection

[0193]

[0194] 6.2 Experimental Results

[0195]

[0196] Through all processes, products were obtained that had a good appearance, acceptable moisture content, and met clinical use requirements for re-dissolution time and pH values. In the freeze-drying process, the acceptable freezing temperature is -45°C and undergoes annealing at -25°C; the first drying temperature is -15°C and the time is 800 min to 3000 min; the second drying temperature is 25°C and the time is 300 min to 600 min.

[0197] Example 7

[0198] The stability of the final prescription was tested by performing a prescription process verification experiment in conjunction with the results of the prescription screening experiment.

[0199] Prescription information is as shown in the table below.

[0200]

[0201] Process information is as shown in the table below.

[0202]

[0203] The experimental plan is as shown in the table below.

[0204]

[0205] The experimental results are as follows.

[0206]

[0207] As can be seen from the results, when the product manufactured according to the established formula and process was exposed to 25°C for 6 months and to a high temperature of 40°C for 1 month, the changes in appearance, moisture (<5%), re-dissolving pH (6.9 to 7.7) and related substances (total impurities <5%) were all within an acceptable range.

Claims

Claim 1 A drug formulation comprising an active drug and a pharmaceutically acceptable adjuvant, wherein the active drug comprises a ligand-drug conjugate or a pharmaceutically acceptable salt thereof; the pharmaceutically acceptable adjuvant comprises a lyophilized excipient, wherein the lyophilized excipient comprises a polyol and a sugar; wherein the polyol is mannitol and the sugar is selected from sucrose and trehalose; wherein the mass ratio of the sugar to the polyol is 1:1 to 1:5; wherein the ligand-drug conjugate is a double ligand-drug conjugate; and wherein the double ligand-drug conjugate is a drug conjugate targeting a folate receptor and a TRPV6 receptor and has the following structure: . Claim 2 A drug formulation according to claim 1, characterized in that the double ligand-drug conjugate has the following structure: . Claim 3 A drug formulation according to claim 1, characterized in that the pharmaceutically acceptable adjuvant further comprises one or more of a buffer and a solvent. Claim 4 A drug formulation characterized in that, in paragraph 3, the solvent is water. Claim 5 A drug formulation according to claim 3, characterized in that the concentration of the active drug is 4.75 mg / mL to 6 mg / mL. Claim 6 A drug formulation according to claim 5, characterized in that the concentration of the active drug is 4.75 mg / mL, 5 mg / mL, 5.25 mg / mL, 5.5 mg / mL, 5.75 mg / mL, or 6 mg / mL. Claim 7 A drug formulation according to claim 1, characterized in that the pH value of the drug formulation is 6.5 to 8.

5. Claim 8 A drug formulation according to claim 7, characterized in that the pH value of the drug formulation is 6.9 to 7.

7. Claim 9 A drug formulation according to claim 1, wherein the freeze-dried excipient comprises mannitol and trehalose and the mass ratio of trehalose to mannitol is 1:2 to 1:4; or wherein the freeze-dried excipient comprises mannitol and sucrose and the mass ratio of sucrose to mannitol is 1:2 to 1:

5. Claim 10 A drug formulation according to claim 9, characterized in that the mass ratio of sucrose to mannitol is 1:2, 1:2.4, 1:2.5, 1:4, 1:4.5, 1:4.7, or 1:

5. Claim 11 A drug formulation according to claim 3, wherein the buffer comprises tromethamine and an acidic substance; and wherein the mass ratio of tromethamine to the active drug is 1:4 to 1:

10. Claim 12 A drug formulation according to claim 11, characterized in that the mass ratio of the tromethamine to the active drug is 1:5 to 1:8.

5. Claim 13 A drug formulation according to claim 11, characterized in that the acidic substance comprises one or more of hydrochloric acid, acetic acid, sodium dihydrogen phosphate, and tromethamine hydrochloride. Claim 14 A drug formulation according to claim 11, characterized in that the buffer comprises tromethamine and hydrochloric acid. Claim 15 A drug formulation according to claim 14, characterized in that the mass ratio of the tromethamine to the active drug is 1:5 to 1:8.

5. Claim 16 A drug formulation according to claim 1, wherein the pharmaceutically acceptable adjuvant further comprises one or more of a buffer and a solvent; wherein the polyol is mannitol, the sugar is sucrose, and the buffer comprises tromethamine and hydrochloric acid; and wherein the drug formulation comprises 4.75 to 6 mg of a ligand-drug conjugate or a pharmaceutically acceptable salt thereof per 1 mL, 25 to 55 mg of mannitol, 0 to 55 mg of sucrose, 0.5 to 1.25 mg of tromethamine, hydrochloric acid such that the pH value of the drug formulation is 6.5 to 8.5, and the remainder being a solvent. Claim 17 In claim 16, the drug formulation is characterized by comprising 5 mg of a ligand-drug conjugate or a pharmaceutically acceptable salt thereof per 1 mL, 40 mg of mannitol, 10 mg of sucrose, 0.765 mg of tromethamine, hydrochloric acid such that the pH value of the drug formulation is 6.5 to 8.5, and the remainder being water. Claim 18 A method for manufacturing a drug formulation according to paragraph 3, wherein the manufacturing method comprises the steps of: taking a prescribed amount of a ligand-drug conjugate or a pharmaceutically acceptable salt thereof, a lyophilized excipient, a buffer, and a solvent; dissolving the ligand-drug conjugate or the pharmaceutically acceptable salt thereof and the lyophilized excipient using the solvent; adjusting the pH value to 6.9 to 7.7 using the buffer; mixing uniformly; and filtering to obtain a drug formulation. Claim 19 A freeze-dried preparation, wherein the freeze-dried preparation is characterized by being prepared by freeze-drying a drug preparation according to any one of claims 1 to 17. Claim 20 A method for manufacturing a freeze-dried preparation according to claim 19, wherein the manufacturing method comprises: (i) freezing the drug preparation at -45℃; (ii) annealing the drug preparation at -25℃; (iii) refreezing the drug preparation at -45℃; (iv) first drying the drug preparation at -15℃; and (v) second drying the drug preparation at 25℃. Claim 21 A manufacturing method according to claim 20, characterized in that the first drying time is 800 to 3000 min; and the second drying time is 300 to 600 min. Claim 22 A drug delivery device comprising a drug formulation according to any one of claims 1 to 17. Claim 23 A pre-filled syringe comprising a drug formulation according to any one of claims 1 to 17. Claim 24 A drug preparation according to any one of claims 1 to 17 for use in treating or preventing cancer, immune diseases, cardiovascular diseases, metabolic diseases, and neurological diseases of a subject. Claim 25 A drug according to claim 24, wherein the above cancer includes one or more of breast cancer, lung cancer, prostate cancer, kidney cancer, leukemia, ovarian cancer, stomach cancer, uterine cancer, endometrial cancer, liver cancer, colon cancer, thyroid cancer, pancreatic cancer, colorectal cancer, esophageal cancer, skin cancer, lymphoma, and multiple myeloma; the above immune disease includes one or more of connective tissue disease, systemic sclerosis, rheumatoid arthritis, and systemic lupus erythematosus; the above cardiovascular disease includes one or more of angina pectoris, myocardial infarction, stroke, heart attack, hypertensive heart disease, rheumatic heart disease, cardiomyopathy, cardiac arrhythmia, and congenital heart disease; the above metabolic disease includes one or more of diabetes mellitus, gout, obesity, hypoglycemia, hyperglycemia, and dyslipidemia; and the above neurological disease includes one or more of Alzheimer's disease, Parkinson's disease, Huntington's disease, head injury, multiple sclerosis, vertigo, coma, and epilepsy. Preparation. Claim 26 delete Claim 27 delete Claim 28 delete Claim 29 delete Claim 30 delete Claim 31 delete

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