Co-amorphous forms of β-lactoglobulin and drug substance

Abstract

The present invention relates to a co-amorphous form of a drug substance and a protein, more particularly β-lactoglobulin, wherein the purity of the β-lactoglobulin is at least 92% (w / w) of the total amount of protein contained in the co-amorphous form. The present invention also relates to compositions, such as pharmaceutical compositions, comprising the co-amorphous form.

Description

[Technical field]

[0001] The present invention relates to a co-amorphous form of a drug substance and a protein, more preferably β-lactoglobulin. The present invention also relates to compositions comprising the co-amorphous form, such as pharmaceutical, cosmetic, veterinary, food or dietary compositions. [Background technology]

[0002] Oral delivery is the preferred method of drug administration because oral formulations are inexpensive to produce and convenient for patients. However, oral formulations of poorly water-soluble crystalline drug substances are a major challenge for the pharmaceutical industry because these substances exhibit poor solubility and low dissolution rates, resulting in low bioavailability and poor therapeutic performance.

[0003] To address these issues, amorphous formulations have been used in the past. By converting the crystalline form of a drug to its amorphous counterpart, the solubility and dissolution rate of the drug substance are increased, leading to improved bioavailability and therapeutic efficacy (Hancock et al., Pharm. Res. 17 (2000) pp.397-404). However, amorphous drug forms are physically unstable and tend to recrystallize into poorly soluble crystalline forms during storage (Laitinen et al., Int. J. Pharm. 453 (2013) pp. 65-79). Therefore, methods for stabilizing amorphous drug forms are justified by the pharmaceutical industry. In particular, there is a need in the art for new excipients that can further improve the stability and / or dissolution properties of co-amorphous formulations.

[0004] Albreht et al. (J. Agric. Food Chem., 2012, 60, 10834-10843) disclosed the increase in the solubility of shikonin using β-lactoglobulin. The purity of the β-lactoglobulin used in these experiments was 90%. Furthermore, Albreht et al. did not mention the co-amorphous form of shikonin and β-lactoglobulin.

[0005] WO 2018 / 113890 discloses co-amorphous forms of drug substances and various proteins. One of these proteins is β-lactoglobulin. However, the purity of β-lactoglobulin was not specified, and the β-lactoglobulin used in the examples was derived from bovine milk with a standard purity of about 90% (Sigma-Aldrich, Germany). Furthermore, in terms of intrinsic dissolution tests and dissolution enhancement using amorphous physical stability, the best performing proteins were found to be protein mixtures, especially whey protein isolates (WPIs) containing about 50 to about 70% β-lactoglobulin.

[0006] In WO 2018 / 113890, intrinsic dissolution was used as it minimizes the contribution of particle size effects or dispersion effects in the dissolution medium and is a frequently used dissolution evaluation technique. Without being bound to a particular theory, intrinsic dissolution provides general insight into potential performance but may not necessarily reflect the true dissolution behavior of a formulation or compound. Often, the dissolved amounts in intrinsic dissolution experiments are so small that they remain far below the saturation concentration of the drug in the dissolution medium (sink conditions). Thus, they do not provide information on the possibility of supersaturation or precipitation inhibition of the formulation. These properties can be evaluated using powder dissolutions that allow the formulation to become supersaturated (non-sink conditions).

[0007] Surprisingly, it was found that β-lactoglobulin with a higher purity performed better in terms of powder dissolution and physical stability than both WPI and β-lactoglobulin with standard purity.

[0008] β-lactoglobulin having a higher than standard purity may be prepared according to WO 2018 / 115520. Summary of the Invention

[0009] In one aspect the invention relates to a co-amorphous form of drug substance and β-lactoglobulin, wherein the purity of β-lactoglobulin is at least 92% (w / w) of the total amount of protein contained in the co-amorphous form.

[0010] In a further aspect, the present invention relates to the use of β-lactoglobulin having a purity of at least 92% in the preparation of a co-amorphous form with a drug substance.

[0011] In another aspect of the present invention, the present invention relates to a pharmaceutical composition comprising the co-amorphous form according to the present invention and at least one pharma- ceutically acceptable carrier or excipient. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0012] definition

[0013] In the context of the present invention, the term "co-amorphous" refers to a combination of two or more components that form a homogeneous amorphous system in which the components are intimately mixed at the molecular level. "Co-amorphous" samples are prepared by melt- and solvent-based approaches such as spray drying, solvent evaporation, freeze drying, precipitation from supercritical fluids, melt quenching, hot melt extrusion, electrospinning, 2D printing, 3D printing, or by kinetic disordering processes such as ball milling and cryomilling. Powder X-ray diffraction (XRPD) along with differential scanning calorimetry (DSC) are used to identify whether a sample is "co-amorphous" after preparation, for example by measuring the absence of Bragg peaks and the appearance of a single glass transition temperature.

[0014] In the context of the present invention, the term "purity" in relation to the β-lactoglobulin contained in the co-amorphous form according to the present invention is defined as a percentage (w / w) of the total amount of protein contained in the co-amorphous form. When the co-amorphous form is contained in a pharmaceutical composition, any additional protein, such as gelatin, that may be contained as an excipient in the pharmaceutical formulation is not included in the calculation of the purity of the β-lactoglobulin contained in the co-amorphous form. Furthermore, when an additional protein is contained in the pharmaceutical composition as an excipient, said additional protein may generate an additional, second glass transition temperature (if amorphous) or melting point (if crystalline) in addition to the glass transition temperature of said co-amorphous form.

[0015] In the context of the present invention, the term "drug substance" is intended to refer to an active pharmacological ingredient, a dietary supplement, or a veterinary drug. In one embodiment, the term "drug substance" refers to an active pharmacological ingredient. Reference to "a" drug substance in the context of the present invention may refer to one or more drug substances.

[0016] Co-amorphous morphology

[0017] In one aspect, the present invention relates to a co-amorphous form of a drug substance and β-lactoglobulin, the purity of β-lactoglobulin being at least 92% (w / w) of the total amount of protein contained in the co-amorphous form. Without being bound to a particular theory, it has been found that the purity of β-lactoglobulin contributes positively to a higher solubility and / or stability of the drug substance. Thus, in one embodiment of the present invention, the purity of β-lactoglobulin in the co-amorphous form of the present invention is at least 94% (w / w) of the total amount of protein contained in the co-amorphous form. In another embodiment of the present invention, the purity of β-lactoglobulin in the co-amorphous form of the present invention is at least 95% (w / w) of the total amount of protein contained in the co-amorphous form. In yet another embodiment of the present invention, the purity of β-lactoglobulin in the co-amorphous form of the present invention is at least 96% (w / w) of the total amount of protein contained in the co-amorphous form. In yet another embodiment of the invention, the purity of β-lactoglobulin in the co-amorphous form of the invention is at least 97% (w / w) of the total amount of protein contained in the co-amorphous form. In yet another embodiment of the invention, the purity of β-lactoglobulin in the co-amorphous form of the invention is at least 98% (w / w) of the total amount of protein contained in the co-amorphous form.

[0018] The co-amorphous form of the present invention may comprise 1-99% of the drug substance, for example 5-95% of the drug substance. In one embodiment, the co-amorphous form comprises 10-90% of the drug substance and 10-90% of β-lactoglobulin. In a further embodiment, the co-amorphous form comprises 20-90% of the drug substance and 10-80% of β-lactoglobulin. In yet a further embodiment, the co-amorphous form comprises 30-85% of the drug substance and 15-70% of β-lactoglobulin. In another embodiment, the co-amorphous form comprises 50-85% of the drug substance and 15-50% of β-lactoglobulin. In a further embodiment, the co-amorphous form comprises 55-75% of the drug substance and 25-45% of β-lactoglobulin. In yet another embodiment, the co-amorphous form comprises 30% of the drug substance and 70% of β-lactoglobulin. In yet a further embodiment, the co-amorphous form comprises 50% drug substance and 50% β-lactoglobulin. In yet a further embodiment, the co-amorphous form comprises 60% drug substance and 40% β-lactoglobulin. In yet another embodiment, the co-amorphous form comprises 70% drug substance and 30% β-lactoglobulin.

[0019] It has been found that lower drug loading provides particularly good dissolution of drug molecules with low solubility, especially drug molecules with very low solubility. Thus, in one embodiment, the co-amorphous form comprises 5-35% (w / w) drug substance and 65-95% (w / w) β-lactoglobulin. In a further embodiment, the co-amorphous form comprises 10-30% (w / w) drug substance and 70-90% (w / w) β-lactoglobulin. In an even further embodiment, the co-amorphous form comprises 12-25% (w / w) drug substance and 75-88% (w / w) β-lactoglobulin. In an even further embodiment, the co-amorphous form comprises 15-20% (w / w) drug substance and 80-85% (w / w) β-lactoglobulin.

[0020] In a further aspect, the present invention relates to the use of β-lactoglobulin having a purity of at least 92% (w / w) in the preparation of a co-amorphous form with a drug substance.

[0021] The co-amorphous form may be prepared by the general method disclosed in this Example or in WO 2018 / 113890.Further, in one aspect, the present invention relates to a method for preparing the co-amorphous form of the present invention, said method being selected from subjecting the drug substance and β-lactoglobulin together to spray drying, solvent evaporation, freeze drying, precipitation from supercritical fluids, melt quenching, hot melt extrusion, electrospinning, 2D printing, 3D printing, and any milling process such as ball milling and cryomilling.

[0022] drug substance

[0023] Most new pharma- ceutical active molecules are very hydrophobic and difficult to dissolve in water. Examples of such molecules are those classified in classes II and IV of the Biopharmaceutical Classification System (BCS). Thus, these pharma- ceutical active molecules typically require solubilization to improve their bioavailability in the final formulation. Thus, in one embodiment, the invention relates to a co-amorphous form of drug substance and β-lactoglobulin, the purity of which is at least 92% (w / w) of the total amount of protein contained in the co-amorphous form, and the drug substance is classified in class II or IV of the BCS. In a further embodiment, the crystalline drug substance has a solubility of less than 0.1 mg / ml in water at 25°C. In yet a further embodiment, the crystalline drug substance has a solubility of less than 0.02 mg / ml in water at 25°C.

[0024] The concept is intended to be general in nature, i.e. applicable to any kind of drug substance for which improved amorphous physical stability and / or solubility would be beneficial. The drug substance may be classified as poorly soluble or insoluble, poorly soluble or impermeable, and / or slowly soluble according to the biopharmaceutical classification system. The drug substance may be selected from the following list: abiraterone acetate, aceclofenac, acetaminophen, acetazolamide, acetylsalicylic acid, acrylinium bromide, acyclovir, afamelanotide acetate, albendazole, albuterol sulfate, aliskiren fumarate, allopurinol, alprostadil, amantadine hydrochloride, aminolevulinic acid hydrochloride, amiodarone hydrochloride, amoxicillin, amprenavir, anagrelide hydrochloride, anidulafungin, apalutamide. , apixaban, apremilast, aprepitant, apriprazole, atorvastatin, azelaic acid, azithromycin, benidipine, bazedoxifene acetate, bedaquiline fumarate, benzonatate, bexarotene, bicalutamide, binimetinib, bisacodyl, brivaracetam, budesonide, candesartan, carbamazepine, cabergoline, carfilzomib, carisoprodol, carvedilol, cefdinir, cefditoren, cefixime, cefotiam, cefporin, Doxime, cefuroxime axetil, celecoxib, clarithromycin, chloroquine, chlorpromazine, ciclesonide, cilexetil, cilostazol, ciprofloxacin, cladribine, clarithromycin, clofazimine, clonazepam, clopidogrel, clozapine, cobicistat, colisthemate sodium, cyclosporine, cyproterone, dabrafenib mesylate, dapaglifozin, dapsone, daptomycin, dasabuvir, dasatinib, deferasin ROX, delafloxacin meglumine, dexamethasone, dexmethylphenidate hydrochloride, diazepam, diclofenac, diloxanide, docetaxel, dolutegravir sodium, doxycycline, dutasteride, duvelisib, ebastine, efavirenz, eluxadrine, elvitogravir, empagliflozin, enasidenib mesylate, enzalutamide, epalrestat, eprosartan, erythromycin, eslicarbazepine acetate, estradiol,Estrone sulfate, ethyl icosapentate, etoposide, etravirine, everolimus, ezetimibe, famotidine, fenofibrate, flibanserin, fluocinonide, flurbiprofen, fluticasone furoate, fluticasone propionate, folic acid, formoterol fumarate, furosemide, gefitinib, glatiramer acetate, glibenclamide, gliclazide, glimpiride, glipizide, glycopyrrolate, griseofulvin, haloperidol, hydrochlorothiazide, hydrocortisone, hydroxyzine, ibuprofen, ibrutinib, icosapentaenoic acid Pentethyl, imatinib, indinavir, irbesartan, irinotecan, isotretinoin, itraconazole, ivacaftor, ivermectin, ketoprofen, L-carbocysteine, lamotrigine, lenalidomide, lesinurad, letermovir, levalbuterol tartrate, levodopa, levonorgestrel, linezolid, lopinavir, loratadine, lorazepam, lovastatin, lubiprostone, manidipine, mebendazole, medroxyprogesterone, mefloquine, megestrol acetate, melatonin, meloxicam, melphalan, menatetreno , mercaptopurine, mesalamivir, metaxalone, methylphenidate, metoclopramide, metoprolol, metronidazole, midostaurin, modafinil, mometasone furoate, morphine sulfate, mosapride, mycamine, nabilone, nabumetone, nalidixic acid, naproxen sodium, nelfinavir, nepafenac, nevirapine, neratinib, nicergoline, niclosamide, nifedipine, nilotinib, nilvadipine, nimesulide, nimodipine, nintedanib, nitisinone, nitrofurantoin, norrethindrone acetate, nystatin, oran Zapine, olaparib, olmesartan, omadacycline, opicapone, orlistat, ospemifene, oxcarbazepine, oxycodone, paclitaxel, paliperidone palmitate, palonosetron hydrochloride, paricalcitol, pazopanib hydrochloride, perampanel, phenobarbital, phenytoin, pioglitazone, pitavastatin, posaconazole, pranlukast, praziquantel, prednisolone acetate, prednisone, progesterone, pyrantel, pyrimethamine, quetiapine, quinine, raloxifene, rebamipide, regorafenib,Retinol, ribociclib succinate, rifampicin, rifaximin, rilpivirine, rimegepant, riociguat, risperidone, ritonavir, rivaroxaban, rofecoxib, rolapitant hydrochloride, roxithromycin, rucaparib, safinamide mesylate, saquinavir, sennoside A, sertraline, sevelamer carbonate, sildenafil, simeprevir, simvastatin, sirolimus, sofosbuvir, sonidegib phosphate, sorafenib tosylate, spironolactone, sufentanil citrate, gammadex sodium, sulfadiazine, sulfamethoxazole, sulfasalazine, sultamicillin, sulpiride, sunitinib malate, sucralose Borexant, tacrolimus, tadalafil, tafamidis, tafamidis meglumine, tamoxifen, tasimelteon, tecobirimat, telaprevir, telmisartan, telotristat ethyl, teprenone, teriflunomide, theophylline, ticlopidine, tipranavir, tocopherol nicotinate, tolterodine tartrate, topotecan hydrochloride, tosufloxacin, tretinoin, triflusal, trimethoprim, umeclidinium bromide, uridine triacetate, ursodeoxycholic acid, valproic acid, valsartan, vandetanib, vemurafenib, venetoclax, verapamil, voriconazole, warfarin, ziprasidone hydrochloride and zaltoprofen.

[0025] Some drug substances contain functional groups that are alkaline and form salts with acids. Other drug substances contain functional groups that are acidic and form salts with bases. Typically, drug substances containing one or more acidic functional groups are less likely to undergo protonation in gastric acid, thus maintaining low solubility of the drug substance in gastric acid. On the other hand, drug substances containing one or more alkaline functional groups increase the solubility of the drug substance in gastric acid due to protonation. The benefits obtained with the co-amorphous form of the present invention are not limited by the presence or absence of acidic and alkaline groups, and therefore it is envisioned that the present invention is useful for any type of drug substance.

[0026] β-Lactoglobulin

[0027] β-lactoglobulin is the major whey protein in the milk of ruminants and many other mammals. Whey refers to the liquid supernatant remaining after the caseins of milk are precipitated and removed (during cheese making). However, β-lactoglobulin may also be isolated directly from milk. Bovine β-lactoglobulin is a 162 amino acid protein with a molecular weight of about 18.4 kDa. Under physiological conditions, the protein is predominantly dimeric (open form) and dissociates to a monomeric state (closed structure) below a pH of 3. The pH is very important for the crystallization of bovine β-lactoglobulin, which may form different lattices depending on the pH. Several genetic variants of β-lactoglobulin have been identified, the main bovine β-lactoglobulins are designated A and B. In one embodiment of the present invention, the β-lactoglobulin is β-lactoglobulin obtained from a mammalian species such as cow, sheep or goat, in its native and / or glycosylated form, including genetic variants. Modifications, including additions, deletions, substitutions of amino acids in naturally occurring forms of β-lactoglobulin and variants thereof, or recombinant forms of the protein, are also contemplated as part of the present invention to be useful in the present invention. In a further embodiment, the β-lactoglobulin is bovine β-lactoglobulin.

[0028] Pharmaceutical Compositions

[0029] The co-amorphous form of the present invention may be included in a pharmaceutical composition. Thus, in one aspect, the present invention relates to a pharmaceutical composition comprising a co-amorphous form according to the present invention and at least one pharma- ceutically acceptable carrier or excipient.

[0030] The co-amorphous form of the present invention is preferably formulated with a pharmaceutically acceptable carrier or excipient. The pharmaceutically acceptable carrier or excipient is an inert carrier or excipient suitable for each administration method, and is formulated into ordinary pharmaceutical preparations (tablets, granules, capsules, powders, solutions, suspensions, emulsions, injections, drip infusions, etc.). Examples of such carriers or excipients include binders, lubricants, disintegrants, etc., which are pharmaceutically acceptable. When used as an injection solution or an infusion suspension, it is formulated using distilled water for injection, physiological saline, glucose solution, etc.

[0031] The administration method of the pharmaceutical composition of the present invention is not particularly limited, and ordinary oral or parenteral administration methods (intravenous, intramuscular, subcutaneous, transdermal, intranasal, transmucosal, enteral, etc.) are applicable. In one embodiment, the pharmaceutical composition is in a form suitable for oral administration or intranasal administration, such as solid preparations, powders, tablets, capsules, granules, sachets, reconstitutable powders, powders, dry powder inhaler drugs, and chewable agents.

[0032] It should be understood that any of the above-mentioned features and / or aspects in relation to the compounds described in the present invention are applied by analogy with the methods described herein.

[0033] The following figures and examples are provided below to illustrate the present invention. They are intended to be illustrative and should not be construed in a limiting way in any manner.

Brief Description of the Drawings

[0034] [Figure 1] Powder dissolution of crystalline compound A and co-amorphous formulations at 50% (WPI, ALA, BLG98, and cGMP) and 60% (WPI, ALA, and BLG98) drug load in 0.1M HCl and FaSSIF-V2. [Diagram 2]Powder dissolution of crystalline Compound B and co-amorphous formulations at 50% (WPI, ALA, BLG98 and cGMP), 60% (WPI, ALA, BLG98 and cGMP) and 70% (ALA and BLG98) drug loading in 0.1 M HCl and FaSSIF-V2. [Diagram 3] Powder dissolution of co-amorphous formulations at 50% (BLG98 and BLG90, respectively) Compound B drug loading in 0.1 M HCl and FaSSIF-V2. [Figure 4] Powder dissolution of co-amorphous formulations at 50% (WPI, BLG98 and BLG90, respectively) indomethacin drug loading in 0.1 M HCl and FaSSIF-V2. [Diagram 5] XRPD diffractograms of pure as-milled drug Compound A and co-amorphous formulations at 50%, 60% and 70% (w / w) drug loading in combination with proteins WPI, ALA, BLG98, cGMP. [Figure 6] XRPD diffractograms of freshly milled pure drug compound B and co-amorphous formulations at 50% and 60% (w / w) drug loading in combination with proteins WPI, ALA, BLG98, and cGMP. At 70% (w / w) drug loading, co-amorphous were prepared only with ALA and BLG98. [Figure 7] XRPD diffractograms of stored samples of Drug Compound A. The diffractograms shown indicate whether the samples remained amorphous (5 week halo) or the appearance of crystalline peaks at the time they first appeared during the stability study (indicated by week number). [Figure 8] XRPD diffractograms of stored samples of drug compound B. Pure amorphous drug showed crystallinity already after 1 week of storage, whereas all co-amorphous formulations investigated at 50%, 60% and 70% (w / w) drug loading showed an amorphous halo. [Figure 9]XRPD diffraction patterns of stored samples (40 °C / 75% RH) of the drug indomethacin in co-amorphous formulations with WPI, BLG90 and BLG98 at a drug loading of 50% each. The diffraction patterns shown indicate whether the samples remained amorphous (halo for 1 month) or, in the case of BLG90, the appearance of crystalline peaks after 1 week during the stability test. [Figure 10] XRPD diffraction patterns of stored samples (ambient conditions) of the drug indomethacin in co-amorphous formulations with WPI, BLG90 and BLG98 at a drug loading of 50% each. The diffraction patterns shown indicate whether the samples remained amorphous (halo for 1 month) or, in the case of BLG90, the appearance of crystalline peaks after 1 week during the stability test. [Figure 11] XRPD diffraction patterns of various freshly milled co-amorphous formulations with a 50% drug loading in combination with BLG98 and BLG90. [Figure 12] Powder dissolution of crystalline compounds APA, BDQ, RIF, RIT, and VNX and their respective co-amorphous formulations with a 50% drug loading in combination with BLG98 and BLG90 in 0.1 M HCl and FaSSIF-V2. [Figure 13] XRPD diffraction patterns of freshly milled and stored co-amorphous formulations containing the compound IND with a 30% drug loading in combination with BLG98 and BLG90. [Figure 14] Powder dissolution of the co-amorphous formulation of the compound IND with a 30% drug loading in combination with BLG98 and BLG90 in 0.1 M HCl and FaSSIF-V2. The co-amorphous formulation was obtained by ball milling. [Figure 15] XRPD diffraction pattern of a freshly spray-dried co-amorphous formulation with a 50% RIF drug loading in combination with BLG98 and BLG90. [Figure 16] Powder dissolution of the co-amorphous formulation with a 50% RIF drug loading in combination with BLG98 and BLG90 in 0.1 M HCl and FaSSIF-V2. The co-amorphous formulation was obtained by spray drying.

Example

[0035] material

[0036] Drug Compound A and Drug Compound B are small molecule compounds. Compound A (melting point (T m ) = 284 °C, logP = 1.8, pKa = 6.3 (acidic) and 9.8) has a solubility of 0.02 mg / ml in water at 25 °C and a solubility of 0.02 mg / ml at 25 °C and pH 1. m = 259 °C, log P = 2, neutral) has a solubility of 0.01 mg / ml in water at 25 °C and 0.3 mg / ml at pH 1 (25 °C). m = 162 °C, logP = 4.3, pKa = 4.5 (acidic)) were purchased from Hawkins, Inc. (Minneapolis, MN, USA). Whey protein isolate (WPI), β-lactoglobulin (BLG98) with purity >98% in the protein fraction, α-lactalbumin (ALA) and casein glycomacroprotein (cGMP) were obtained from Arla Food Ingredients. β-lactoglobulin (BLG90) with a purity of approximately 90% in the protein phase was obtained from Sigma-Aldrich.

[0037] Apalutamide (APA), bedaquiline fumarate (BDQ), nimodipine (NMD), rifaximin (RIF), ritonavir (RIT), and venetoclax (VNX) are small molecule active compounds with distinct physicochemical properties, including acidic, basic, and neutral molecules as well as ionic compounds in the form of salts.

[0038] method

[0039] Ball Milling

[0040] Protein-based co-amorphous forms were prepared using vibrating ball milling (MixerMill MM400, Retsch GmbH & Co., Haan, Germany) for 60 min at 30 Hz in a cold room at 4° C. For this purpose, a total mass of 500 mg of material at the respective mass ratio between protein and drug (30%, 50%, 60% or 70% drug loading) was weighed into a 25 ml milling jar and milling was performed with two 12 mm stainless steel balls.

[0041] Spray drying

[0042] Protein-based co-amorphous forms were prepared by using a Buchi B-290 spray dryer (Buchi Labortechnik AG, Falwil, Switzerland) equipped with a three-fluid nozzle (Buchi Labortechnik AG, Falwil, Switzerland), an inert loop B-295 (Buchi Labortechnik AG), and a dehumidifier (Buchi Labortechnik AG). Compound RIF was dissolved in ethanol (anhydrous, ≥ 99.8%) at a concentration of 20 mg / ml as the inner feed, and BLG98 or BLG90 was dissolved in water at a concentration of 20 mg / ml and used as the outer feed. The inner and outer feeds were pumped separately into the spray dryer at a constant feed rate of 1.8 ml / min. The spray drying process was carried out with the following process settings: inlet temperature 100°C, drying air flow rate approximately 35 m 3 / h, and atomizing air flow rate of 473 l / h. The outlet temperature was recorded at 65-70°C.

[0043] X-ray powder diffraction (XRPD) for the determination of solid morphology

[0044] The presence of fully amorphous or crystalline formulations was determined using an X'Pert PANanalytical PRO X-ray diffractmeter (PANanalytical, Almelom, The Netherlands) with Cu Kα radiation (λ = 1.54187 Å). Samples were scanned in reflectance mode from 5° to 30° 2θ with a scan speed of 0.067° 2θ / s and a step size of 0.026° 2θ. Acceleration voltage and current were 45 kV and 40 mA, respectively.

[0045] Powder dissolution test in 0.1M HCl, FaSSGF and FaSSIF

[0046] Powder dissolution of samples was determined at room temperature in either 0.1 M HCl or fasted state simulated intestinal fluid V2 (FaSSIF V2, Biorelevant) as dissolution medium. Samples equivalent to 20 mg of drug were added to a 100 ml Erlenmeyer flask containing 20 ml of dissolution medium. A magnetic stir bar was added to the Erlenmeyer flask containing the dissolution medium and stirred at 200 rpm. At pre-determined time points (5, 10, 20, 40, 60, 90, 120 min), 2 ml of dissolution medium was removed from the dissolution vessel and immediately replaced with 2 ml of fresh dissolution medium. Dissolution samples were filtered through a 0.45 μm filter, dissolved using acetonitrile, and then filtered again through a 0.45 μm filter. Finally, samples were analyzed for drug content using high performance liquid chromatography (HPLC) for Compound A, Compound B, Compound IND, Compound NMD, Compound RIF, Compound RIT, and Compound VNX (with BLG90); or UV spectroscopy for Compound APA, Compound BDQ, and Compound VNX (with BLG98). For HPLC analysis, an Agilent 1260 infinity HPLC system equipped with an Agilent 1290 Diode Array Detector (Agilent, Santa Clara, USA) was used. The column was an Agilent 5TC-C18(2) 250 *4.6 mm, 5 μm, injection volume was 20 μl. Flow rate was 1 ml / min for all compounds. Quantification of Compound A, Compound B, Compound RIF and Compound RIT used a 5TC-C18(2) (Agilent, 4.6×150 mm, 5 μm) column. Quantification of Compound VNX (with BLG90) used an Eclipse XDB-C18 (Agilent, 4.6×150 mm, 5 μm) column.

[0047] For compound A, the mobile phase consisted of 15 mM ammonium dihydrogen phosphate in water and acetonitrile in a volume ratio of 3:7, while for compound B, the mobile phase consisted of 0.05% TFA in water and acetonitrile in a volume ratio of 4:6. The UV detection wavelengths were 225 nm and 248 nm for compound A and compound B, respectively. The retention times were about 3.9 minutes and 4.3 minutes for compound A and compound B, respectively. For indomethacin, the mobile phase consisted of 1.25% phosphoric acid in water and methanol in a volume ratio of 15:85. The UV detection wavelength was 240 nm, and the retention time was about 5.5 minutes.

[0048] For compound RIF, the mobile phase was 20 volumes of 3.16 g / l ammonium formate (pH 7.2±0.05) and an equal mixture of 80 volumes of acetonitrile and methanol. The UV detection wavelength was 276 nm and the retention time was about 5.6 min. For compound RIT, the mobile phase was 2 g / l KH 2 PO 4 It was a mixture of water and acetonitrile. 3 PO 4 The mobile phase was adjusted to pH 4.0±0.05 by using a chromatograph. The UV detection wavelength was 215 nm, and the retention time was about 11.7 minutes. For compound VNX (using BLG90), the mobile phase was 10 times the amount of 25 mM ammonium formate (pH 6.5) and 90 times the amount of acetonitrile. The UV detection wavelength was 250 nm, and the retention time was about 5.3 minutes.

[0049] For compounds APA, BDQ, and VNX (with BLG98), samples were analyzed using an Evolution 300 UV spectrophotometer at 320 nm (Thermo Scientific, Cambridge, UK).

[0050] physical stability

[0051] All samples containing Compound A and Compound B were stored in a desiccator at 40° C. in saturated sodium chloride solution to obtain a relative humidity of 75% (40° C. / 75% RH). Samples containing Compound A, Compound B and IND [IND at 50% (w / w) drug loading] were examined for solid state by XRPD at day 0 and subsequently after 1, 3 and 5 weeks. Samples containing indomethacin were stored at both 40° C. / 75% RH and ambient conditions and analyzed after 1 week and 1 month of storage. Samples containing IND at 30% (w / w) drug loading were examined for solid state by XRPD at day 0 and subsequently after 3 weeks.

[0052] Tuning Temperature Differential Scanning Calorimetry (mDSC) for Determining Glass Transition Temperature (Tg) and Homogeneity of Co-Amorphous Morphologies

[0053] mDSC thermograms of the samples were collected using a Discovery DSC (TA instruments, New Castle, USA) under a nitrogen gas flow of 50 ml / min. Samples containing compound A, compound B, and IND were analyzed at a heating rate of 2°C / min from 25°C to 200°C with a base modulation temperature amplitude of 0.2120°C and a period of 40 seconds. The same heating rate, amplitude, and period were applied for the remaining samples. Samples containing compounds APA, BDQ, RIF, and VNX were heated from 25°C to 250°C, and samples containing compound RIT were heated from 0°C to 170°C. A total of 4-8 mg of sample powder was filled into an aluminum Tzero pan and sealed with an aluminum Tzero lid. The glass transition temperature (Tg) was determined as the midpoint from the reversing heat flow.

[0054] Example 1 - Powder dissolution of co-amorphous formulations (Drugs Compound A, Compound B, and Indomethacin)

[0055] For compound A loaded at 50% (w / w), the co-amorphous formulations with WPI, ALA, and BLG98 released approximately 90% of compound A and performed equally in 0.1 M HCl and FaSSIF (Figure 1). However, for compound A loaded at 60% (w / w), the co-amorphous formulations with WPI, ALA, and BLG98 performed differently. In the dissolution medium 0.1 M HCl, the co-amorphous formulations with ALA and BLG98 initially released approximately 80% of compound A (5 min to 60 min), after which the formulation with ALA showed precipitation of the drug, whereas the formulation with BLG98 maintained its concentration level and showed no sign of precipitation. The co-amorphous formulation with WPI reached a slightly lower concentration releasing approximately 75% compound A after 10 min, and then declined slightly but continuously to a concentration releasing approximately 70% compound A at 120 min. Thus, WPI performed overall less well than BLG98 (throughout the experiment) and ALA (first 60 min of the experiment). In the dissolution medium FaSSIF, the co-amorphous forms with ALA and BLG98 performed comparably, releasing about 80% Compound A and maintaining that concentration level without any signs of precipitation. The co-amorphous formulations with WPI reached slightly lower concentrations releasing about 75% Compound A, performing overall less well than ALA and BLG98 (throughout the experiment). Finally, all co-amorphous formulations (50% and 60% drug loading) performed better than pure crystalline Compound A.

[0056] For Compound B loadings of 50% and 60% (w / w), the co-amorphous formulations with WPI, ALA, BLG98, and cGMP released approximately 90% and 80% of Compound B, respectively, in 0.1 M HCl, which was retained until the end of the experiment (Figure 2). Similarly, for Compound B loadings of 70% (w / w), the co-amorphous formulations with ALA and BLG98 released approximately 80% of Compound B in 0.1 M HCl, which was retained until the end of the experiment.

[0057] In FaSSIF, compound B co-amorphous formulations with WPI, ALA, and BLG98 at 50% and 60% (w / w) loadings initially released about 70% of compound B (10 min), followed by precipitation of compound B to a concentration level of about 40% drug release. Compound B co-amorphous formulations with ALA and BLG98 at 70% (w / w) loadings performed similarly. Conversely, compound B co-amorphous formulations with cGMP at 50% and 60% (w / w) loadings performed inferior to all other formulations in FaSSIF, releasing about 20% of the total amount of compound B.

[0058] Finally, all co-amorphous formulations (50, 60% and 70% drug loading) performed better than pure crystalline Compound B.

[0059] At a loading of 50% (w / w) compound B, the co-amorphous formulations with BLG98 and BLG90 released about 85 and 80%, respectively, in 0.1 M HCl, and about 45 and 30%, respectively, in FaSSIF V2 (Figure 3). This demonstrated that β-lactoglobulin with higher purity provides improved solubility and dissolution for compound B in both acidic and neutral solvents.

[0060] For indomethacin at a loading of 50% (w / w), the co-amorphous formulation with BLG98 showed higher release than both WPI and BLG90 in 0.1 M HCl, with BLG90 outperforming WPI (Figure 4). In FaSSIF, the different protein grades showed similar final release profiles; however, BLG98 reached a plateau sooner than BLG90 (Figure 4).

[0061] Overall, considering the results of the dissolution test, the pure crystalline drug performed inferior to any co-amorphous formulation. Within the co-amorphous formulation, cGMP performed inferior to WPI, ALA, and BLG98. Considering the results of Compound A only, BLG98 was superior to ALA and WPI at 60% (w / w) drug loading and appeared equivalent at 50% (w / w) drug loading. For Compound B, similar results were obtained for WPI, ALA, and BLG98 in terms of dissolution behavior. For Compound B and indomethacin, BLG98 was clearly superior to BLG90, and for indomethacin, it was also superior to WPI. Thus, the higher purity of β-lactoglobulin provides improved properties compared to the prior art form with 90% purity.

[0062] Example 2 - Physical stability of co-amorphous samples containing drugs Compound A, Compound B, and indomethacin

[0063] XRPD was used to analyze the solid state of the samples. XRPD was used to analyze the solid state of the samples. Amorphous material is indicated by the appearance of an amorphous halo structure in the XRPD, i.e., the absence of any Bragg peaks in the diffractogram, while the presence of crystallinity was identified by the presence of crystalline peaks in the diffractogram. Figures 5 and 6 show the appearance of an amorphous halo in each case, proving successful amorphization for either the pure drug Compound A and Compound B, or all drug-protein mixtures. Physical stability was performed under humid conditions at 40°C and 75% RH in open vials.

[0064] Upon storage, the pure amorphous drugs Compound A and Compound B were found to be unstable, showing the appearance of crystalline peaks already within one week of storage (Figures 7 and 8). For Compound A with loadings of 50% and 60% (w / w), the co-amorphous formulations with ALA and BLG98 remained amorphous over the entire period (5 weeks), whereas the co-amorphous formulations with WPI or cGMP showed crystalline peaks after one week (Compound A-cGMP 60%), three weeks (Compound A-cGMP 50%, Compound A-WPI 50%) or five weeks (Compound A-WPI 60%) of storage (Figure 7). For Compound A with loadings of 70% (w / w), the co-amorphous formulations with WPI, ALA and cGMP showed crystalline peaks after one week, whereas BLG98 remained amorphous after one week and showed crystalline peaks after three weeks. For Compound B loadings of 50%, 60%, and 70% (w / w), all investigated co-amorphous formulations remained amorphous over the entire period (5 weeks).

[0065] Indomethacin stored under accelerated (40°C / 75%RH) and ambient conditions at 50% drug loading in co-amorphous formulations with WPI, BLG90 and BLG98, respectively, demonstrated improved stability compared to BLG90 (Figures 9 and 10). After one month, BLG98 still has an amorphous halo, while BLG90 shows a crystalline peak after one week. WPI also maintains the amorphous form longer than BLG90.

[0066] Overall, considering the stability study results, the pure drug required an amorphous stabilizer and comparatively BLG98 performed the best.

[0067] Example 3 - Thermal analysis of co-amorphous formulations

[0068] Table 1 reveals that the Tg of pure drug Compound A and Compound B are both low for any co-amorphous formulation. The appearance of a single Tg in any mDSC thermogram of the co-amorphous formulation suggests that all formulations, in combination with all proteins, WPI, ALA, BLG98 and cGMP, resulted in homogeneous single-phase amorphous systems. Furthermore, it can be seen that all proteins, WPI, ALA, BLG98 and cGMP result in Tgs with similar values ​​for each drug loading. For those samples that remained amorphous after 5 weeks of storage, the Tg was reanalyzed and found to remain very similar to the freshly prepared Tg, indicating that storage did not alter the homogeneity of these co-amorphous formulations. [Table 1]

[0069] Example 4 - Preparation of co-amorphous formulations at 50% drug loading in combination with BLG98 or BLG90 obtained by ball milling, diffraction analysis, thermal analysis and powder dissolution (drugs APA, BDQ, RIF, RIT, and VNX)

[0070] The as-prepared co-amorphous formulations at 50% drug loading showed the appearance of an amorphous halo (Figure 11) as well as a single glass transition temperature (Table 2), suggesting that all formulations in combination with either BLG98 or BLG90 led to homogeneous single-phase amorphous systems. [Table 2]

[0071] At a drug loading of 50% (w / w), co-amorphous formulations containing compounds APA, BDQ, RIF, RIT, and VNX with BLG98 or BLG90 showed a substantial increase in dissolution rate and solubility in both dissolution media compared to the respective pure crystalline compounds ( FIG. 12 ). Additionally, it was found that the co-amorphous formulations prepared with BLG98 generally showed faster dissolution and higher solubility compared to the respective co-amorphous formulations prepared with BLG90.

[0072] Overall, BLG98 provides improved dissolution and solubility for a variety of compounds with different physicochemical properties at a drug loading of 50% (w / w) compared to the respective crystalline drug and co-amorphous formulations prepared in BLG90.

[0073] Example 5 - Preparation of co-amorphous formulation at 30% drug loading in combination with BLG98 obtained by ball milling, physical stability, thermal analysis and powder dissolution (drug IND)

[0074] To test whether lower drug loadings could potentially improve drug release, co-amorphous formulations containing the compound IND with BLG98 or BLG90 at a drug loading of 30% (w / w) were prepared by ball milling. As shown in Figure 13, the freshly prepared co-amorphous formulations at a drug loading of 30% (w / w) showed a Tg (IND-BLG98-30%) = 141.0℃ and Tg (IND-BLG90-30%)= 144.1 °C, suggesting that both formulations in combination with BLG98 and BLG90 resulted in a single-phase amorphous system. At a loading of 30% (w / w) of compound IND, the co-amorphous formulations with BLG98 or BLG90 reached a drug release of about 800 μg / ml in both 0.1 M HCl and FaSSIF (Figure 14). For the dissolution medium 0.1 M HCl, the dissolution was faster and much higher concentrations were obtained compared to the drug release of the co-amorphous formulation of compound IND with BLG98 at a drug loading of 50% (w / w) (about 25 μg / ml, see Figure 4). For the dissolution medium FaSSIF, similar concentrations were obtained for the co-amorphous formulation of compound IND with BLG98 at a drug loading of 50% (w / w) (Figure 4). Therefore, the results suggest that drug loadings less than 50% (w / w) can increase the dissolution performance and solubility of drugs from co-amorphous formulations with BLG.

[0075] Example 6 - Powder dissolution and physical stability of co-amorphous formulations obtained by spray drying (Drug RIF)

[0076] As shown in FIG. 15, a freshly prepared spray-dried co-amorphous formulation containing RIF at a drug loading of 50% (w / w) showed a Tg (RIF-BLG98-50%) = 198.4℃ and Tg (RIF-BLG90-50%) = 199.2 °C, suggesting that the resulting spray dried formulations resulted in a single phase amorphous system in combination with BLG98 and BLG90. With regard to dissolution behavior (Figure 16), the two spray dried co-amorphous formulations showed similar drug release in the first 20 minutes, however, after 20 minutes, the co-amorphous formulation of RIF with BLG98 remained stable at the concentration of dissolved RIF, whereas the co-amorphous formulation of RIF with BLG90 showed precipitation and was unable to maintain the drug in its supersaturated state. Furthermore, higher drug release was obtained from the spray dried material compared to the dissolution obtained from the ball milled co-amorphous formulation (Figure 12).

Claims

1. 1. A co-amorphous form of an active pharmacological ingredient, a dietary supplement or a veterinary drug with β-lactoglobulin, characterized in that the purity of the β-lactoglobulin is at least 92% (w / w) of the total amount of protein contained in the co-amorphous form, and the active pharmacological ingredient, dietary supplement or veterinary drug is classified as class II or IV of the Biopharmaceutical Classification System (BCS).

2. The co-amorphous form of claim 1, wherein the active pharmacological ingredient, nutritional supplement, or veterinary drug has a solubility of less than 0.1 mg / ml in water at 25°C.

3. 3. The co-amorphous form of claim 1 or 2, wherein the purity of the β-lactoglobulin is at least 94% (w / w) of the total amount of protein contained in the co-amorphous form.

4. 4. The co-amorphous form of claim 3, wherein the purity of the β-lactoglobulin is at least 96% (w / w) of the total amount of protein contained in the co-amorphous form.

5. 5. The co-amorphous form of claim 4, wherein the purity of the β-lactoglobulin is at least 97% (w / w) of the total amount of protein contained in the co-amorphous form.

6. 6. The co-amorphous form of claim 5, wherein the purity of the β-lactoglobulin is at least 98% (w / w) of the total amount of protein contained in the co-amorphous form.

7. 7. The co-amorphous form of any one of claims 1 to 6, wherein the β-lactoglobulin is bovine β-lactoglobulin.

8. 8. The co-amorphous form of any one of claims 1 to 7, wherein the co-amorphous form comprises 5-85% (w / w) of the active pharmacological ingredient, nutraceutical, or veterinary drug and 15-95% (w / w) of β-lactoglobulin.

9. 9. The co-amorphous form of any one of claims 1 to 8, wherein the co-amorphous form comprises 5-35% (w / w) of the active pharmacological ingredient, nutraceutical, or veterinary drug and 65-95% (w / w) of β-lactoglobulin.

10. 10. The co-amorphous form of any one of claims 1 to 9, wherein the active pharmacological ingredient, dietary supplement, or veterinary drug has a solubility of less than 0.02 mg / ml in water at 25°C.

11. The co-amorphous form of any one of claims 1 to 10, wherein the active pharmacological ingredient, dietary supplement, or veterinary drug is a dietary supplement.

12. A pharmaceutical composition comprising the co-amorphous form as defined in any one of claims 1 to 11 and at least one pharma- ceutically acceptable carrier or excipient.

13. 2. Use of beta-lactoglobulin having a purity of at least 92% (w / w) for preparing a co-amorphous form with an active pharmacological ingredient, a dietary supplement, or a veterinary medicinal product, where the active pharmacological ingredient, the dietary supplement, or the veterinary medicinal product is classified as class II or IV of the Biopharmaceutical Classification System (BCS).

14. 14. The use according to claim 13, wherein the purity of the beta-lactoglobulin is at least 94% (w / w).

15. 12. A method for preparing the co-amorphous form according to any one of claims 1 to 11, the method being selected from subjecting the active pharmacological ingredient, nutraceutical or veterinary drug together with β-lactoglobulin to spray drying, solvent evaporation, freeze drying, precipitation from supercritical fluids, melt quenching, hot melt extrusion, electrospinning, 2D printing, 3D printing, and any milling process such as ball milling and cryomilling.

16. The method of claim 15, wherein the method is spray drying.

17. 17. The method according to claim 16, wherein the spray drying is carried out either by using a two-fluid nozzle, where both the active pharmacological ingredient, dietary supplement or veterinary drug and the beta-lactoglobulin are dissolved together in the same solvent or solvent mixture and subsequently subjected to a spray drying process; or by using a three-fluid nozzle, where the active pharmacological ingredient, dietary supplement or veterinary drug and the beta-lactoglobulin are dissolved in separate solvents or solvent mixtures and when spraying, the solvents are combined in a spray drying process via a three-fluid nozzle.

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