Subcutaneously administered antibiotic pharmaceutical composition

Abstract

The present invention discloses an antibiotic pharmaceutical composition for intradermal or subcutaneous administration, comprising an antibiotic and hyaluronidase. The present invention also relates to a kit comprising said pharmaceutical composition, a method for preparing said kit, and uses of said pharmaceutical composition and kit.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical preparations, and specifically, to an antibiotic pharmaceutical composition suitable for intradermal or subcutaneous administration, a kit comprising said composition and a method for preparing the same, as well as the use of said composition and kit.

Background Art

[0002] Antibiotics are the most commonly used drugs for clinical anti-infection. As a result of years of development, there are thousands of types of antibiotics, and hundreds of types are commonly used clinically. The main classifications include β-lactam, aminoglycoside, macrolide, lincomycin, polypeptide, quinolone, sulfonamide, anti-tuberculosis drugs, antifungal drugs and other antibiotics.

[0003] The administration methods of antibiotics are various, and oral administration, intramuscular administration, and intravenous administration are common. However, the administration of antibiotics by these methods has limitations, drawbacks, and side effects.

[0004] Intravenous administration is a method of directly injecting a drug into the body by drip or bolus. This method is immediate and has high bioavailability, so it is a commonly used administration method clinically, but it is also recognized as one of the most dangerous administration methods. In addition to the risk of infection due to deep intervention, it is inevitable that insoluble particles will be mixed into the drug during manufacturing, transportation, storage, and dispensing, causing side effects such as vascular embolism, phlebitis, fever, and granuloma. Also, some antibiotics are unstable in the gastrointestinal tract and are easily destroyed by the gastrointestinal tract, and some can only be administered intravenously. Furthermore, the operation of intravenous drug administration is technically difficult and usually requires professional medical staff in medical facilities, which causes inconvenience to patients' treatment and consumes a large amount of medical resources. Intravenous drug administration requires intravenous puncture, which may be difficult for some people with poor vascular conditions, such as the elderly, infants, children, and severely burned patients.

[0005] Oral administration is the most common method of drug therapy, and its advantages include convenience, painlessness, ease of administration, low cost of drug delivery, safety compared to intravenous administration and other methods, and a low incidence of injection-related side effects. Currently, despite the advantages of oral antibiotics, their disadvantages are becoming apparent. In addition to potentially irritating the gastrointestinal tract and adversely affecting the digestive system, studies have shown that oral antibiotics may affect infant development. Oral administration of antibiotics has been shown to inhibit the colonization of gut microbiota, leading to risks such as an increased incidence of asthma and type 1 diabetes. Furthermore, in cancer patients, the use of oral antibiotics in parallel with or prior to immunotherapy has been shown to disrupt the gut microbiota and significantly reduce the effectiveness of immunotherapy (Routy, B et al. (2018). Science 359(6371): 91-97).

[0006] Intramuscular injection has several drawbacks, including the inability to administer volumes greater than 5 mL, the tendency to cause pain and discomfort to patients, a slow onset of action, and the possibility of nerve damage due to improper injection. For these reasons, it has rarely been used as a method of administering antibiotics.

[0007] Intradermal or subcutaneous administration refers to injecting medication into the skin or subcutaneous tissue so that the drug is rapidly absorbed. Subcutaneous administration is less expensive than intravenous administration, offers more site options, is easier to insert, causes less pain and discomfort than intravenous insertion, is easier to re-insert the needle at a different site, and allows for insertion into non-sensitive areas of the skin. Furthermore, because intradermal or subcutaneous administration can be performed in virtually any environment, it is more appropriate than intravenous fluid administration in situations where care is limited. However, if the volume of intradermal or subcutaneous fluid exceeds 2 mL, it can cause tissue deformation, increase interstitial pressure, cause pain and discomfort, and limit the volume and rate of subcutaneous fluid administration at a single site. Due to limitations in fluid volume and the potential for skin irritation from high concentrations of antibiotics, antibiotics are almost always administered intravenously or orally in clinical practice.

[0008] Therefore, there is an urgent need to develop antibiotics that are safe, fast-acting, and can be administered subcutaneously without causing discomfort to patients. [Overview of the project] [Problems that the invention aims to solve]

[0009] In view of the shortcomings of the prior art, the object of the present invention is to provide a pharmaceutical composition for intradermal or subcutaneous administration, wherein the antibiotic pharmaceutical composition comprises an antibiotic and hyaluronidase. The pharmaceutical composition has significant advantages over oral, intramuscular, or intravenous administration of antibiotics. For example, the blood drug concentration (Cmax) of the antibiotic in the composition is increased compared to administration of the antibiotic alone, the time to maximum blood concentration (Tmax) of the antibiotic in the composition is decreased compared to administration of the antibiotic alone, and / or subcutaneous administration of the composition results in a significantly smaller decrease in the number and diversity of the gut microbiota compared to pre-administration compared to oral administration of the antibiotic. [Means for solving the problem]

[0010] Specifically, the present invention relates to the following items:

[0011] 1. A pharmaceutical composition for intradermal or subcutaneous administration, comprising an antibiotic and hyaluronidase.

[0012] 2. The pharmaceutical composition according to item 1, wherein the antibiotic content is 10 mg / mL to 5 g / mL, and the hyaluronidase content is 45 units / mL to 4,500,000 units / mL.

[0013] 3. The pharmaceutical composition according to 1 or 2 above, wherein the antibiotic is selected from the group consisting of β-lactam, aminoglycoside, macrolide, lincomycin, polypeptide, tetracycline, quinolone, sulfonamide, antituberculosis drug, and antifungal drug.

[0014] 4. The β-lactam antibiotics include penicillin G, penicillin V, fenesillin, oxacillin, methicillin, cloxacillin, dicloxacillin, ampicillin, amoxicillin, pivampicillin, carbenicillin, piperacillin, sulbenicillin, temocillin, mezlocillin, amdinocillin, pibmecillinum, aparcillin, aspoxicillin, azidocillin, azulocillin, bacampicillin, benzylpenicillin phosphate, benzylpenicillin sodium, kalindacillin, clometocillin, cyclacillin, epicillin, fenbenicillin, flucloxacillin, and Tacillin, Renanpicillin, Methampicillin, Methicillin sodium, Nafcillin, Penamecillin, Penetamate hydriozide, Penicillin G benetamine, Penicillin G benzathine, Penicillin G benzhydrylamine, Penicillin G calcium, Penicillin G hydravamin, Penicillin G potassium, Penicillin G procaine, Penicillin N, Penicillin O, Penicillin V benzathine, Penicillin V hydravamin, Penimepicycline, Phenesicillin potassium, Propicillin, Quinacillin, Sulbenicillin, Sultamicillin, Tarampicillin, Tazocillin, TicalcillinLoracarbef, 3-chloro-1-carbacephem, 3-thiosubstituted carbasephem, flomoxef, latamoxycef, moxalactam, cefazolin, cephalexin, cephalothin, cefradiol, ceffuroxime, cefaclor, cephamandol, cefotiam, cefonisid, cefolanide, cefoperazone, ceftriaxone, ceftazidime, cefotaxime, ceftizoxime, cefixime, cefozidime, cefpyramide, cefpirome, cefepime, cefclizine, cefadroxil, cefatoridine, cefazedone, cefcapene pivoxil, cefclizine, cefdinir, cefditoren, cefetamet, cefme Selected from the group consisting of noxime, cefotetan, cefozopran, cefpimisole, cefpodoxime proxetil, cefprodil, ceffloxazine, cefsulodine, cefteram, ceftibuten, ceftizoxime, cefzonam, cefacetril, cephaloglysin, cephaloridine, cephalosporin, cephalothin, cefapillin, cefubperazone, cefminox, imipenem, meropenem, panipenem, biapenem, ertapenem, faropenem, cefoxitin, cefmetazole, aztreonam, carmonam, moxalactam, flomoxef, clavulanate, clavulanate, clavulanate, sulbactam, and tazobactam. The aminoglycoside antibiotic is selected from the group consisting of streptomycin, kanamycin, gentamicin, amikacin, tobramycin, netylmycin, shisomicin, apramycin, arbekacin, bambermycin, butyrosine, dibekacin, dihydrostreptomycin, forthymicin, isepamycin, micronomycin, neomycin, neomycin undecylenate, paromomycin, ribostamycin, spectinomycin, and trospectomycin. The aforementioned macrolide antibiotics are selected from the group consisting of erythromycin, clarithromycin, azithromycin, roxithromycin, carbomycin, dylithromycin, erythromycin assistrate, erythromycin estrate, erythromycin single ceptate, erythromycin lactobionic acid, erythromycin propionate, erythromycin stearate, josamycin, leucomycin, medemycin, myocamycin, oleandmycin, primycin, rokitamycin, rosamycin, spiramycin, and troleandmycin. The lincomycin-type antibiotic is selected from the group consisting of lincomycin and clindamycin. The polypeptide antibiotics mentioned above are selected from the group consisting of polymyxin B, polymyxin E, norvancomycin, teicoplanin, vancomycin, teicoplanin, amphomycin, bacitracin, capreomycin, colistimetamethasone, colistin, enzulacidine, enviomycin, fusafundin, gramicidin, mikamicin, polymyxin, pristinamycin, dalfopristin, ristocetin, thiostrepton, tuberactinomycin, tyrosidine, tyrotricin, biomycin, virginiamycin, and bacitracin zinc. The tetracycline antibiotics are selected from the group consisting of apicycline, chlortetracycline, chromocycline, demeclocycline, doxycycline, guamecycline, lymecycline, meclocycline, metacycline, minocycline, oxytetracycline, penimepicycline, pipacycline, lolitetracycline, sancycline, tetracycline, cycloserine, mupirocin, and tuberine. The aforementioned quinolone antibiotics are selected from the group consisting of nalidixic acid, pipemidic acid, norfloxacin, ofloxacin, ciprofloxacin, enoxacin, pefloxacin, levofloxacin, gatifloxacin, moxifloxacin, norfloxacin, freloxacin, lomefloxacin, sparfloxacin, glepafloxacin, rufloxacin, clinafloxacin, varofloxacin, trovafloxacin, fluoroquinolones, allatrofloxacin mesylate, cinoxacin, difloxacin, flumequin, glepafloxacin, miloxacin, marbofloxacin, nadifloxacin, quinic acid, pazufloxacin, pyromidic acid, losoxacin, temafloxacin, tosufloxacin, and trovafloxacin mesylate. The aforementioned sulfonamide antibiotics include sulfamethoxazole, trimethoprim, acetylsulfamethoxypyrazine, benzylsulfamide, chloramine B, chloramine T, dichloramine T, sulfisomidine, β-glucosylsulfanilamide, mafenide, 4'-methylsulfamoylsulfanilamide, noprilsulfamide, phthalylsulfacetoamide, phthalylsulfathiazole, sulfasalazine, succinylsulfathiazole, sulfabenzamide, sulfacetoamide, sulfachlorpyridazine, sulfacrysodine, sulfacitin, sulfadiazine, sulfadiclamide, sulfadimethoxine, sulfadoxin, sulfaethidol, sulfaguanidine, and sulfagua Selected from the group consisting of ol, sulfarene, sulfaloxic acid, sulfamerazine, sulfameta, sulfamerazine, sulfamethizol, sulfametomidine, sulfamethoxazole, sulfamethoxypyridazine, sulfametol, sulfamide chrysoidine, sulfamoxol, sulfanilamide, 4-sulfanilamide salicylic acid, sulfanilamide, sulfanilylurea, n-sulfanilyl-3,4-xylamide, sulfanitran, sulfaperine, sulfafenazole, sulfaproxin, sulfapyrazine, sulfapyridine, sulfasomisole, sulfazimazine, sulfathiazole, sulfathiourea, sulfatramide, sulfadimethine, and sulfisoxazole, The aforementioned furan antibiotics are selected from the group consisting of nitrofurantoin, furazolidone, flartadone, furazolium chloride, nifladen, nifuratel, nifluforine, niflupyrinol, nifluprazine, and niflutoinol. The nitroimidazole antibiotic is selected from the group consisting of metronidazole, tinidazole, and ornidazole. The aforementioned anti-tuberculosis antibiotic is selected from the group consisting of isoniazid, rifampicin, rifamide, pyrazinamide, ethambutol, and / or The aforementioned antifungal antibiotics include amphotericin B, fluconazole, itraconazole and 5-flucytosine, candicidine, dermostin, filimaricin, fungichromin, trichomycin, hamycin, lusensomycin, mepaltricin, natamycin, natacin, nystatin, variotin, perimycin, azacerin, griseofulvin, oligomycin, neomycin undecylenate, pyrrolnitrin, siccinin, tubercidine, pyridine, allylamine, butenafine, naphtifin, terbinafine, imidazole, bifonazole, butoconazole, chlordantoin, chlormidazole, croconazole, clotrimazole, econazole, enilconazole, fenticonazole, flutrimazole, isoconazole, ketoconazole, lanoconazole, miconazole, omoconazole, and oxy A pharmaceutical composition as described in 3 above, selected from the group consisting of conazole nitrate, sertaconazole, sulconazole, thioconazole, voriconazole, thiocarbamate, tolcyclate, trindate, tolnaftate, triazole, saperconazole, terconazole, acrisolcin, amorolphin, bifenamine, bromosalicylcloranilide, buclossamide, calcium propionate, chlorphenesin, copalafinate, diamtasol dihydrochloride, exalamide, flucytosine, haretasol, hexetidine, lipopeptides such as echinocandin, roflucarban, nifuratel, potassium iodide, propionic acid, 2-mercaptopyridine-n-oxide, salicylanilide, sodium propionate, sulfentine, tenonitrozole, triacetin, ujothion, undecylenic acid, and zinc propionate.

[0015] 5. The pharmaceutical composition according to any one of items 1 to 4 above, wherein the hyaluronidase has hyaluronic acid degrading activity under neutral conditions, and preferably the hyaluronidase is selected from the group consisting of animal testicular extract hyaluronidase, recombinant animal hyaluronidase or its variants, recombinant and / or extracted bacterial hyaluronidase, recombinant human hyaluronidase or its variants, preferably the hyaluronidase is recombinant human hyaluronidase or its variants, more preferably the hyaluronidase is recombinant human hyaluronidase or its variants, and more preferably the recombinant human hyaluronidase consists of the amino acid sequence shown in SEQ ID NO. 1 in the sequence listing, and most preferably the amino acid sequence of the recombinant human hyaluronidase is shown in SEQ ID NO. 2 in the sequence listing.

[0016] 6. The pharmaceutical composition according to any one of items 1 to 5 above, wherein the enzymatic activity of the hyaluronidase is 45 units / ml to 3,000,000 units / ml, preferably 45 units / ml to 1,500,000 units / ml, more preferably 50 units / ml to 30,000 units / ml, and most preferably 100 units / ml to 3,000 units / ml.

[0017] 7. The pharmaceutical composition according to any one of claims 1 to 6, wherein the Cmax of the antibiotic in the composition is increased by at least 10%, for example, at least 20%, at least 30%, at least 40%, and at least 50%, compared to administration of the antibiotic alone, and / or the Tmax of the antibiotic in the composition is decreased by at least 20%, for example, at least 30%, at least 40%, and at least 50%, compared to administration of the antibiotic alone, and / or the AUClast of the antibiotic in the composition is increased by at least 5%, and at least 10%, compared to administration of the antibiotic alone.

[0018] 8. The pharmaceutical composition according to any one of items 1 to 7 above, wherein, after subcutaneous administration of the composition, the amount of reduction in the diversity of the intestinal flora compared to before administration is 90% or less of the amount of reduction after oral administration of an antibiotic compared to before administration, for example, 80% or less, 70% or less, 60% or less, or 50% or less.

[0019] 9. The antibiotic pharmaceutical composition according to any one of items 1 to 8 above, wherein the pharmaceutical composition optionally contains pharmaceutically acceptable additives.

[0020] 10. The pharmaceutical composition according to 9 above, wherein the pharmaceutically acceptable additive is selected from the group consisting of buffers, stabilizers, nonionic surfactants, cosolvents, preservatives and / or excipients.

[0021] 11. The buffer solution is selected from the group consisting of histidine buffer, acetate buffer, phosphate buffer, citrate buffer, and Tris buffer, preferably a phosphate buffer, and most preferably disodium hydrogen phosphate. The aforementioned stabilizer is selected from the group consisting of trehalose, sucrose, mannitol, sodium chloride, methionine, and disodium edetate, preferably trehalose, mannitol, methionine, and sucrose, and most preferably a combination of methionine and trehalose. The nonionic surfactant is selected from the group consisting of polysorbate 20, polysorbate 80, and poloxamer 188, preferably polysorbate 20 and polysorbate 80, and most preferably polysorbate 20. The cosolvent is selected from the group consisting of sodium carbonate, phosphoric acid, citric acid, sodium bicarbonate, sodium hydroxide, sodium chloride, and L-arginine, and is preferably sodium bicarbonate and sodium hydroxide. The aforementioned preservative is selected from the group consisting of glycerol, methylparaben, propylparaben, benzoic acid, sodium benzoate, and / or, The excipient is selected from the group consisting of sorbitol, mannitol, trehalose, glycerol, lactose, sucrose, trehalose, maltose, and glucose, preferably mannitol, trehalose, sucrose, and most preferably a combination of trehalose and mannitol, the pharmaceutical composition according to 10 above.

[0022] 12. The concentration of the buffer is 1 to 100 mM, preferably 5 to 50 mM, such as 5 mM, 10 mM or 50 mM, The concentration of the stabilizer is 1 to 500 mM, preferably 30 to 150 mM, such as 5 mM methionine + 25 mM trehalose, 5 mM methionine + 53 mM trehalose, 5 mM methionine + 100 mM trehalose, 10 mM methionine + 25 mM trehalose, 10 mM methionine + 53 mM trehalose, 10 mM methionine + 100 mM trehalose, 50 mM methionine + 25 mM trehalose, 50 mM methionine + 53 mM trehalose, 50 mM methionine + 100 mM trehalose, The concentration of the excipient is 1 to 500 mM, preferably 160 to 280 mM, such as 160 mM, 220 mM or 280 mM, The concentration of the surfactant is 0.01 to 0.1% (w / v), preferably 0.02 to 0.04% (w / v), most preferably 0.02% (w / v), and / or The concentration of the co-solvent is 0.01 g / L to 100 g / L, the pharmaceutical composition according to 11 above.

[0023] 13. A lyophilized preparation and / or a liquid preparation, the pharmaceutical composition according to any one of 1 to 12 above.

[0024] 14. A kit comprising the pharmaceutical composition according to 1 to 13 above, wherein the antibiotic and the hyaluronidase kit are packaged mixed or separately.

[0025] 15. The kit according to 14, wherein the separately packaged antibiotic and / or separately packaged hyaluronidase are lyophilized or liquid formulations.

[0026] 16. The kit according to 15, wherein the lyophilized formulation is reconstituted before being administered to a subject, and the liquid formulation is administered directly to a subject or diluted before being administered to a subject.

[0027] 17. The kit according to any one of items 14 to 16, wherein the antibiotic and the hyaluronidase are administered sequentially or simultaneously, preferably the separately packaged antibiotic and / or separately packaged hyaluronidase are administered sequentially or simultaneously.

[0028] 18. The kit according to 17, wherein the sequential administration of the antibiotic and the hyaluronidase is performed by a three-way infusion tube, and the administration rate is preferably controlled by manual pushing, an infusion pump, or gravity.

[0029] 19. A method for preparing a kit as described in any one of items 15 to 18 above, (a) A step of providing an antibiotic, (b) A step of providing hyaluronidase, (c) The step of optionally providing a pharmaceutically acceptable additive, A kit preparation method comprising preparing the antibiotic and the hyaluronidase separately into lyophilized or liquid formulations, or mixing the antibiotic and the hyaluronidase to prepare a lyophilized or liquid formulation.

[0030] 20. An injection system comprising a delivery device selected from the group consisting of a syringe, an infusion pump, an injection pen, a needleless device, and the injection system being filled with a pharmaceutical composition described in any one of items 1 to 14 above.

[0031] 21. Uses of the pharmaceutical composition described in any one of items 1 to 14 above, or the kit described in any one of items 15 to 18 above, in the preparation of drugs used for the treatment of diseases selected from the group consisting of bacterial, fungal, actinomycete, mycoplasma, chlamydia, spirochete, amoeba infections and diseases resulting from said infections.

[0032] 22. The bacteria mentioned above are selected from the group consisting of the genera Escherichia, Pseudomonas, Klebsiella, Acinetobacter, Enterobacter, Citrobacter, Haemophilus, Proteus, Salmonella, Shigella, Serratia, Corynebacterium, Staphylococcus, Streptococcus, Enterococcus, Neisseria, Mycobacterium, and Legionella. The aforementioned fungi are selected from the group consisting of the genera Trichophyton, Epidermis, Microsporum, Candida, Cryptococcus, Coccidioides, Histoplasma, Sporotrichus, Blastomyces, Giotrichum, Aspergillus, Mucor, and Penicillium, for the uses described in 21 above.

[0033] 23. The bacteria or fungi mentioned above include Haemophilus influenzae type B, Pseudomonas aeruginosa types A and B, Staphylococcus aureus, Group B Streptococcus, Streptococcus pneumoniae types (1, 3, 4, 6, 7, 8, 9, 12, 14, 18, 19 and 23), Staphylococcus epidermidis, Staphylococcus saprophyticus, Streptococcus pyogenes, Streptococcus viridans, Streptococcus agalactiae, Streptococcus pneumoniae, Bacillus anthrax, Neisseria diphtheriae, Bordetella pertussis, and Tetanus. Uses as described in 21 or 22 above, selected from the group consisting of bacteria, Mycobacterium tuberculosis, Bacillus, Clostridium perfringens, Mycobacterium leprae, Neisseria gonorrhoeae, Neisseria meningitidis, Legionella, Shigella, Pseudomonas aeruginosa, Proteus, Vibrio parahaemolyticus, Escherichia coli, Salmonella typhi, Salmonella paratiphi, Salmonella, Haemophilus influenzae, Acinetobacter, Aspergillus, Candida, Cryptococcus, Mucor or Fusarium.

[0034] 24. The diseases mentioned above are chronic osteomyelitis caused by Staphylococcus aureus, endocarditis caused by drug-resistant Staphylococcus aureus or enterococcus, typhoid fever, paratyphoid fever, bacterial food poisoning, bacterial infectious diarrhea, cholera, bacterial dysentery, brucellosis, plague, anthrax, diphtheria, pertussis, scarlet fever, epidemiological spinal corditis, tuberculosis, bacterial bloodstream infection, bacterial upper respiratory tract infection, bacterial lower respiratory tract infection, bacterial urinary tract infection, bacterial abdominal infection, dermatophytosis, fungal stomatitis, candidal vaginitis, fungal pneumonia, fungal urinary tract infection, fungal bacteremia, cryptococcosis, candidiasis, aspergillosis, and Pneumocystis, as described in any one of items 21 to 23 above. [Modes for carrying out the invention]

[0035] This invention provides an antibiotic that can be administered subcutaneously. The subcutaneous antibiotic of this invention has no dosage limitations when administered, has a rapid onset of action, high blood concentration, high patient comfort, and does not cause microbiological damage to the gastrointestinal tract.

[0036] 1.Definition Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those generally understood by those skilled in the art to which this invention pertains.

[0037] "In combination with..." means administering two or more therapeutic agents to the target patient as a mixture simultaneously or sequentially as individual agents in any order.

[0038] "Pharmaceutical composition" means a product obtained by mixing an antibiotic and hyaluronidase, including fixed and unfixed combinations. Pharmaceutical compositions typically contain pharmaceutically acceptable additives. "Fixed combination" means a single pharmaceutical composition containing an antibiotic and hyaluronidase that is administered simultaneously. "Unfixed combination" means the administration of separate pharmaceutical compositions or unit dosage forms of the antibiotic and hyaluronidase as separate entities simultaneously, in parallel, or sequentially, at non-specific time intervals, thereby providing effective levels of both compounds into the subject's body.

[0039] "Antibiotics" refers to a broad class of anti-infective drugs used clinically that inhibit and destroy pathogenic microorganisms such as bacteria and fungi. As a result of years of development, thousands of antibiotics exist, with hundreds commonly used clinically, including beta-lactams, aminoglycosides, macrolides, lincomycins, polypeptides, quinolones, sulfonamides, anti-tuberculosis drugs, antifungal drugs, and other antibiotics.

[0040] Hyaluronic acid (HA), also known as hyaluronic acid or hyaluronan, is a linear high-molecular-weight glycosaminoglycan composed of repeated links of two disaccharide units: D-glucuronic acid and N-acetylglucosamine. It is widely present in the connective tissue, mucous tissue, lens of the eye, and skin of vertebrates, and is particularly abundant in tissues such as embryos, cartilage, synovial fluid, vitreous humor, umbilical cord, and comb. Hyaluronic acid is one of the most widely distributed acidic mucopolysaccharides in the human tissue matrix, forming a network barrier between the collagen fibrous skeleton of the extracellular matrix and filling the body. This hyaluronic acid barrier is also one of the most important factors in preventing the rapid absorption of drugs injected subcutaneously.

[0041] Hyaluronidase refers to an endoglycosidase capable of breaking down hyaluronic acid and hydrolyzing the β-1,4 glycosidic bonds of the HA chain to produce low molecular weight glycosidases, which in turn produce low molecular weight HA or oligosaccharides. This increases tissue permeability and improves the permeability of fluids within the tissue. Hyaluronidase has been used in the medical field for many years as a "drug dispersant," promoting the diffusion of locally stored drugs, exudate, or blood, facilitating drug absorption, reducing local tissue tension and pain, and promoting the absorption and dissipation of edema and inflammatory exudate. Clinically, hyaluronidase can also be used as a drug penetration agent, an anesthetic adjuvant, and a postoperative edema reducer.

[0042] "Activity" means functional activity or the activity of a polypeptide or a part thereof in relation to a full-length (intact) protein. Functional activity includes, but is not limited to, biological activity, catalytic or enzymatic activity, antigenicity (ability to bind to or compete with an anti-polypeptide antibody), immunogenicity, ability to form polymers, and specific binding of polypeptides to receptors or ligands.

[0043] "Recombination" primarily refers to recombinant proteins. Recombinant proteins are produced by obtaining proteins using recombinant DNA or recombinant RNA technology. The acquisition routes are divided into in vitro and in vitro methods. In both methods, the prerequisite is to obtain a recombinant vector to which a gene fragment translatable to the target protein is attached using recombinant technology, and then introduce this vector into a host cell capable of expressing the target protein to express a specific recombinant protein molecule.

[0044] A "mutant" refers to a protein whose amino acid sequence has been altered through artificial modification, thereby changing its function or properties.

[0045] "Hyaluronidase activity" refers to the ability of hyaluronidase to cleave hyaluronic acid. In vitro measurement methods for determining the hyaluronidase activity of hyaluronidase (e.g., recombinant human hyaluronidase PH20) are known in the art and are described herein. An example measurement is a trace turbidity measurement, which indirectly determines the cleavage of hyaluronic acid by hyaluronidase by detecting the formation of an insoluble precipitate when uncleaved hyaluronic acid binds to serum albumin.

[0046] "Tmax" refers to the time it takes for the blood concentration of a drug to reach its peak after a single dose.

[0047] "Cmax" refers to the maximum blood concentration value on the blood concentration-time curve, i.e., the highest plasma drug concentration that can be reached after drug administration. Peak concentration is closely related to the clinical application of a drug. The peak concentration is the effective concentration required to exert the drug's effect, and exceeding the safety margin can lead to toxic reactions. Furthermore, peak concentration is an important indicator of the absorption and safety of the drug formulation.

[0048] "Intestinal microbiota" refers to the totality of all microorganisms that inhabit the digestive tract, including bacteria, fungi, and viruses. Of the 29 known bacterial phyla, the Firmicutes and Bacteroidetes phyla play important roles in the intestinal tract of healthy individuals, followed by the Proteobacteria and Actinomycetes phyla. Intestinal microbiota are mutually restrictive and dependent, forming a symbiotic ecological balance with the host, and playing crucial roles in maintaining the integrity of the intestinal mucosa, metabolism, immune regulation, and nutritional supply. The gut microbiota is closely related to human health, and research has shown that imbalances in the gut microbiota are associated with the development and onset of diseases such as inflammatory bowel disease, colorectal cancer, obesity, and asthma.

[0049] "Gut microbiota diversity" refers to the number of species present in the gut of an animal. "Gut microbiota population" refers to the number of microorganisms, such as various classes of bacteria, present in the gut of a sample. Techniques commonly used to study gut microbiota diversity and quantity include metagenomic sequencing, 16S rRNA sequencing, metatranscriptomics, chip technology, and fluorescence quantitative PCR. Metagenomic technology can detect the genomes of all microorganisms, including bacteria, fungi, and viruses, and is one of the most comprehensive detection methods. 16S rRNA sequencing technology can detect bacteria at the genus level, assess gut microbiota diversity, provide a comprehensive indicator of the relative content of gut microbiota, and allow for prediction of gene metabolic networks using software. Metatranscriptomics technology can quantitatively detect all types of gut microbiota (bacteria, viruses, phages, archaea, fungi, yeasts, parasites) down to the "species" level and even the "strains" level. Chip technology allows for the qualitative detection of bacteria, fungi, and viruses.

[0050] "Pharmacologically acceptable additives" refer to components of a pharmaceutical composition other than the active ingredient that are not toxic to the subject. Pharmaceutically acceptable additives include, but are not limited to, buffers, excipients, stabilizers, preservatives, pH adjusters, nonionic surfactants, and / or cosolvents.

[0051] Oral administration is the most commonly used mode of drug delivery for pharmacotherapy, offering advantages such as convenience, painlessness, ease of administration, low cost, and safety compared to, for example, intravenous administration, with fewer injection-related side effects. The disadvantage of oral drug delivery is that after oral administration, the drug is absorbed slowly and irregularly from the gastrointestinal tract, resulting in generally low drug utilization. Certain drugs are prone to causing gastrointestinal problems and should not be administered orally. In addition, oral administration of certain drugs can irritate the gastrointestinal tract or adversely affect the digestive system.

[0052] Subcutaneous administration refers to the method of injecting medication into the subcutaneous tissue for absorption. Subcutaneous drug administration offers many advantages in terms of cost-effectiveness and ease of use, including: 1. Lower cost. 2. A wider range of sites are available, the needle insertion is simpler, causing less pain and discomfort than intravenous injection, making it easier to change sites and re-insert the needle, and allowing for insertion into relatively insensitive areas of the skin. 3. In situations with limited care, subcutaneous fluid administration is more appropriate than intravenous fluid administration. From a safety perspective, the presence of a structurally fixed fibrous skeleton in the subcutaneous tissue restricts the flow of molecules larger than 200 nm in diameter through the subcutaneous tissue, avoiding the risk of large insoluble particles being introduced into the central circulation. Subcutaneous fluid administration offers the following safety advantages: 1. No risk of thrombus formation. 2. Less likely to cause pulmonary edema or fluid overload. 3. No risk of thrombophlebitis. 4. No risk of complications such as sepsis or systemic infection.

[0053] "Approximately" means within an acceptable margin of error for a particular value, as determined by those skilled in the art, and this margin of error depends in part on how the value is measured or determined, i.e., on the limits of the measuring system. In the context of a particular measurement, result, or embodiment, unless expressly stated in the example or elsewhere in this specification, "approximately" means within one standard deviation or up to 5%, whichever is greater, as practiced in the art.

[0054] 2. Pharmaceutical composition of the present invention A first aspect of the present invention provides an antibiotic pharmaceutical composition for subcutaneous administration, the antibiotic pharmaceutical composition comprising an antibiotic and hyaluronidase. The content of the antibiotic is 10 mg / mL to 5 g / mL, and the concentration of the hyaluronidase is 45 units / ml to 4,500,000 units / ml, preferably 45 units / ml to 3,000,000 units / mL, more preferably 45 units / ml to 1,500,000 units / ml, more preferably 50 units / ml to 30,000 units / ml, and most preferably 100 units / ml to 3,000 units / ml.

[0055] The aforementioned antibiotic pharmaceutical composition comprises one or more antibiotics and / or recombinant human hyaluronidase, and pharmaceutically acceptable additive components.

[0056] The aforementioned antibiotics include β-lactams, aminoglycosides, macrolides, lincomycins, polypeptides, tetracyclines, quinolones, sulfonamides, antituberculosis drugs, and antifungal drugs. The β-lactam antibiotic is preferably penicillin G, penicillin V, fenesillin, oxacillin, methicillin, cloxacillin, dicloxacillin, ampicillin, amoxicillin, pivampicillin, carbenicillin, piperacillin, sulbenicillin, temocillin, mezlocillin, amdinocillin, pibmecillinum, aparcillin, aspoxicillin, azidocillin, azlocillin, bacampicillin, benzylpenicillin phosphate, benzylpenicillin sodium, kalindacillin, clomethicillin, cyclacillin, epicillin, fenbenicillin, or flucloxacillin. Hetacillin, Renanpicillin, Methampicillin, Methicillin sodium, Nafcillin, Penamecillin, Penetamate hydrozide, Penicillin G benetamine, Penicillin G benzathine, Penicillin G benzhydrylamine, Penicillin G calcium, Penicillin G hydravamin, Penicillin G potassium, Penicillin G procaine, Penicillin N, Penicillin O, Penicillin V benzathine, Penicillin V hydravamin, Penimepicycline, Phenesicillin potassium, Propicillin, Quinacillin, Sulbenicillin, Sultamicillin, Tarampicillin, Tazocillin, TicalcillinLoracarbef, 3-chloro-1-carbacefem, 3-thiosubstituted carbasefem, flomoxef, latamoxicef, moxalactam, cefazolin, cephalexin, cephalothin, cefradiol, cefuroxime, cefaclor, cephamandol, cefotiam, cefonisid, cefolanide, cefoperazone, ceftriaxone, ceftazidime, cefotaxime, ceftizoxime, cefixime, cefozidime, cefpyramide, cefpirome, cefepime, cefclizine, cefadroxil, cefatolidin, cefazedone, cefcapene pivoxil, cefclizine, cefdinir, cefditoren, cefetamet These include cefmenoxime, cefotetan, cefozopran, cefpimisole, cefpodoxime proxetil, cefprodil, ceffloxazine, cefsurodin, cefteram, ceftibuten, ceftizoxime, cefzonam, cefacetril, cephaloglysin, cephaloridine, cephalosporin, cephalothin, cefapyrine, cefbuperazone, cefminox, imipenem, meropenem, panipenem, biapenem, ertapenem, faropenem, cefoxitin, cefmetazole, aztreonam, carmonam, moxalactam, flomoxef, clavulanic acid, clavulanate, clavulanate, sulbactam, and tazobactam. The aminoglycoside antibiotic is preferably streptomycin, kanamycin, gentamicin, amikacin, tobramycin, netylmycin, shisomicin, apramycin, arbekacin, bambermycin, butyrosine, dibekacin, dihydrostreptomycin, forthymicin, isepamycin, micronomycin, neomycin, neomycin undecylenate, paromomycin, ribostamycin, spectinomycin, or trospectomycin. The macrolide antibiotics are preferably erythromycin, clarithromycin, azithromycin, roxithromycin, carbomycin, dylithromycin, erythromycin assistrate, erythromycin estrate, erythromycin single ceptate, erythromycin lactobionic acid, erythromycin propionate, erythromycin stearate, josamycin, leucomycin, medemycin, myocamycin, oleandmycin, primycin, rokitamycin, rosamycin, spiramycin, and troleandmycin. The lincomycin-based antibiotic is preferably lincomycin or clindamycin. The polypeptide antibiotics mentioned above are selected from the group consisting of polymyxin B, polymyxin E, norvancomycin, teicoplanin, vancomycin, teicoplanin, amphomycin, bacitracin, capreomycin, colistimetamethasone, colistin, enzulacidine, enviomycin, fusafundin, gramicidin, mikamicin, polymyxin, pristinamycin, dalfopristin, ristocetin, thiostrepton, tuberactinomycin, tyrosidine, tyrotricin, biomycin, virginiamycin, and bacitracin zinc. The tetracycline antibiotics are preferably apicycline, chlortetracycline, chromocycline, demeclocycline, doxycycline, guamecycline, lymecycline, meclocycline, metacycline, minocycline, oxytetracycline, penimepicycline, pipacycline, lolitetracycline, sancycline, tetracycline, cycloserine, mupirocin, and tuberine. The quinolone antibiotics are preferably nalidixic acid, pipemidic acid, norfloxacin, ofloxacin, ciprofloxacin, enoxacin, pefloxacin, levofloxacin, gatifloxacin, moxifloxacin, norfloxacin, freloxacin, lomefloxacin, sparfloxacin, glepafloxacin, rufloxacin, clinafloxacin, varofloxacin, trovafloxacin, fluoroquinolone, allatrofloxacin mesylate, cinoxacin, difloxacin, flumequin, glepafloxacin, miloxacin, marbofloxacin, nadifloxacin, quinic acid, pazufloxacin, pyromidic acid, losoxacin, temafloxacin, tosufloxacin, and trovafloxacin mesylate. The sulfonamide antibiotics are preferably sulfamethoxazole, trimethoprim, acetylsulfamethoxypyrazine, benzylsulfamide, chloramine B, chloramine T, dichloramine T, sulfisomidine, β-glucosylsulfanilamide, mafenide, 4'-methylsulfamoylsulfanilamide, noprilsulfamide, phthalylsulfacetoamide, phthalylsulfathiazole, sulfasalazine, succinylsulfathiazole, sulfabenzamide, sulfacetoamide, sulfachlorpyridazine, sulfacrysodine, sulfacitin, sulfadiazine, sulfadiclamide, sulfadimethoxine, sulfadoxin, sulfaetidol, and sulfaguanidine These include sulfaguanol, sulfarene, sulfaloxic acid, sulfamerazine, sulfameta, sulfamerazine, sulfamethizol, sulfametomidine, sulfamethoxazole, sulfamethoxypyridazine, sulfametol, sulfamide chrysoidine, sulfamoxol, sulfanilamide, 4-sulfanilamide salicylic acid, sulfanilamide, sulfanillylurea, n-sulfanillyl-3,4-xylamide, sulfanitran, sulfaperine, sulfafenazole, sulfaproxin, sulfapyrazine, sulfapyridine, sulfasomisole, sulfazimazine, sulfathiazole, sulfathiourea, sulfatramide, sulfadimethine, and sulfisoxazole. The furan antibiotic is preferably nitrofurantoin, furazolidone, flartadone, furazolium chloride, nifladen, nifuratel, nifluforine, niflupyrinol, nifluprazine, or niflutoinol. The nitroimidazole class antibiotic is preferably metronidazole, tinidazole, ornidazole. The aforementioned anti-tuberculosis antibiotic is preferably isoniazid, rifampicin, rifamide, pyrazinamide, or ethambutol. The aforementioned antifungal antibiotics are preferably amphotericin B, fluconazole, itraconazole and 5-flucytosine, candicidine, dermostin, filimaricin, fungiclomin, trichomycin, hamycin, lusensomycin, mepaltricin, natamycin, natacin, nystatin, variotin, perimycin, azacerin, griseofulvin, oligomycin, neomycin undecylenate, pyrrolnitrin, siccinin, tubercidine, pyridine, allylamine, butenafine, naphtifin, terbinafine, imidazole, bifonazole, butoconazole, chlordantoin, chlormidazole, croconazole, clotrimazole, econazole, enilconazole, fenticonazole, flutrimazole, isoconazole, ketoconazole, lanoconazole, miconazole These include omoconazole, oxiconazole nitrate, sertaconazole, sulconazole, thioconazole, voriconazole, thiocarbamate, tolcyclate, trindate, tolnaftate, triazole, saperconazole, terconazole, acrisolcin, amorolphin, bifenamine, bromosalicylcloranilide, buclossamide, calcium propionate, chlorphenesin, copalafinate, diamtasol dihydrochloride, exalamide, flucytosine, haretasol, hexetidine, lipopeptides such as echinocandin, roflucarban, nifuratel, potassium iodide, propionic acid, 2-mercaptopyridine-n-oxide, salicylanilide, sodium propionate, sulfentine, tenonitrozole, triacetin, ujothion, undecylenic acid, and zinc propionate.

[0057] The hyaluronidase described above has hyaluronic acid degrading activity under neutral conditions of pH 5.0 to pH 8.0.

[0058] Hyaluronidase is an enzyme that breaks down hyaluronic acid, reducing the viscosity of hyaluronic acid (hyaluronan) in the extracellular matrix and thereby increasing tissue permeability. Hyaluronidase is selected from hyaluronidase extracted from animal testes, recombinant animal hyaluronidase or its variants, recombinant and / or extracted bacterial hyaluronidase, recombinant human hyaluronidase or its variants, preferably recombinant human hyaluronidase or its variants, more preferably recombinant human hyaluronidase or its variants, wherein the recombinant human hyaluronidase comprises the amino acid sequence shown in SEQ ID NO. 1 in the sequence listing, and most preferably the amino acid sequence of the recombinant human hyaluronidase is shown in SEQ ID NO. 2 in the sequence listing.

[0059] The hyaluronidase, recombinant human hyaluronidase, or its variants can be used in the present invention, and these variants are described in CN1942588B, CN102943067B, CN102307993B, CN103205407B, CN104244968B, CN111971387A, and so on.

[0060] The enzymatic activity of hyaluronidase is defined in units / mL (U / mL) or total enzymatic activity (U) in a particular formulation, and is described in more detail below. The standard definition of one unit of enzymatic activity (U) is the amount of enzyme per unit time that catalyzes a limited amount of substrate reaction, e.g., 1 mole or 1 nanomolar substrate / min. Techniques for measuring the activity of hyaluronidase formulations are known in this field, and the activity of hyaluronidase formulations is usually expressed in units of U or units (hereinafter referred to as "units").

[0061] Hyaluronidase activity refers to the ability of an enzyme to break down hyaluronic acid. The method for measuring hyaluronidase provided in the Chinese Pharmacopoeia indirectly determines hyaluronidase activity by reacting the enzyme with hyaluronic acid at 37°C for 30 minutes and then measuring the amount of remaining high molecular weight hyaluronic acid substrate. During the measurement, the relative activity (in units) of any given hyaluronidase is determined using a reference standard solution. In vitro measurements for hyaluronidase are known in this field. As an exemplary measurement method, the trace turbidity measurement described below indirectly measures the cleavage of hyaluronic acid by hyaluronidase by detecting the insoluble precipitate formed when uncleaved hyaluronic acid binds to serum albumin. The reference standard can be used, for example, to create a standard curve and to determine the activity of the hyaluronidase being measured in units.

[0062] The aforementioned antibiotic pharmaceutical composition comprises one or more antibiotics, recombinant human hyaluronidase, and pharmaceutically acceptable additive components.

[0063] The combination of antibiotics and recombinant human hyaluronidase can be in the form of a combined formulation or as a separate, uncombined kit.

[0064] The Cmax of the antibiotic in the composition is increased by at least 10%, for example, at least 20%, at least 30%, at least 40%, and at least 50%, compared to administration of the antibiotic alone, and / or the Tmax of the antibiotic in the composition is decreased by at least 20%, for example, at least 30%, at least 40%, and at least 50%, compared to administration of the antibiotic alone.

[0065] After subcutaneous administration of the composition, the decrease in the diversity of the intestinal flora compared to before administration is 90% or less of the decrease after oral administration of antibiotics compared to before administration, for example, 80% or less, 70% or less, 60% or less, or 50% or less.

[0066] The composition further comprises a pharmaceutically acceptable additive, the pharmaceutically acceptable additive being selected from the group consisting of buffers, stabilizers, pH adjusters, nonionic surfactants, cosolvents, preservatives and / or excipients. The buffer is selected from the group consisting of histidine buffer, acetate buffer, phosphate buffer, citrate buffer and Tris buffer; the stabilizer is selected from the group consisting of trehalose, sucrose, mannitol, sodium chloride, methionine and disodium edetate; the pH adjuster is selected from the group consisting of phosphoric acid, acetic acid, hydrochloric acid, citric acid, sodium hydroxide and Tris; and the nonionic surfactant is selected from the group consisting of polysorbate 20, polysorbate 80 and poloxamer 188. The cosolvent is selected from the group consisting of propylene glycol, ethanol, polyethylene glycol, glycerol, sodium dodecyl sulfate, and cyclodextrin; the preservative is selected from the group consisting of glycerol, methylparaben, propylparaben, benzoic acid, sodium benzoate, and ethanol; and / or the excipient is selected from the group consisting of sorbitol, mannitol, trehalose, glycerol, lactose, sucrose, trehalose, maltose, and glucose.

[0067] The concentration of the buffer solution is 1 to 100 mM, for example 5 mM, preferably 10 mM or 50 mM, and / or The concentration of the nonionic surfactant is 0.01 to 0.5% (w / v), preferably 0.02% (w / v), and / or The concentration of the cosolvent is 0.01 g / L to 100 g / L, and / or The concentration of the aforementioned preservative is 0.1 g / L to 200 g / L, and / or The concentration of the excipient is between 1 g / L and 200 g / L.

[0068] The combination of the aforementioned drugs may be in the form of a lyophilized formulation and / or a liquid formulation, and preferably the combination of the antibiotic and recombinant human hyaluronidase may be a compound formulation or a non-compound formulation packaged separately.

[0069] 3. Kit and method of preparing the kit A second aspect of the present invention provides a kit comprising the pharmaceutical composition described above, wherein the antibiotic and the hyaluronidase kit are packaged together or separately.

[0070] The separately packaged antibiotic and / or separately packaged hyaluronidase are either lyophilized or liquid formulations. The lyophilized formulation is reconstituted before administration to a subject, and the liquid formulation is either administered directly to a subject or diluted before administration to a subject.

[0071] The antibiotic and the hyaluronidase are administered sequentially or simultaneously, preferably the separately packaged antibiotic and / or separately packaged hyaluronidase are administered sequentially or simultaneously.

[0072] The antibiotic and the hyaluronidase are administered sequentially, respectively, through a three-way infusion tube, and the administration rate is preferably controlled by manual pushing, an infusion pump, or gravity.

[0073] In the first embodiment, the kit consists of two single-dose containers, the first container being filled with the liquid formulation containing a single specific dose of recombinant human hyaluronidase in a dose of 45 units / ml to 4,500,000 units / ml, and the second container being filled with a single fixed dose of the therapeutic agent.

[0074] The container containing the recombinant human hyaluronidase liquid formulation is selected from a tubular bolt and a pre-filled needle, and is preferably a pre-filled needle.

[0075] Preferably, the recombinant human hyaluronidase liquid formulation contains 45 units / ml to 4,500,000 units / ml of recombinant human hyaluronidase.

[0076] The volume of the recombinant human hyaluronidase liquid formulation is preferably 0.1 to 50 ml, for example, 0.2 ml, 0.5 ml, 1 ml, 1.5 ml, 2 ml, 2.50 ml, 5.00 ml, 10.00 ml, 15.00 ml, 20 ml, 30 ml, 40 ml, or 50 ml.

[0077] In some embodiments, the recombinant human hyaluronidase liquid formulation contains 45 to 75,000 units of recombinant human hyaluronidase per 1 ml.

[0078] The first container is a tubular bolt, the second container is a tubular bolt, and the third container is a tubular bolt or a pre-filled needle.

[0079] The first container is a tubular bolt or a pre-filled needle, preferably a pre-filled needle, and the second container is a tubular bolt.

[0080] The recombinant human hyaluronidase liquid formulation preferably contains 45 units / ml to 100,000 units / ml of recombinant human hyaluronidase, and more preferably contains 45 units / ml to 50,000 units / ml of recombinant human hyaluronidase.

[0081] A third aspect of the present invention provides a method for preparing the above kit, the method being: (a) A step of providing an antibiotic, (b) A step of providing hyaluronidase, (c) The step of optionally providing a pharmaceutically acceptable additive, The antibiotic and the hyaluronidase are each prepared as lyophilized or liquid preparations, or the antibiotic and the hyaluronidase are mixed and prepared as lyophilized or liquid preparations.

[0082] A fourth aspect of the present invention provides a method for treating a disease using the kit of the second aspect, which is administered by separate administration or mixed administration.

[0083] The aforementioned separate administrations are, (a) The step of intradermally or subcutaneously administering the recombinant human hyaluronidase liquid formulation from the kit to the target of administration, (b) The next step is to administer the therapeutic agent in the kit to the subject.

[0084] Here, steps (a) and (b) may be performed separately, simultaneously, or alternately. If steps (a) and (b) are performed separately, the time interval between steps (a) and (b) is 0 to 24 hours. Preferably, there is no time interval, and the intervals are a maximum of 1 minute, 2 minutes, 3 minutes, 4 minutes, 5 minutes, 6 minutes, 7 minutes, 8 minutes, 9 minutes, 10 minutes, 15 minutes, 20 minutes, 25 minutes, 30 minutes, 1 hour, 2 hours, 3 hours, 6 hours, 12 hours, or 24 hours.

[0085] In the case of sequential subcutaneous administration, the time interval between steps (a) and (b) may be 0 minutes, 1 minute, 2 minutes, 3 minutes, 4 minutes, 5 minutes, 6 minutes, 7 minutes, 8 minutes, 9 minutes, 10 minutes, 15 minutes, 20 minutes, 25 minutes, 30 minutes, 35 minutes, 40 minutes, 45 minutes, 50 minutes, 55 minutes, or 60 minutes.

[0086] The aforementioned separate administrations can be achieved through three methods, resulting in simultaneous or sequential administration.

[0087] The aforementioned separate administration can be controlled by an infusion pump or gravity to control the administration rate. The recombinant human hyaluronidase liquid formulation can be administered at a rate of 0.1 to 2 ml / min. The therapeutic agent is injected at a rate of 5 ml / hour, 10 ml / hour, 30 ml / hour, 60 ml / hour, 120 ml / hour, 240 ml / hour, or 300 ml / hour.

[0088] The aforementioned mixed administration involves subcutaneously administering a mixture of the recombinant human hyaluronidase liquid preparation and the therapeutic agent from the kit to the subject, wherein the liquid preparation contains 45 to 500,000 units of recombinant human hyaluronidase per 1 ml.

[0089] The aforementioned mixed administration can be controlled by an infusion pump or gravity to control the administration rate. After the recombinant human hyaluronidase liquid formulation and the therapeutic agent are mixed, they are injected at a rate of 5 to 300 ml / hour, for example, 5 ml / hour, 10 ml / hour, 30 ml / hour, 60 ml / hour, 120 ml / hour, 240 ml / hour, or 300 ml / hour.

[0090] The aforementioned therapeutic agent for mixing is in the form of a liquid or a dry powder.

[0091] The recombinant human hyaluronidase liquid formulation is administered directly and / or diluted.

[0092] The aforementioned dilution preparation may be physiological saline, which is commonly used in this field. The aforementioned dilution ratio may be 1:10, which is commonly used in this field.

[0093] 4. Pharmaceutical applications and injection systems The present invention provides a pharmaceutical composition comprising an antibiotic and hyaluronidase, and a kit for the preparation of drugs for the treatment of diseases. In specific examples, the disease is selected from the group consisting of infections of bacteria, fungi, actinomycetes, mycoplasma, chlamydia, spirochetes, amoebas, and diseases resulting from such infections.

[0094] Preferably, the disease is caused by a bacterial or fungal infection. The bacteria include the genera Escherichia, Pseudomonas, Klebsiella, Acinetobacter, Enterobacter, Citrobacter, Haemophilus, Proteus, Salmonella, Shigella, Serratia, Corynebacterium, Staphylococcus, Streptococcus, Enterococcus, Neisseria, Mycobacterium, and Legionella. The fungi include the genera Trichomycetes, Epidermis, Microsporum, Candida, Cryptococcus, Coccidioides, Histoplasma, Sporotrichus, Blastomyces, Giotrichum, Aspergillus, Mucor, and Penicillium.

[0095] Preferably, the disease is caused by a bacterial or fungal infection, and the bacteria or fungi include Haemophilus influenzae type B, Pseudomonas aeruginosa types A and B, Staphylococcus aureus, Group B Streptococcus, Streptococcus pneumoniae types (1, 3, 4, 6, 7, 8, 9, 12, 14, 18, 19 and 23), Staphylococcus epidermidis, Staphylococcus saprophyticus, Streptococcus pyogenes, Streptococcus viridans, and Streptococcus agalactieae. It may be cocci, Streptococcus pneumoniae, Bacillus anthrax, Neisseria diphtheriae, Bordetella pertussis, Neisseria tetanus, Mycobacterium tuberculosis, Bacillus, Clostridium perfringens, Mycobacterium leprae, Neisseria gonorrhoeae, Neisseria meningitidis, Legionella, Shigella, Pseudomonas aeruginosa, Proteus, Vibrio parahaemolyticus, Escherichia coli, Salmonella typhi, Salmonella paratiphi, Salmonella, Haemophilus influenzae, Acinetobacter, Aspergillus, Candida, Cryptococcus, Mucor or Fusarium.

[0096] Diseases caused by the aforementioned pathogenic microorganisms include, for example, chronic osteomyelitis caused by Staphylococcus aureus, endocarditis caused by drug-resistant Staphylococcus aureus or enterococci, typhoid fever, paratyphoid fever, bacterial food poisoning, bacterial diarrhea, cholera, bacterial dysentery, brucellosis, plague, anthrax, diphtheria, pertussis, scarlet fever, epidemiological spinal corditis, tuberculosis, bacterial bloodstream infections, bacterial upper respiratory tract infections, bacterial lower respiratory tract infections, bacterial urinary tract infections, bacterial abdominal infections, dermatophytosis, fungal stomatitis, candidal vaginitis, fungal pneumonia, fungal urinary tract infections, fungal bacteremia, cryptococcosis, candidiasis, aspergillosis, and Pneumocystis.

[0097] Uses of the pharmaceutical composition of any one of the above preparations in the preparation of a disease-causing drug.

[0098] The present invention further relates to containers and injection systems, wherein packaging materials include, but are not limited to, tubular bolts, syringes, or test tubes, and injection systems include, but are not limited to, syringes, infusion pumps, injection pens, needleless devices, or subcutaneous patch delivery devices.

[0099] The components of the injection device include those commonly used in this art, such as a container, seal, and injection needle.

[0100] The containers include, but are not limited to, tubular bolts, syringes, or test tubes.

[0101] The material of the container may include, for example, glass or plastic, which are commonly used in this art.

[0102] The seal includes, but is not limited to, a sealing plug or a sealing ring.

[0103] The material of the seal may include, for example, rubber, plastic, or polymer materials, which are commonly used in this art.

[0104] The aforementioned injection needles include, but are not limited to, aqueous injection needles, single needles, and microneedle sets.

[0105] The material of the aforementioned injection needle is one of those commonly used in this field, such as metal, silicon, silica, glass, nickel, titanium, or biodegradable polymers.

[0106] The aqueous injection needle includes, but is not limited to, vial aqueous injection needles, ampoule aqueous injection needles, or pre-filled injection systems.

[0107] The aforementioned ampoule aqueous injection needle may be a glass ampoule or a plastic ampoule.

[0108] The aforementioned pre-filled injection system is a common one in this field, such as a pre-filled syringe.

[0109] Preferably, the container is a vial, the material is neutral borosilicate glass, and the specifications are 0.1 to 20 mL. The seal is a sealing plug, and the material is halogenated butyl rubber. The aforementioned injection needle is a single needle, a microneedle set.

[0110] Preferably, the material of the single needle is 304 or 316 stainless steel, with specifications of 30G, 24G, 27G, and 29G, and the material of the microneedle set is 304 or 316 stainless steel, a biodegradable polymer, with specifications of nano-sized needles with a height of 10 to 2000 μm and a width of 10 to 50 μm.

[0111] One embodiment of the present invention provides a product comprising the pharmaceutical composition described in the first aspect of the present invention, and a specification of use thereof. The product includes the container. The product may further include other materials expected from a commercial and user perspective, including other buffers, diluents, filters, needles, syringes, infusion pumps, and package inserts printed with instructions for use.

[0112] The above preferred conditions, which are in line with common practice in this industry, can be arbitrarily combined to obtain preferred embodiments of the present invention.

[0113] All reagents and raw materials used in this invention are commercially available.

[0114] This invention has the following positive and progressive effects, and solves the problem of subcutaneous administration of antibiotics. The bioavailability of antibiotics can reach more than 50%. Clinical use is not limited by volume, is more convenient, improves the patient's treatment experience, and can reduce fluid-related side effects. Patient comfort is high, the effect is rapid, blood concentration is high, and it does not cause gastrointestinal microbial damage.

[0115] <Examples> Example 1: Preparation of recombinant human hyaluronidase CHO cells stably expressing recombinant human hyaluronidase protein were used in suspension culture in a proprietary serum-free medium, followed by controlled culture with fluid replacement in a proprietary serum-free replacement medium, and gradually expanded to a 30L reactor scale by shake flask culture.

[0116] At 3-4 days after the start of culture, the amount of supplement medium added to the bioreactor daily was 2%-5% of the actual culture volume in the bioreactor. The culture temperature was controlled to 35°C-37°C, pH was controlled to 7.0 by adding 10% Na2CO3 and CO2, reactor aeration was controlled to 0.015-0.15 vvm, rotation speed was controlled to 80-150 rpm, and dissolved oxygen level was controlled to 20%-40%. During cell culture, samples were taken daily, and temperature, pH, glucose concentration, lactate concentration, molar permeability, and protein expression levels were monitored. When the CHO cell viability fell below 80% or the culture cycle reached 14-20 days, the culture was terminated, and recombinant human hyaluronidase culture supernatant was obtained.

[0117] The obtained recombinant human hyaluronidase supernatant was subjected to sequential deep filtration, anion chromatography, affinity chromatography, hydrophobic chromatography, cation chromatography, and ultrafiltration liquid exchange. Three batches were performed in this process, yielding recombinant human hyaluronidase stock solution with an SEC purity exceeding 95% and hyaluronidase with an RP purity exceeding 85%, with enzyme activity greater than 70,000 units / mg in both cases.

[0118] Example 2. Compatibility study of hyaluronidase and meropenem solution Solutions containing hyaluronidase and antibiotics were prepared, and the compatibility of the two substances was investigated. Dried meropenem powder for injection was dissolved in physiological saline at concentrations of 5 mg / ml and 50 mg / ml, and hyaluronidase was added to achieve final concentrations of 150, 500, 2000, and 6000 units / ml, respectively. After preparation, the solutions were left at room temperature for 15 minutes, and the appearance of the solutions was observed to measure the hyaluronidase concentration.

[0119] The study revealed that after preparing eight solutions, the solutions were clear and transparent, and no precipitate was observed. Hyaluronidase activity was measured in the solutions, and none of the meropenem solutions inhibited hyaluronidase activity. Further research is possible on the preparation of mixed hyaluronidase and meropenem solutions.

[0120] [Table 1]

[0121] Example 3. Compatibility study of hyaluronidase and ceftriaxone solution Solutions containing hyaluronidase and antibiotics were prepared, and the compatibility of the two substances was investigated. Dry ceftriaxone sodium for injection was dissolved in physiological saline at concentrations of 50 mg / ml and 250 mg / ml. Hyaluronidase was then added to achieve final concentrations of 150, 500, 2000, and 6000 units / ml, respectively. After preparation, the solutions were left at room temperature for 15 minutes, and the appearance of the solutions was observed to measure the hyaluronidase concentration.

[0122] The study revealed that after preparing eight solutions, the solutions were clear and transparent, and no precipitate was observed. Measurement of hyaluronidase activity in the solutions showed that ceftriaxone did not inhibit hyaluronidase activity. Further research can be conducted on the preparation of mixed hyaluronidase and ceftriaxone solutions.

[0123] [Table 2]

[0124] Example 4. Compatibility study of hyaluronidase and azithromycin solution The compatibility of two substances in a solution containing hyaluronidase and an antibiotic was investigated. Dry azithromycin powder for injection was dissolved in physiological saline at concentrations of 10 mg / ml and 100 mg / ml. Hyaluronidase was then added to achieve final concentrations of 150, 500, 2000, and 6000 units / ml, respectively. After preparation, the solutions were left at room temperature for 15 minutes, and the appearance of the solutions was observed to measure the hyaluronidase concentration.

[0125] The study revealed that after preparing eight solutions, the solutions were clear and transparent, and no precipitate was observed. Measurement of hyaluronidase activity in the solutions showed that azithromycin did not inhibit hyaluronidase activity. Further research can be conducted on the preparation of mixed hyaluronidase and azithromycin.

[0126] [Table 3]

[0127] Example 5. Compatibility study of hyaluronidase and cefradin solution The compatibility of two substances in a solution containing hyaluronidase and an antibiotic was investigated. Injectable cefradin dry powder was dissolved in physiological saline at concentrations of 50 mg / ml and 250 mg / ml. Hyaluronidase was then added to achieve final concentrations of 150, 500, 2000, and 6000 units / ml, respectively. After preparation, the solutions were left at room temperature for 15 minutes, and the appearance of the solutions was observed to measure the hyaluronidase concentration.

[0128] The study results showed that after preparing eight solutions, the solutions were clear and transparent, and no precipitate was observed. Measurement of hyaluronidase activity in the solutions revealed that cefradin did not inhibit hyaluronidase activity. Further research can be conducted on the preparation of mixed hyaluronidase and cefradin solutions.

[0129] [Table 4]

[0130] Example 6. Compatibility study of hyaluronidase and teicoplanin solution The compatibility of two substances in a solution containing hyaluronidase and an antibiotic was investigated. Teicoplanin dry powder for injection was dissolved in physiological saline at concentrations of 10 mg / ml and 70 mg / ml, and added to achieve final hyaluronidase concentrations of 150, 500, 2000, and 6000 units / ml, respectively. After preparation, the solutions were left at room temperature for 15 minutes, and the appearance of the solutions was observed to measure the hyaluronidase concentration.

[0131] The study results showed that after preparing eight solutions, the solutions were clear and transparent, and no precipitate was observed. Measurement of hyaluronidase activity in the solutions revealed that teicoplanin did not inhibit hyaluronidase activity. Further research can be conducted on the mixed preparation of hyaluronidase and teicoplanin.

[0132] [Table 5]

[0133] Example 7. Compatibility study of hyaluronidase and ceftazidime solution The compatibility of two substances in a solution containing hyaluronidase and an antibiotic was investigated. Injectable ceftazidime dry powder was dissolved in physiological saline at concentrations of 20 mg / ml and 200 mg / ml. Hyaluronidase was then added to achieve final concentrations of 150, 500, 2000, and 6000 units / ml, respectively. After preparation, the solutions were left at room temperature for 15 minutes, and the appearance of the solutions was observed to measure the hyaluronidase concentration.

[0134] The study results showed that after preparing eight solutions, the solutions were clear and transparent, and no precipitate was observed. Measurement of hyaluronidase activity in the solutions revealed that ceftazidime did not inhibit hyaluronidase activity. Further research can be conducted on the preparation of mixed hyaluronidase and ceftazidime.

[0135] [Table 6]

[0136] Example 8. Compatibility study of hyaluronidase and ertapenem solution The compatibility of two substances in a solution containing hyaluronidase and an antibiotic was investigated. Ertapenem dry powder for injection was dissolved in physiological saline at concentrations of 30 mg / ml and 300 mg / ml. Hyaluronidase was then added to achieve final concentrations of 150, 500, 2000, and 6000 units / ml, respectively. After preparation, the solutions were left at room temperature for 15 minutes, and the appearance of the solutions was observed to measure the hyaluronidase concentration.

[0137] The study results showed that after preparing eight solutions, the solutions were clear and transparent, and no precipitate was observed. Measurement of hyaluronidase activity in the solutions revealed that ertapenem did not inhibit hyaluronidase activity. Further research can be conducted on the preparation of mixed hyaluronidase and ertapenem solutions.

[0138] [Table 7]

[0139] Example 9. Compatibility study of hyaluronidase and polymyxin solutions The compatibility of two substances in a solution containing hyaluronidase and an antibiotic was investigated. Injectable polymyxin B dry powder was dissolved in physiological saline at concentrations of 50,000 units / ml and 250,000 units / ml. Hyaluronidase was then added to achieve final concentrations of 150, 500, 2,000, and 6,000 units / ml, respectively. After preparation, the solutions were left at room temperature for 15 minutes, and the appearance of the solutions was observed to measure the hyaluronidase concentration.

[0140] The study results showed that after preparing eight solutions, the solutions were clear and transparent, and no precipitate was observed. Measurement of hyaluronidase activity in the solutions revealed that polymyxin B did not inhibit hyaluronidase activity. Further research can be conducted on the mixed preparation of hyaluronidase and polymyxin B.

[0141] [Table 8]

[0142] Example 10. Study on Hyaluronidase Dried Powder Formulation The recombinant human hyaluronidase stock solution obtained in Example 1 was replaced with liquid formulations of different compositions, and the concentration of recombinant human hyaluronidase was adjusted to the desired concentration. All formulations were sterilized and filtered through a 0.22 μm low-protein adsorption filter, packed into sterile 5 ml glass tubular bolts under sterile conditions, sealed with fluoropolymer-laminated butyl rubber plugs, and capped with aluminum / plastic flip-off seals. The packed volume was 2 ml. These formulations were stored under different temperature conditions, and samples were taken at specified time intervals to examine formulation stability, and the stability data for different formulations were compiled. Accelerated experiments were performed by releasing the samples at 25°C, and the amount of protein was analyzed by sampling at different time intervals using the following analytical methods. The analytical methods included SEC purity, RP purity, and enzyme activity, and the experimental results are shown in Tables 2 to 6.

[0143] As shown in Table 9, recombinant human hyaluronidase under a 2 mM methionine concentration condition showed a significant decrease in RP purity and enzyme activity after being left at 25°C for a certain period of time, while recombinant human hyaluronidase preparations containing methionine in concentrations within the ranges of 5, 10, and 50 mM exhibited good stability.

[0144] As shown in Table 10, recombinant human hyaluronidase under conditions containing 250 mM trehalose and 250 mM sucrose stabilizers showed a significant decrease in SEC purity and enzyme activity after being left at 25°C for a certain period of time. The stability of recombinant human hyaluronidase preparations under stabilizer conditions containing trehalose at concentrations of 25, 53, and 200 mM, and sucrose at concentrations of 25, 53, and 200 mM, was relatively good.

[0145] As shown in Table 11, the stability of recombinant human hyaluronidase preparations under surfactant conditions containing polysorbate 20 at concentrations of 0.01-0.1%, polysorbate 80 at concentrations of 0.01-0.1%, and poloxamer 188 at concentrations of 0.01-0.1% is relatively good.

[0146] As shown in Table 12, the stability of recombinant human hyaluronidase preparations under conditions containing 160 mM to 280 mM mannitol as an excipient is relatively good.

[0147] As shown in Table 13, the recombinant human hyaluronidase activity in the recombinant human hyaluronidase liquid formulation of this example showed relatively good stability within 150, 500, 1000, 5000, 50000, 300000, 1500000, 3000000 and 4500000 units / ml, and the enzyme activity was retained at 95% or more after being left at 2-8°C for 12 months.

[0148] [Table 9]

[0149] [Table 10] [Table 11] [Table 12] [Table 13] Example 11. Hyaluronidase dried powder formulation Based on the formulation screening of Example 10, seven representative hyaluronidase formulations were selected, as shown in Table 14. For each formulation, hyaluronidase was designed at four concentrations: 150, 500, 2000, and 6000 units / ml. After lyophilization according to the lyophilization procedure shown in Table 15, the formulations were used for mixing and preparation with meropenem.

[0150] [Table 14] [Table 15] Example 12. Subcutaneous meropenem dry powder formulation A subcutaneous meropenem powder formulation was prepared by mixing freeze-dried hyaluronidase powder with meropenem powder for injection. The meropenem powder for injection contained sodium carbonate as a cosolvent. The mixture was formulated so that each portion contained 1 g of meropenem active ingredient, with hyaluronidase units of 3,000, 10,000, 40,000, and 120,000 units, respectively.

[0151] After mixing, the dried powder was taken, and 1 g of dried meropenem active ingredient powder was dissolved in 20 ml of sterile water for injection. After dissolution, the hyaluronidase concentrations corresponded to 150 units / ml, 500 units / ml, 2000 units / ml, and 6000 units / ml, respectively. The solutions were left to stand at 25°C for 15 min and 1 hr. The hyaluronidase activity, meropenem content, and appearance of the samples were measured at each sampling point. Here, both hyaluronidase activity and meropenem content were detected according to the methods of the Chinese Pharmacopoeia.

[0152] Tables 16-17 show the meropenem content (labeled composition of 90-110% is acceptable) and the changes in hyaluronidase activity under different conditions for each group.

[0153] As a result, after reconstitution, all of the different formulation solutions were clear and transparent. After 15 minutes, the meropenem content and hyaluronidase activity were stable in the different formulations. After 1 hour, the meropenem content was within the range of 90-110% in the different formulations. Hyaluronidase activity decreased slightly in groups A and B, while hyaluronidase activity was relatively well maintained in groups C, D, E, F, and G.

[0154] [Table 16] [Table 17] Example 13. Subcutaneous ceftriaxone dry powder formulation A subcutaneous ceftriaxone powder formulation was prepared by mixing freeze-dried hyaluronidase powder (formulations A-G in Table 14) with ceftriaxone powder. The mixing specifications were such that each portion contained 1 g of ceftriaxone active ingredient, and the hyaluronidase content was 600 units, 2000 units, 8000 units, and 24000 units, respectively, depending on the combination.

[0155] After mixing, the dried powder was taken, and 1 g of the dried powder containing the active ingredient ceftriaxone was dissolved in 4 ml of sterile water for injection. After dissolution, the hyaluronidase concentrations corresponded to 150 units / ml, 500 units / ml, 2000 units / ml, and 6000 units / ml, respectively. The solutions were left to stand at 25°C for 15 min and 1 hr. The hyaluronidase activity, ceftriaxone content, and appearance of the samples were measured at each sampling point. Here, both the hyaluronidase activity and ceftriaxone content were detected according to the methods of the Chinese Pharmacopoeia.

[0156] Tables 18-19 show the changes in ceftriaxone content (labeled composition of 90-110% is acceptable) and hyaluronidase activity for each group under different conditions.

[0157] As a result, after reconstitution, all of the different formulation solutions were clear and transparent. After 15 minutes, the ceftriaxone content and hyaluronidase activity were stable in the different formulations. After 1 hour, the ceftriaxone content was within the range of 90-110% in the different formulations. Hyaluronidase activity in groups A and B decreased slightly, while hyaluronidase activity in groups C, D, E, F, and G was maintained relatively well.

[0158] [Table 18] [Table 19] Example 14. Subcutaneous azithromycin dry powder formulation A subcutaneous azithromycin powder formulation was prepared by mixing freeze-dried hyaluronidase powder (formulations A-G in Table 14) with injectable azithromycin powder. Here, the injectable azithromycin powder contains anhydrous citric acid, sodium hydroxide, or phosphoric acid, with sodium hydroxide used as a cosolvent. The mixing specifications are as follows: each portion contains 1 g of azithromycin active ingredient, and the hyaluronidase content is 750 units, 2500 units, 10000 units, and 30000 units, respectively, depending on the set.

[0159] After mixing, the dried powder was taken and dissolved in 5 ml of sterile water for injection, with 0.5 g of azithromycin active ingredient in the dried powder. After dissolution, the hyaluronidase concentrations corresponded to 150 units / ml, 500 units / ml, 2000 units / ml, and 6000 units / ml, respectively. The solutions were left to stand at 25°C for 15 min and 1 hr. The hyaluronidase activity, azithromycin content, and appearance of the samples were measured at each sampling point. Here, both hyaluronidase activity and azithromycin content were detected according to the methods of the Chinese Pharmacopoeia.

[0160] Tables 20-21 show the changes in azithromycin content (labeled composition of 90-110% is acceptable) and hyaluronidase activity under different conditions for each group.

[0161] As a result, after reconstitution, all of the different formulation solutions were clear and transparent. After 15 minutes, the azithromycin content and hyaluronidase activity were stable in the different formulations. After 1 hour, the azithromycin content was within the range of 90-110% in the different formulations. Hyaluronidase activity in groups A and B decreased slightly, while hyaluronidase activity in groups C, D, E, F, and G was maintained relatively well.

[0162] [Table 20] [Table 21] Example 15. Subcutaneous cefradin dry powder formulation A subcutaneous cefradiol powder formulation was prepared by mixing freeze-dried hyaluronidase powder (formulations A-G in Table 14) with injectable cefradiol powder. Here, the injectable cefradiol powder contains arginine as a cosolvent. The mixing specifications are as follows: each portion contains 1 g of the active ingredient cefradiol, and the hyaluronidase content is 600 units, 2000 units, 8000 units, and 24000 units, respectively, depending on the combination.

[0163] After mixing, the dried powder was taken, and 1 g of the dried powder containing cefradin active ingredient was dissolved in 4 ml of sterile water for injection. After dissolution, the hyaluronidase concentrations corresponded to 150 units / ml, 500 units / ml, 2000 units / ml, and 6000 units / ml, respectively. The solutions were left to stand at 25°C for 15 min and 1 hr. The hyaluronidase activity, cefradin content, and appearance of the samples were measured at each sampling point. Here, both hyaluronidase activity and cefradin content were detected according to the methods of the Chinese Pharmacopoeia.

[0164] Tables 22-23 show the changes in cefradin content (labeled composition of 90-110% is acceptable) and hyaluronidase activity under different conditions for each group.

[0165] As a result, after reconstitution, all of the different formulation solutions were clear and transparent. After 15 minutes, the cefradin content and hyaluronidase activity were stable in the different formulations. After 1 hour, the cefradin content was within the range of 90-110% in the different formulations. Hyaluronidase activity in groups A and B decreased slightly, while hyaluronidase activity in groups C, D, E, F, and G was maintained relatively well.

[0166] [Table 22] [Table 23] Example 16. Subcutaneous teicoplanin dried powder formulation A subcutaneous teicoplanin powder formulation was prepared by mixing freeze-dried hyaluronidase powder (formulations A-G in Table 14) with teicoplanin powder. The mixing specifications were as follows: each portion contained 1 g of the active ingredient teicoplanin, and the hyaluronidase content was 750 units, 2500 units, 10000 units, and 30000 units, respectively, depending on the combination.

[0167] After mixing, the dried powder was taken and dissolved in 6 ml of sterile water for injection, with 0.4 g of teicoplanin active ingredient in the dried powder. After dissolution, the hyaluronidase concentrations corresponded to 150 units / ml, 500 units / ml, 2000 units / ml, and 6000 units / ml, respectively. The solutions were left to stand at 25°C for 15 min and 1 hr. The hyaluronidase activity, teicoplanin content, and appearance of the samples were measured at each sampling point. Here, both hyaluronidase activity and teicoplanin content were detected according to the methods of the Chinese Pharmacopoeia.

[0168] Tables 24-25 show the changes in teicoplanin content (labeled composition of 90-110% is acceptable) and hyaluronidase activity under different conditions for each group.

[0169] As a result, after reconstitution, all of the different formulation solutions were clear and transparent. After 15 minutes, the teicoplanin content and hyaluronidase activity were stable in the different formulations. After 1 hour, the teicoplanin content was within the range of 90-110% in the different formulations. Hyaluronidase activity in groups A and B decreased slightly, while hyaluronidase activity in groups C, D, E, F, and G was maintained relatively well.

[0170] [Table 24] [Table 25] Example 17. Subcutaneous ceftazidime dry powder formulation A subcutaneous ceftazidime powder formulation was prepared by mixing freeze-dried hyaluronidase powder (formulations A-G in Table 14) with ceftazidime powder. The mixing specifications were as follows: each portion contained 1 g of ceftazidime active ingredient, and the hyaluronidase content was 750 units, 2500 units, 10000 units, and 30000 units, respectively, depending on the combination.

[0171] After mixing, the dried powder was taken, and 1 g of the dried powder containing ceftazidime active ingredient was dissolved in 5 ml of sterile water for injection. After dissolution, the hyaluronidase concentrations corresponded to 150 units / ml, 500 units / ml, 2000 units / ml, and 6000 units / ml, respectively. The solutions were left to stand at 25°C for 15 min and 1 hr. The hyaluronidase activity, ceftazidime content, and appearance of the samples were measured at each sampling point. Here, both hyaluronidase activity and ceftazidime content were detected according to the methods of the Chinese Pharmacopoeia.

[0172] Tables 26-27 show the changes in ceftazidime content (labeled composition of 90-110% is acceptable) and hyaluronidase activity under different conditions for each group.

[0173] As a result, after reconstitution, all of the different formulation solutions were clear and transparent. After 15 minutes, the ceftazidime content and hyaluronidase activity were stable in the different formulations. After 1 hour, the ceftazidime content was within the range of 90-110% in the different formulations. Hyaluronidase activity in groups A and B decreased slightly, while hyaluronidase activity in groups C, D, E, F, and G was maintained relatively well.

[0174] [Table 26] [Table 27] Example 18. Subcutaneous ertapenem dry powder formulation A subcutaneous ertapenem powder formulation was prepared by mixing freeze-dried hyaluronidase powder (formulations A-G in Table 14) with ertapenem powder. The mixing specifications were such that each portion contained 1 g of the active ingredient ertapenem, and the hyaluronidase units were 500, 1667, 6667, and 20000 units, respectively, depending on the combination.

[0175] After mixing, the dried powder was taken, and 1 g of dried ertapenem active ingredient powder was dissolved in 3.2 ml of sterile water for injection. After dissolution, the hyaluronidase concentrations corresponded to 150 units / ml, 500 units / ml, 2000 units / ml, and 6000 units / ml, respectively. The solutions were left to stand at 25°C for 15 min and 1 hr. The hyaluronidase activity, ertapenem content, and appearance of the samples were measured at each sampling point. Here, both the hyaluronidase activity and ertapenem content were detected by the method described in "HPLC Measurement of Ertapenem Content and Related Substances for Injection."

[0176] Tables 28-29 show the changes in ertapenem content (labeled composition of 90-110% is acceptable) and hyaluronidase activity under different conditions for each group.

[0177] As a result, after reconstitution, all of the different formulation solutions were clear and transparent. After 15 minutes, the ertapenem content and hyaluronidase activity were stable in the different formulations. After 1 hour, the ertapenem content was within the range of 90-110% in the different formulations. Hyaluronidase activity in groups A and B decreased slightly, while hyaluronidase activity in groups C, D, E, F, and G was maintained relatively well.

[0178] [Table 28] [Table 29] Example 19. Subcutaneous polymyxin B dried powder formulation A subcutaneous polymyxin B dry powder formulation was prepared by mixing freeze-dried hyaluronidase dry powder (formulations A-G in Table 14) with polymyxin dry powder B. The mixing specifications were such that each portion contained 500,000 units of the polymyxin B active ingredient, and the hyaluronidase content was 300 units, 1,000 units, 4,000 units, and 12,000 units, respectively, depending on the combination.

[0179] After mixing, the dried powder was taken and dissolved in 2 ml of sterile water for injection, with the dried powder containing 500,000 units of polymyxin B active ingredient being dissolved. After dissolution, the hyaluronidase concentrations corresponded to 150 units / ml, 500 units / ml, 2000 units / ml, and 6000 units / ml, respectively. The solutions were left to stand at 25°C for 15 min and 1 hr. The hyaluronidase activity, polymyxin B content, and appearance of the samples were measured at each sampling point. Here, both the hyaluronidase activity and polymyxin B content were detected according to the methods of the Chinese Pharmacopoeia.

[0180] Tables 30 and 31 show the changes in polymyxin B content (labeled composition of 90-110% is eligible) and hyaluronidase activity under different conditions for each set, respectively.

[0181] As a result, after reconstitution, all of the different formulation solutions were clear and transparent. After 15 minutes, the polymyxin B content was stable in the different formulations, and the hyaluronidase activity was also stable. After 1 hour, the polymyxin B content was within the range of 90-110% in the different formulations. Hyaluronidase activity in groups A and B decreased slightly, while hyaluronidase activity in groups C, D, E, F, and G was maintained relatively well.

[0182] [Table 30] [Table 31] Example 20. Subcutaneous administration of recombinant human hyaluronidase and antibiotics. This study investigated the pharmacokinetics of subcutaneous administration of recombinant human hyaluronidase and antibiotics in mice, and how it differed from conventional intravenous administration. Ceftriaxone, ceftazidime, ertapenem, and teicoplanin were selected as model antibiotics. A total of 48 ICR male mice were selected for each antibiotic model, and divided into two groups of 24 mice each: one group administered via tail vein injection, and the other group administered subcutaneously with recombinant human hyaluronidase. A single dose of 100-200 mg / kg was administered, and the drugs were prepared and administered according to the prescription in each group. Blood samples were collected from each group at 5 min, 15 min, 30 min, 45 min, 1 h, 2 h, 4 h, and 8 h after administration. The concentration of each model antibiotic in the samples was detected using LC-MS, and pharmacokinetic parameters were calculated.

[0183] Table 32 shows the pharmacokinetic status of each model antibiotic when administered intravenously via tail vein injection into mice and subcutaneously with recombinant human hyaluronidase.

[0184] As a result, compared to intravenous injection, subcutaneous administration of recombinant human hyaluronidase yields a longer Tmax and lower Cmax, resulting in a longer duration of antibiotic action in the body, improved safety, and enhanced therapeutic efficacy. Compared to conventional subcutaneous administration, subcutaneous administration of recombinant human hyaluronidase has a higher drug absorption efficiency. This is the most common route of administration.

[0185] [Table 32] Example 21. Advantages of combined intradermal or subcutaneous administration of recombinant human hyaluronidase and antibiotics. Ceftriaxone, ceftazidime, ertapenem, and teicoplanin were selected as model antibiotics, and the effects of oral administration of these antibiotics and co-administration of recombinant human hyaluronidase subcutaneously on the intestinal microbiota of mice were investigated.

[0186] For each model antibiotic, 12 SPF-grade C57BL / 6 mice were selected and divided into two groups of 6 mice each: an oral administration group and a combined recombinant human hyaluronidase subcutaneous administration group. Additionally, 6 blank control mice were selected. The oral administration group received the model antibiotic solution orally for 5 consecutive days, while the combined recombinant human hyaluronidase subcutaneous administration group received the same amount of the model antibiotic subcutaneously for 5 consecutive days. After 5 consecutive days of administration, fecal samples were collected from each mouse group on days 1, 2, 3, 4, 5, 6, 8, 10, and 14, frozen and stored at -80°C, and prepared for detection of microbial diversity. As shown in Table 33, DNA extraction, PCR amplification, and gene sequencing were performed on the fecal samples from each group using 16S rDNA amplification and sequencing methods. The changes in the diversity of intestinal bacteria in each mouse group were then analyzed, and the relative species diversity of the experimental groups was calculated using the daily OUT values ​​of the blank control group as a baseline.

[0187] As a result, the change in gut microbiota diversity in mice administered subcutaneously with recombinant human hyaluronidase was smaller than in mice administered orally, and the gut microbiota abnormality status in mice administered subcutaneously with recombinant human hyaluronidase was better than in mice administered orally. Subcutaneous administration of recombinant human hyaluronidase in combination with antibiotics can reduce the adverse effects on the gut microbiota during antibiotic treatment and protect the diversity of gut microbiota compared to conventional oral administration of antibiotics.

[0188] [Table 33] While the applicant has illustrated the present invention by the above examples to illustrate the detailed methods of the present invention, the present invention is not limited to the above detailed methods, that is, it is not meant that the present invention must necessarily be carried out in accordance with the above detailed methods. To those skilled in the art, it will be clear that any modifications to the present invention, equivalent substitutions of the raw materials of the product of the present invention and addition of auxiliary components, and selection of specific methods are all within the scope of protection and disclosure of the present invention.

Claims

1. (a) an antibiotic selected from (i) ceftriaxone at a concentration of 250 mg / mL, (ii) ceftazidime at a concentration of 200 mg / mL, (iii) ertapenem at a concentration of 312.5 mg / mL, or (iv) teicoplanin at a concentration of 166.7 mg / mL, (b) Hyaluronidase with a concentration of 150 units / mL to 6000 units / mL, (c) A pharmaceutical composition for subcutaneous administration comprising a pharmaceutically acceptable additive containing 10 mM phosphate buffer, 10 mM methionine, 0.02% (w / v) polysorbate 20, 25 mM trehalose, and 220 mM mannitol.

2. C of the antibiotic in the composition ｍａｘ The amount of the antibiotic in the composition is increased by at least 10% compared to administration of the antibiotic alone, and / or the amount of the antibiotic in the composition is increased by at least 10%. ｍａｘ The administration time is shortened by at least 20% compared to administration of the antibiotic alone, and / or the AUC of the antibiotic in the composition. ｌａｓｔ Compared to administration of the antibiotic alone, the dose increased by at least 5%, or at least 10%. The pharmaceutical composition according to claim 1, wherein, after subcutaneous administration of the composition, the amount of reduction in the diversity of the intestinal flora compared to before administration is 90% or less of the amount of reduction after oral administration of an antibiotic compared to before administration.

3. The pharmaceutical composition according to claim 1, which is a liquid formulation.

4. A kit comprising the pharmaceutical composition described in claim 1, wherein the antibiotic and the hyaluronidase kit are mixed and packaged together.

5. A method for preparing the kit described in claim 4, (a) the step of providing an antibiotic, (b) A step of providing hyaluronidase, (c) A step of providing a pharmaceutically acceptable additive, (d) A step of mixing the additive and the hyaluronidase to prepare a freeze-dried preparation, (e) A step of preparing the antibiotic into a lyophilized formulation, A kit preparation method comprising the step of (f) mixing the lyophilized formulations from steps (d) and (e) to prepare a liquid formulation.

6. An injection system comprising a delivery device selected from the group consisting of a syringe, an infusion pump, an injection pen, a needleless device, and the injection system being filled with the pharmaceutical composition described in claim 1.

7. The pharmaceutical composition according to claim 1, used for the treatment of a disease selected from the group consisting of infections by bacteria, fungi, actinomycetes, mycoplasma, chlamydia, spirochetes, amoebas, and diseases resulting from said infection.

8. The pharmaceutical composition according to claim 7, wherein the disease is chronic osteomyelitis caused by Staphylococcus aureus, endocarditis caused by drug-resistant Staphylococcus aureus or enterococcus, typhoid fever, paratyphoid fever, bacterial food poisoning, bacterial infectious diarrhea, cholera, bacterial dysentery, brucellosis, plague, anthrax, diphtheria, pertussis, scarlet fever, epidemiological spinal corditis, tuberculosis, bacterial bloodstream infection, bacterial upper respiratory tract infection, bacterial lower respiratory tract infection, bacterial urinary tract infection, bacterial abdominal infection, dermatophytosis, fungal stomatitis, candidal vaginitis, fungal pneumonia, fungal urinary tract infection, fungal bacteremia, cryptococcosis, candidiasis, aspergillosis, or Pneumocystis.

9. The kit according to claim 4, for use in treating a disease selected from the group consisting of infections by bacteria, fungi, actinomycetes, mycoplasma, chlamydia, spirochetes, amoebas, and diseases resulting from said infection.

10. The kit according to claim 9, wherein the disease is chronic osteomyelitis caused by Staphylococcus aureus, endocarditis caused by drug-resistant Staphylococcus aureus or enterococcus, typhoid fever, paratyphoid fever, bacterial food poisoning, bacterial infectious diarrhea, cholera, bacterial dysentery, brucellosis, plague, anthrax, diphtheria, pertussis, scarlet fever, epidemic meningitis, tuberculosis, bacterial bloodborne infection, bacterial upper respiratory tract infection, bacterial lower respiratory tract infection, bacterial urinary tract infection, bacterial abdominal infection, dermatophytosis, fungal stomatitis, candidal vaginitis, fungal pneumonia, fungal urinary tract infection, fungemia, cryptococcosis, candidiasis, aspergillosis, or pneumocystis.

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