Pharmaceutical combinations for subcutaneous administration and use thereof

Abstract

The present disclosure relates to a pharmaceutical combination for subcutaneous administration comprising a hyaluronan-degrading enzyme and an antibody drug conjugate and uses thereof. The pharmaceutical combination can enable administration of larger volumes, larger doses without an increase in side effects.

Description

Drug combination for subcutaneous administration and its application

[0001] Priority Declaration

[0002] This disclosure claims priority to patent application number PCT / CN2024 / 076995, filed on February 8, 2024, entitled "A Subcutaneous Antibody Drug Conjugate," and patent application number PCT / CN2024 / 112586, filed on August 16, 2024, entitled "Drug Combination for Subcutaneous Administration and Use Thereof," both of which are incorporated herein by reference in their entirety. Technical Field

[0003] The present disclosure relates to the field of biomedicine, and in particular to a drug combination for subcutaneous administration and applications thereof. Background Art

[0004] Existing antibody-drug conjugates are all administered via intravenous injection (IV). However, IV injections face a series of challenges in clinical practice. For example, for patients with poor venous access, IV treatment may lead to loss of treatment opportunities. Injections need to be performed in outpatient clinics, increasing the risk of hospital-acquired infections. Long-term infusions and accompanying requirements may lead to productivity losses, and will result in additional medical insurance expenses such as the consumption of hospital consumables and maintenance complications, increase the complexity of the drug configuration process, and increase the difficulty of quality and safety management. At the same time, long infusion times will also result in low bed turnover rates.

[0005] Very few cytotoxic drugs are administered subcutaneously for cancer treatment because most known anticancer cytotoxic agents cause local damage in the subcutaneous or subdermal tissues after extravasation. Chinese patent CN110352201A controls local toxicity of ADC drugs by optimizing dose and time. However, subcutaneous formulations typically require higher doses than intravenous injections, which in turn requires higher injection volumes and / or drug concentrations. Although subcutaneous formulations have been developed for a variety of FDA-approved antibodies, However, subcutaneous (sc) administration is difficult for antibody-drug conjugates (ADCs) because immune cells in the skin may respond to the cytotoxic payload and mediate off-target effects, leading to toxic effects, such as local deposition of cytotoxic substances (Lucas AT, Price LSL, Schorzman AN, et al. Factors Affecting the Pharmacology of Antibody-Drug Conjugates. Antibodies (Basel, Switzerland). 2018 Feb; 7(1): E10. DOI: 10.3390 / antib7010010.).

[0006] At the same time, the conventional subcutaneous administration volume is usually less than 2 ml, so high-concentration formulations need to be developed for ADC drugs. ADC drugs are usually more hydrophobic after conjugation and loading, which also poses challenges in the development and production of ADC drug formulations, and the development of high-concentration subcutaneous formulations is even more difficult. Due to the wide variety of substances in the conjugated products, the complexity of the samples makes the preparation of non-site-specific ADCs extremely difficult. In addition, the drugs attached to the ADC and the linkers connecting the drugs also increase the complexity of ADC preparation. When ADC drugs are conjugated, the aggregation-prone areas within the antibody skeleton are exposed or antibody aggregation is caused through drug-mediated interactions, which greatly limits the ability to formulate highly stable and high-concentration ADC drugs.

[0007] Therefore, the development of ADC drugs that can be safely and efficiently administered subcutaneously is an urgent clinical need. Summary of the Invention

[0008] Some aspects of the present disclosure provide a drug combination comprising a hyaluronan degrading enzyme and an antibody drug conjugate.

[0009] In some embodiments, the pharmaceutical combination of the present disclosure is administered subcutaneously.

[0010] In some embodiments, the antibody-drug conjugates described in the pharmaceutical combinations of the present disclosure include biologically active molecular fragments.

[0011] In some embodiments, the bioactive molecules described herein are moderately toxic bioactive molecules.

[0012] In some embodiments, the biologically active molecules described herein are topoisomerase I inhibitors.

[0013] In some embodiments, the topoisomerase I inhibitor described in the present disclosure is a camptothecin-type topoisomerase I inhibitor.

[0014] In some embodiments, the topoisomerase I inhibitor described in the present disclosure is selected from camptothecin (CPT), hydroxycamptothecin (HCPT), 9-aminocamptothecin (9-AC), 7-ethyl-10-hydroxycamptothecin (SN-38), exatecan derivative (Dxd or DX-8951 derivative), irinotecan (CPT-11), topotecan, lurtotecan, belotecan or exatecan.

[0015] In some embodiments, the topoisomerase I inhibitor described herein is selected from any one of the following:

[0016] In some embodiments, the antibody drug conjugate of the present disclosure is selected from: 9MW-2921, A-315, ACR-246, ADC-2154, AZD-9592, AZD-9829, BIO-201, BLB-01D1, BSI-04702, CUSP-06, DAN-311, DB-1303, DB-1311, GPCR-targeted Project 010, HDP-201, HLX-42, HLX-43, IBI-354, IM-1021, JSKN-033, M-9140, MABS-01, MBK-101, MBK-102, MBK-105, NV-104, OBI-902, OBI-904, OBI-90 5. PRO-1102, DS-6000, IMMU-132, SMP-190, TQB-2102, DS-8201, DS-1062, DS-7300, U3-1402, XB-033, YL-201, YL-202, ZW-191, ZW-220 or ZW-251.

[0017] In some embodiments, the hyaluronidase described in the present disclosure is a mammalian hyaluronidase.

[0018] In some embodiments, the hyaluronidase described herein comprises the catalytic domain of hyaluronidase PH-20, HYAL1, HYAL2, HYAL3, HYAL4, or HYALPS1.

[0019] In some embodiments, the hyaluronidase described in the present disclosure is a hyaluronidase selected from HuPH20, HYAL1, HYAL2, HYAL3, HYAL4, or any variant thereof, or any isoform thereof.

[0020] In some embodiments, the hyaluronidase described in the present disclosure is rHuPH20, HYAL1, PH20 variant 1, or a fragment thereof.

[0021] In some embodiments, the drug combination described in the present disclosure is selected from the group consisting of the following drug combinations: rHuPH20 and DS-8201, rHuPH20 and IMMU-132, rHuPH20 and DS-1062, rHuPH20 and DS-7300, rHuPH20 and U3-1402, and PH20 variant 1 and DS-8201.

[0022] In some embodiments, the peak blood concentration (C max (ng / mL)), which is about 5% to 20% of the peak plasma concentration of the same dose of antibody drug conjugate injected alone intravenously; preferably greater than about 5%, greater than about 8%, greater than about 10%, greater than about 12%, greater than about 14%, greater than about 16%, greater than about 18% or greater than 19%.

[0023] In some embodiments, the half-life of the drug combination of the present disclosure after subcutaneous administration is about 70% to 99% of the half-life of the same dose of the antibody drug conjugate alone after intravenous injection; preferably greater than about 70%, greater than about 80%, greater than about 85%, greater than about 90% or greater than about 95%.

[0024] In some embodiments, the exposure (AUC 0-∞ (h*ng / mL)), which is about 50% to 70% of the exposure of the same dose of antibody drug conjugate injected alone intravenously; preferably greater than about 50%, greater than about 55%, greater than about 60%, greater than about 65% or greater than about 67%.

[0025] In some embodiments, in the drug combination described in the present disclosure, the antibody drug conjugate is administered at a concentration of about 2 mg / ml to 100 mg / ml; the hyaluronidase concentration is about 100 to 5000 IU / mL; alternatively, the antibody drug conjugate is administered at a concentration of about 3 mg / mL, 4 mg / mL, 5 mg / mL, 6 mg / mL, 8 mg / mL, 10 mg / mL, 12 mg / mL, 14 mg / mL, 16 mg / mL, 18 mg / mL, 20 mg / mL, 30 mg / mL, 40mg / mL, 50mg / mL, 60mg / mL, 80mg / mL or 100mg / mL; the administration concentration of the hyaluronidase is about 100IU / mL, 150IU / mL, 200IU / mL, 500IU / mL, 1000IU / mL, 2000IU / mL, 3000IU / ml or 5000IU / mL; optionally, the administration concentration of the antibody drug conjugate is about 20mg / mL, and the administration concentration of the hyaluronidase is about 500IU / mL.

[0026] In some embodiments, in the drug combination disclosed herein, the dosage ratio of the antibody drug conjugate and hyaluronidase is about 2 mg to 100 mg: 100 to 5000 U; alternatively, the dosage ratio of the antibody drug conjugate and hyaluronidase is about 3 mg to 60 mg: 100 U to 3000 U; alternatively, the dosage of the antibody drug conjugate is 3 mg / mL, 4 mg / mL, 5 mg / mL, 6 mg / ml, 10 mg / ml, 20mg / mL, 30mg / mL, 40mg / mL, 50mg / mL, 60mg / mL; optionally, the dosage of the hyaluronidase is 100IU / mL, 150IU / mL, 200IU / mL, 500IU / mL, 1000IU / mL, 2000IU / ml or 3000IU / ml; optionally, the ratio of the antibody drug conjugate to the hyaluronidase is approximately 20mg:500IU.

[0027] In some embodiments, in the drug combination described herein, the antibody-drug conjugate is DS-8201.

[0028] In some embodiments, in the drug combination described in the present disclosure, the drug combination is: rHuPH20 and DS-8201, wherein the administration concentration of DS-8201 is about 20 mg / mL, and the administration concentration of rHuPH20 is about 500 IU / mL.

[0029] In some embodiments, in the drug combination disclosed herein, the drug combination comprises rHuPH20 and DS-8201, wherein the dosage ratio of DS-8201 to rHuPH20 is approximately 20 mg:500 IU.

[0030] Some aspects of the present disclosure provide a pharmaceutical composition for subcutaneous administration, comprising any one of the aforementioned drug combinations and a pharmaceutically acceptable excipient.

[0031] In some embodiments, in the pharmaceutical composition described in the present disclosure, the pharmaceutically acceptable excipients include one or more of a lyoprotectant, a stabilizer, a solubilizer, an osmotic pressure regulator, and a buffer.

[0032] In some embodiments, in the pharmaceutical composition described in the present disclosure, the pharmaceutically acceptable excipients include one or more of sucrose, L-histidine, L-histidine hydrochloride monohydrate, polysorbate, phosphate, methionine, trehalose, mannitol and sodium chloride.

[0033] In some embodiments, in the pharmaceutical composition described in the present disclosure, the pharmaceutically acceptable excipients include one or more of sucrose, L-histidine, L-histidine hydrochloride monohydrate, and polysorbate 80.

[0034] In some embodiments, in the pharmaceutical composition of the present disclosure, the pharmaceutically acceptable excipient further comprises one or more of disodium hydrogen phosphate, methionine, trehalose, mannitol and polysorbate 20.

[0035] Some aspects of the present disclosure provide a method for treating a tumor, comprising administering any one of the aforementioned drug combinations or the aforementioned pharmaceutical composition of the present disclosure to a subject in need thereof.

[0036] In some embodiments, the tumor is lymph node metastatic cancer, mammary ductal carcinoma, breast cancer, gastric cancer, ovarian cancer, or pancreatic cancer.

[0037] Some aspects of the present disclosure provide use of any one of the aforementioned drug combinations or the aforementioned pharmaceutical composition in the preparation of a medicament for treating tumors.

[0038] Some aspects of the present disclosure provide a method for improving the bioavailability of any of the aforementioned drug conjugates, the method comprising administering any of the aforementioned hyaluronidase to a subject simultaneously with or before administering the drug conjugate.

[0039] Some aspects of the present disclosure provide a method for alleviating local irritation caused by subcutaneous administration of any of the aforementioned drug conjugates, the method comprising administering any of the aforementioned hyaluronidase to a subject simultaneously with or before administering the drug conjugate. BRIEF DESCRIPTION OF THE DRAWINGS

[0040] FIG1 is the PK experimental results of ADC+PH20 in Example 1. DETAILED DESCRIPTION

[0041] 1. Detailed Description of the Invention

[0042] Unless otherwise defined, all technical and scientific terms used in this disclosure have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. The terminology used in this detailed description is for the purpose of describing particular embodiments only and is not intended to be limiting.

[0043] All publications, patent applications, patents, GenBank or other accession numbers and other references mentioned herein are incorporated by reference in their entirety.

[0044] It is specifically contemplated that the various features described herein can be used in any combination unless the context dictates otherwise.

[0045] The following terms are used in this specification and the appended claims:

[0046] The singular forms "a," "an," and "the" are intended to include the plural forms as well, unless the context clearly indicates otherwise.

[0047] In addition, when referring to a measurable value (e.g., the amount of length of a polynucleotide or polypeptide sequence, dosage, time, temperature, etc.), the term "about" as used herein is intended to encompass variations of ±10%, ±5%, ±1%, ±0.5%, or even ±0.1% of the specified amount.

[0048] Additionally, as used herein, "and / or" refers to any one of the listed items selected in an alternative manner ("or"), and includes any and all possible combinations of two or more of the related listed items.

[0049] Unless otherwise indicated herein, numerical ranges herein are intended to describe the combination of each individual value falling within the numerical range, and each individual value should be considered to be individually recited herein. For example, if a concentration range is recited as 1% to 50%, it is intended that values ​​such as 2% to 40%, 10% to 30%, or 1% to 3% are explicitly recited in this specification. These numerical ranges or individual values ​​are only examples of what is specifically intended, and all individual values ​​between the lowest and highest values ​​recited, including the lowest and highest values, and all possible combinations of these values ​​are considered to be explicitly recited in this disclosure.

[0050] The term "immunoglobulin" refers to a class of globulins present in human and animal blood (serum), tissue fluid, and other exocrine fluids that have antibody activity and a chemical structure similar to that of antibody molecules. In some embodiments, immunoglobulins can be interpreted as antibodies, such as IgG, IgE, IgM, IgD, or IgA antibodies.

[0051] As used herein, the term "antibody" refers to a polypeptide comprising at least a light chain or heavy chain immunoglobulin variable region that specifically recognizes and binds to an antigen. The term encompasses various antibody structures, including but not limited to monoclonal antibodies, polyclonal antibodies, single-chain antibodies or multi-chain antibodies, monospecific or multispecific antibodies (e.g., bispecific antibodies, trispecific, or more multispecific), fully human antibodies, chimeric antibodies, or humanized antibodies, full-length antibodies, and antibody fragments, as long as they exhibit the desired antigen-binding activity.

[0052] The term "antigen-binding fragment" of an antibody (used interchangeably herein with "antibody fragment" and "antigen-binding portion") refers to a non-complete antibody molecule that comprises a portion of an intact antibody that is used to bind to the antigen to which the intact antibody binds. As will be appreciated by those skilled in the art, the antigen-binding portion of an antibody typically comprises amino acid residues from a "complementarity determining region" or "CDR." Antigen-binding fragments can be prepared by recombinant DNA technology, or by enzymatic or chemical cleavage of intact antibodies. Antigen-binding fragments include, but are not limited to, Fab, scFab, Fab', F(ab')2, Fab'-SH, Fv, single-chain Fv, diabody, triabody, tetrabody, minibody, and single-domain antibody VHH. For a more detailed description of antibody fragments, see: Fundamental Immunology, WE Paul, ed., Raven Press, NY (1993); Shao Rongguang et al. (eds.), Antibody Drug Research and Application, People's Medical Publishing House (2013); Hollinger et al., PNAS USA 90:6444-6448 (1993); Hudson et al., Nat. Med. 9:129-134 (2003).

[0053] In the present disclosure, antibodies include murine, chimeric, humanized or fully human antibodies prepared using techniques well known to those skilled in the art. Recombinant antibodies, such as chimeric and humanized monoclonal antibodies, including human and non-human parts, can be obtained by standard recombinant DNA techniques, and they are all useful antibodies. A chimeric antibody is a molecule in which different parts are derived from different animal species, such as a chimeric antibody having a variable region from a mouse monoclonal antibody and a constant region from a human immunoglobulin (see, for example, U.S. Patent No. 4,816,567 and U.S. Patent No. 4,816,397, which are incorporated herein by reference in their entirety). A humanized antibody refers to an antibody molecule derived from a non-human species, having one or more complementary determining regions (CDRs) derived from a non-human species and a framework region derived from a human immunoglobulin molecule (see U.S. Patent No. 5,585,089, which is incorporated herein by reference in its entirety). These chimeric and humanized monoclonal antibodies can be prepared using recombinant DNA techniques well known in the art.

[0054] Hyaluronidases (HAases) are a class of glycosidases that degrade hyaluronic acid and some glycosaminoglycans. They are found throughout the animal kingdom and include wild-type and mutant HAases, as well as their recombinant counterparts (rHAase). Hyaluronidases vary in substrate specificity and mechanism of action.

[0055] There are three major types of hyaluronidases:

[0056] 1. Mammalian hyaluronidases (EC 3.2.1.35) are endo-β-N-acetylglucosaminidase enzymes with tetrasaccharides and hexoses as the main end products. They have hydrolytic and transglycosidase activities and can degrade hyaluronic acid and chondroitin sulfate (CS), especially C4-S and C6-S.

[0057] 2. Bacterial hyaluronidases (EC 4.2.99.1) degrade hyaluronic acid and, to varying degrees, chondroitin sulfate (CS) and dermatan sulfate (DS). They are endo-β-N-acetylglucosaminidase enzymes that act via β-elimination reactions, primarily producing disaccharide end products.

[0058] 3. Hyaluronidases (EC 3.2.1.36) from leeches, other parasites, and crustaceans are endo-β-glucuronidases that hydrolyze β1-3 linkages to produce tetrasaccharide and hexose end products.

[0059] Mammalian hyaluronidases can be further divided into two groups: neutrally active and acidically active enzymes. There are six hyaluronidase-like genes (HYAL) in the human genome: HYAL1, HYAL2, HYAL3, HYAL4, HYALP1, and PH20 / SPAM1. HYALP1 is a pseudogene, and HYAL3 has not been shown to have enzymatic activity on any known substrate. HYAL4 is a chondroitinase that lacks activity on hyaluronic acid. HYAL1 is the prototype acidically active enzyme, and PH20 is the prototype neutrally active enzyme. Acidically active hyaluronidases, such as HYAL1 and HYAL2, lack catalytic activity at neutral pH. For example, HYAL1 has no catalytic activity above pH 4.5 in vitro (Frost et al., Anal Biochemistry, 1997). HYAL2 is an acidically active enzyme with very low specific activity in vitro.

[0060] Hyaluronidase-like enzymes can also be characterized as those that are locked to the plasma membrane via a glycosylphosphatidylinositol anchor. However, there are variations between species: for example, bovine PH20 is very loosely attached to the plasma membrane and is not anchored by a phospholipase-sensitive anchor (Lalancette et al., Biol Reprod. 2001 Aug; 65(2): 628-36). This unique feature of bovine hyaluronidase makes it possible to use soluble bovine testicular hyaluronidase as an extract for clinical applications. Other PH20s are lipid-anchored enzymes that are insoluble without the application of detergents or lipases. For example, human PH20 is anchored to the plasma membrane via a GPI anchor. Attempts have been made to prepare human PH20 DNA constructs that do not introduce lipid anchors, but what is obtained is an enzyme without catalytic activity, or an insoluble enzyme (Arming et al., Eur J Biochem. 1997 Aug 1; 247 (3): 810-4). It was found that naturally occurring macaque sperm hyaluronidase can be both soluble and membrane-bound. The 64kDa membrane-bound form has enzymatic activity at pH 7.0, while the 54kDa form is only active at pH 4.0 (Cherr et al., Dev Biol. 1996 Apr 10; 175 (1): 142-53). Therefore, the soluble form of PH20 lacks enzymatic activity under neutral conditions. Examples of hyaluronidases include, but are not limited to, hyaluronidases disclosed in WO2020022791A1 and WO2013102144A2.

[0061] In one embodiment, the hyaluronidase is selected from HYAL1, HYAL2, HYAL3, HYAL4, HYALP1 and PH20 / SPAM1. For example, the amino acid sequence of the optional hyaluronidase is shown in SEQ ID NO: 1-3.

[0062] Table 1 Hyaluronidase amino acid sequence

[0063] Antibody-drug conjugates (ADCs) are a class of targeted biopharmaceuticals consisting of an antibody, a linker, and a cytotoxic drug. They are designed to couple target-specific monoclonal antibodies with highly potent cytotoxic drugs via specific linkers. ADCs typically contain the following three components: an antibody with high specificity and affinity, a linker with high stability, and a highly effective small molecule cytotoxic drug. The ADC contains: (ado-trastuzumab emtansine), (famtrastuzumab deruxtecannxki), (Disitamab vedotin), (enfortumab vedotin-ejfv), (sacituzumab govitecan hziy), (cetuximab saratolacan sodium), (tisotumab vedotin-tftv), (mirvetuximab soravtansine-gynx), Mylotarg, Adcetris, Besponsa, Lumoxiti, Polivy, Blenrep, Zynlonta, Vidicituximab; 9MW-2921 (Mabwell Shanghai Bioscience Co., Ltd), A-315 (KLUS Pharma Inc), ACR-246 (Hangzhou Adcoris Biopharmaceutical Co., Ltd), ADC-2154 (Hangzhou Adcoris Biopharmaceutical Co., Ltd), AZD-9592 (AstraZeneca Plc), AZD-9829 (AstraZeneca Plc), BIO-201 (BiOneCure Therapeutics Inc), BLB-01D1 (Bristol-Myers Squibb Co., Systimmune Inc), BSI-04702 (Biosion Inc, OBI Pharma Inc), CUSP-06 (Multitude therapeutics Inc, OnCusp Therapeutics Inc), DAN-311 (Dantari Inc), DB-1303 (BioNTech SE, Duality Biologics(Shanghai)Co., Ltd), DB-1311 (BioNTech SE, Duality Biologics(Shanghai)Co.Ltd),GPCR-targeted Project 010(Protheragen Inc),HDP-201(Heidelberg PharmaAG),HLX-42(Shanghai Henlius,Biotech Inc),HLX-43(Shanghai Henlius Biotech Inc),IBI-354(InnoventBiologics Inc),IM-1021(Immunome Inc),JSKN-033(Alphamab Oncology),M-9140(Merck KGaA),MABS-01(Manhattan Biosolutions Inc),MBK-101(Mablink Bioscience SAS),MBK-102(Mablink Bioscience SAS),MBK-105(Mablink Bioscience SAS),Monoclonal Antibody Conjugate to Inhibit TOP1 and CD155 for Unspecified Cancer(Tasrif Pharmaceutical Inc),Next-Generation ADCs(Inxmed(Nanjing)Co.Ltd),NV-104(Nano Valent Pharmaceuticals Inc(Inactive)),OBI-902(OBI Pharma Inc),OBI-904(OBI Pharma Inc),OBI-905(OBI Pharma Inc),PRO-1102(ProfoundBio Suzhou Co.Ltd),raludotatug deruxtecan(Daiichi Sankyo Co.Ltd,DS-6000),sacituzumab govitecan(Immunomedics,IMMU-132),SMP-190(Xiling Lab Co.Ltd),TQB-2102(Chia Tai Tianqing Pharmaceutical Group Co.Ltd),trastuzumab deruxtecan(Daiichi Sankyo Co.Ltd,AstraZeneca,DS-8201),datopotamab deruxtecan(Daiichi Sankyo Co.Ltd, AstraZeneca, DS-1062), Ifinatamab deruxtecan (Daiichi Sankyo Co. Ltd, DS-7300), Patritumab deruxtecan (Daiichi Sankyo Co. Ltd, Merck & Co., Inc., U3-1402), XB-033 (Exelixis Inc), YL-201 (Suzhou Medilink Therapeutics Ltd), YL-202 (Suzhou Medilink Therapeutics Ltd), ZW-191 (Zymeworks Inc), ZW-220 (Zymeworks Inc), ZW-251 (Zymeworks Inc). .

[0064] drug

[0065] As used herein, "drug" refers to any compound having a desired biological activity and a reactive functional group for preparing the conjugates described in the present disclosure. The desired biological activity includes diagnosing, curing, alleviating, treating, and preventing diseases in humans or other animals. Therefore, it is necessary to have the necessary reactive functional groups. The compounds referred to by the term "drug" include drugs confirmed in official national pharmacopeias, as well as drugs such as the official U.S. Homeopathic Pharmacopoeia, the official National Formulary, or any supplements thereto. Typical drugs are listed in the Physician's Desk Reference (PDR) and the Orange Book of the U.S. Food and Drug Administration (FDA). It should be understood that as new drugs are constantly discovered and developed, these drugs should also be included in the "drug" of the conjugate drugs described in the present disclosure.

[0066] Drugs that can be used to construct the ADC of the present disclosure include, but are not limited to, cytotoxic drugs and non-cytotoxic drugs.

[0067] The term "cytotoxic drug" is equivalent to "cytotoxic agent". "Effective load" refers to a substance that inhibits cell expression activity, disrupts cell function and / or causes cell damage. The term includes radioactive isotopes, chemotherapeutic agents, and toxins, such as small molecule toxins or enzymatically active toxins of bacterial, fungal, plant or animal origin, including fragments and / or variants thereof. Examples of cytotoxic drugs include, but are not limited to: dolastatin (such as auristatin), camptothecin and its derivatives, calicheamicin, doxorubicin, duocarmycin, hemipterafil, IGN, PBD, amanitine, Clezutoclax, diphtheria toxin, eribulin, irinotecan, ricin, saporin, Azonafide, Amberstatin269, dmDNA31, Clezutoclax, belotecan, vinblastine, DM21C, Nigrin, glycosides (such as cardiac glycosides), etc.

[0068] The term "moderate toxicity" usually refers to IC 50 The toxicity value is at the nanomolar (0.1-10 nM) level. Common reagents with moderate toxicity include auristatin derivatives such as MMAE / MMAF, camptothecin derivatives such as DXd and SN-38, etc.

[0069] Non-cytotoxic drugs include: glucocorticoids, photosensitizers (such as 700DX), Bcl-xL inhibitors, NAMPT inhibitors, TLR7 / 8 agonists, STING agonists, radiolabeled elements (such as iodine, lutetium, yttrium, indium), etc.

[0070] In certain aspects, cytotoxic agents are further classified as: microtubule inhibitors, DNA damaging agents, topoisomerase 1 inhibitors, etc. Further, microtubule inhibitors include dolastatins, such as MMAE and MMAF; maytansines, such as maytansine, DM1, and DM4; and halichondrins, such as halichondrin B, eribulin, and eribulin mesylate. DNA damaging agents include calicheamicin and pyrrolobenzodiazepine (PBD). Topoisomerase 1 inhibitors include camptothecins.

[0071] Table 2 Commonly used cytotoxic drugs

[0072] Camptothecins are topoisomerase I inhibitors. Camptothecins are currently the most effective and promising class of alkaloid anti-tumor drugs. Camptothecin molecules typically contain five rings: A, B, C, D, and E. Rings A and B are quinoline rings, ring C is a pyrrole ring, ring D is a pyridone, and ring E is an α-hydroxy lactone with an S-type chiral carbon.

[0073] Common camptothecin chemicals include camptothecin (CPT), hydroxycamptothecin (HCPT), 9-aminocamptothecin (9-AC), 7-ethyl-10-hydroxycamptothecin (SN-38), exatecan derivative (Dxd or DX-8951 derivative), irinotecan (CPT-11), topotecan, lurtotecan, belotecan, and exatecan. Exemplary camptothecin-type pentazocane isomerase I inhibitors also include camptothecin and its derivatives disclosed in CN117398474 A, for example:

[0074] The "linkers" described in the present disclosure can be divided into two categories according to the mechanism of drug release in cells: non-cleavable linkers and cleavable linkers.

[0075] For antibody-drug conjugates containing non-cleavable linkers, the drug release mechanism is as follows: after the conjugate binds to the antigen and is internalized by the cell, the antibody is enzymatically hydrolyzed in the lysosome, releasing the active molecule composed of the small molecule drug, the linker, and the antibody amino acid residues. The resulting structural change in the drug molecule does not reduce its cytotoxicity, but because the active molecule is charged (amino acid residues), it cannot penetrate neighboring cells. Therefore, such active drugs cannot kill neighboring tumor cells that do not express the target antigen (antigen-negative cells) (bystander effect).

[0076] Cleavable linkers, as the name suggests, can cleave within target cells and release the active drug (the small molecule drug itself). Cleavable linkers can be divided into two main categories: chemically labile linkers and enzyme-labile linkers. Chemically labile linkers can selectively cleave due to differences in plasma and cytoplasmic properties, such as pH and glutathione concentration. pH-sensitive linkers are often referred to as acid-cleavable linkers. These linkers are relatively stable in the neutral environment of blood (pH 7.3–7.5), but are hydrolyzed within the weakly acidic endosomes (pH 5.0–6.5) and lysosomes (pH 4.5–5.0). First-generation antibody-drug conjugates (ADCs) primarily utilize this type of linker, such as hydrazones, carbonates, acetals, and ketals. Due to the limited plasma stability of acid-cleavable linkers, ADCs based on these linkers typically have a short half-life (2–3 days). This short half-life has, to some extent, limited the application of pH-sensitive linkers in newer ADCs.

[0077] Glutathione-sensitive linkers, also known as disulfide linkers, trigger drug release based on the difference between the high intracellular glutathione concentration (millimolar range) and the relatively low glutathione concentration (micromolar range) in blood. This is particularly true for tumor cells, where low oxygen levels lead to increased reductase activity, resulting in higher glutathione concentrations. Disulfide bonds are thermodynamically stable, resulting in good stability in plasma.

[0078] Enzyme-labile linkers, such as peptide linkers, enable better control of drug release. Peptide linkers can be effectively cleaved by lysosomal proteases, such as cathepsin B or plasmin (the levels of such enzymes are increased in some tumor tissues). This peptide linkage is considered very stable in the plasma circulation because the unfavorable extracellular pH and serum protease inhibitors generally render the proteases inactive. Due to their high plasma stability and good intracellular cleavage selectivity and efficiency, enzyme-labile linkers are widely used as cleavable linkers in antibody-drug conjugates. Typical enzyme-labile linkers include Val-Cit (VC) and Phe-Lys.

[0079] Suicide linkers are typically embedded between a cleavable linker and an active drug, or are themselves part of the cleavable linker. The mechanism of action of a self-releasing linker is that, upon cleavage of the cleavable linker under appropriate conditions, the self-releasing linker spontaneously rearranges its structure, releasing the attached active drug. Common suicide linkers include p-aminobenzyl alcohol (PAB) and β-glucuronide.

[0080] Pharmaceutical compositions and methods of administration

[0081] As used herein, a "pharmaceutical composition" refers to a formulation or combination of formulations containing one, two, or more active ingredients, which allows the active ingredients contained therein to exist in a biologically effective form and does not contain additional ingredients that are unacceptably toxic to the subject to which the formulation is administered. When a "pharmaceutical composition" is present as a combination of separate formulations containing two or more different active ingredients, the formulations may be administered simultaneously, sequentially, separately, or at intervals, with the goal of leveraging the biological activities of the multiple active ingredients for combined treatment of a disease.

[0082] The pharmaceutical composition disclosed herein comprises at least the following (1) to (3) components in combination:

[0083] (1) Hyaluronidase;

[0084] (2) ADC drugs;

[0085] (3) Preparations containing one, two or more active ingredients.

[0086] Another aspect of the present disclosure relates to a method of treating cancer in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a composition of the present disclosure, thereby treating the cancer.

[0087] Conventional methods, known to those of ordinary skill in the medical field, can be used to administer the pharmaceutical composition to a subject, depending on the type of disease to be treated or the location of the disease. This composition can also be administered by other conventional routes, for example, orally, parenterally, by inhalation spray, topical administration, rectal administration, nasal, oral, vaginal or by implantation. The term "parenteral" as used herein includes injection or infusion techniques at sites such as subcutaneous, intracutaneous, intravenous, intramuscular, intraarticular, intraarterial, intrasynovial, intrasternal, intrathecal, intralesional and intracranial. In some cases, subcutaneous administration can be selected as the mode of administration. The subcutaneous administration may or may not include the administration of hyaluronidase. Wherein hyaluronidase can be administered before, after or simultaneously with the ADC drug administration. As used herein, the term "simultaneously" refers to being close enough in time to produce a combined effect (i.e., simultaneously, it can be simultaneous, or it can be two or more events that occur within a short period of time before or after each other).

[0088] In some embodiments, it may be necessary to administer the compositions of the present disclosure to a subject more than once to provide a therapeutic or other beneficial effect. The compositions of the present disclosure may be administered, for example, 1, 2, 3, 4, or more times.

[0089] In some embodiments, compositions disclosed herein also include at least one pharmaceutically acceptable carrier, excipient and / or medium, for example, solvent, buffer, solution, dispersion medium, coating, antibacterial and antifungal agent, isotonic agent and absorption delay agent. In some embodiments, pharmaceutically acceptable carrier, excipient and / or medium may include saline, buffered saline, glucose, water, glycerol, sterile isotonic aqueous buffer, phosphate buffer solution, amino acid buffer, bicarbonate buffer solution and combination thereof. In some embodiments, pharmaceutically acceptable carrier, excipient and / or medium include phosphate buffered saline, sterile saline, lactose, sucrose, calcium phosphate, dextran, agar, pectin, peanut oil, sesame oil, medical grade mannitol, lactose, starch, magnesium stearate, saccharin sodium, cellulose, magnesium carbonate, polyol (for example, glycerol, propylene glycol and liquid polyethylene glycol, etc.) or its suitable mixture. In some embodiments, compositions disclosed herein also include a small amount of emulsifying or wetting agent or pH buffer. Formulations of the compositions disclosed herein can be prepared for storage by mixing with physiologically acceptable carriers, excipients, or stabilizers in the form of, for example, lyophilized powders, slurries, aqueous solutions, or suspensions.

[0090] In some embodiments, the composition is in solid form such as a lyophilized powder, liquid solution, suspension, emulsion, tablet, pill, capsule, sustained release formulation or powder suitable for reconstitution. In some embodiments, the composition can be formulated for delivery using liposomes, nanocapsules, microparticles, microspheres, lipid particles, vesicles, nanospheres, nanoparticles, etc.

[0091] When using the antibody-drug conjugates of the present disclosure for treatment, delivery can be performed by conventional methods in the art. For example, it can be introduced into cells using liposomes, hydrogels, cyclodextrins, biodegradable nanocapsules, or bioadhesive microspheres. Alternatively, the nucleic acid or vector can be delivered locally by direct injection or by using an infusion pump. Other methods include various transport and carrier systems using conjugates and biodegradable polymers.

[0092] Unless otherwise indicated, the term "polypeptide" as used herein includes peptides and proteins.

[0093] The term "nucleotide" or "polynucleotide" means deoxyribonucleotides, deoxyribonucleosides, ribonucleosides or ribonucleotides and polymers thereof in single or double stranded form. Unless specifically limited, the term encompasses nucleic acids containing known analogs of natural nucleotides, which have binding properties similar to reference nucleic acids and are metabolized in a manner similar to naturally occurring nucleotides. Unless specifically limited otherwise, the term also means oligonucleotide analogs, which include PNA (peptide nucleic acid), DNA analogs used in antisense technology (phosphorothioate, phosphoramidate, etc.). Unless otherwise specified, a specific nucleic acid sequence also implicitly encompasses conservatively modified variants thereof (including but not limited to degenerate codon substitutions) and complementary sequences as well as explicitly specified sequences. In particular, degenerate codon substitutions can be achieved by generating sequences in which position 3 of one or more selected (or all) codons is substituted with mixed-base and / or deoxyinosine residues (Batzer et al., Nucleic Acid Res. 19:5081 (1991); Ohtsuka et al., J. Biol. Chem. 260:2605-2608 (1985); and Cassol et al., (1992); Rossolini et al., Mol Cell. Probes 8:91-98 (1994)).

[0094] The terms "polypeptide" and "protein" are used interchangeably herein to refer to a polymer of amino acid residues. That is, a description directed to a polypeptide applies equally to describing a peptide and describing a protein, and vice versa. The terms apply to naturally occurring amino acid polymers as well as amino acid polymers in which one or more amino acid residues is a non-naturally encoded amino acid. As used herein, the terms encompass amino acid chains of any length, including full-length proteins (i.e., antigens), in which the amino acid residues are linked via covalent peptide bonds.

[0095] The term "host cell" means a cell comprising a nucleic acid of the present disclosure, regardless of the method used for insertion to produce a recombinant host cell, such as direct uptake, transduction, mating, or other methods known in the art. The exogenous polynucleotide may be maintained as a non-integrating vector, such as a plasmid, or may be integrated into the host genome. The host cell may be a prokaryotic cell or a eukaryotic cell.

[0096] The term "transformation" refers to a process by which a heterologous DNA sequence is introduced into a host cell or organism.

[0097] The term "expression" means the transcription and / or translation of an endogenous gene or a transgene in a cell.

[0098] The beneficial effects of the present disclosure include at least: the subcutaneous ADC composition of the present disclosure enables administration in larger volumes and larger doses. Furthermore, the combination of subcutaneous ADC and hyaluronidase results in higher peak plasma concentrations, a shorter time to peak concentration, a longer half-life, and higher exposure compared to administration of ADC alone. Furthermore, the subcutaneous ADC composition of the present disclosure exhibits fewer side effects (reduced number of subjects experiencing erythema and skin edema), achieving a superior therapeutic effect.

[0099] II. Embodiment

[0100] The technical solutions of the present disclosure are further illustrated below by specific embodiments. Those skilled in the art should understand that the embodiments described herein are merely intended to aid understanding of the present disclosure and should not be construed as limiting the present disclosure. Experimental methods in the following examples, where specific conditions are not specified, were performed according to conventional methods and conditions, or as selected from commercial product specifications.

[0101] Example 1 Preparation of recombinant human hyaluronidase (rHuPH20) protein

[0102] CHO cells were cultured in suspension in serum-free medium and fed-batch controlled culture was performed using serum-free feed medium. The culture was gradually expanded to a 30 L reactor scale through shake flask culture.

[0103] On day 3-4 of culture, feed medium is added daily to the bioreactor at a rate of 2%-5% of the actual culture volume. The culture temperature is maintained at 35-37°C, and the pH is maintained at 7.0 by adding 10% (w / v) Na2CO3 and CO2. The reactor aeration rate is controlled at 0.015-0.15 vvm, the rotational speed is controlled at 80-150 rpm, and the dissolved oxygen level is controlled at 20-40%. During the cell culture process, samples are taken daily to monitor temperature, pH, glucose concentration, lactate concentration, osmotic pressure, and protein expression. When the CHO cell viability falls below 80% or the culture period reaches 14-20 days, the culture is terminated to obtain recombinant human hyaluronidase (rHuPH20).

[0104] The obtained recombinant human hyaluronidase supernatant was subjected to chromatography and filtration to purify the protein, obtaining a recombinant human hyaluronidase stock solution with a SEC purity greater than 95% and a hyaluronidase with a RP purity greater than 85%. The amino acid sequence of recombinant human hyaluronidase (rHuPH20) is shown in SEQ ID NO: 1.

[0105] Example 2 Subcutaneous injection of recombinant human hyaluronidase (rHuPH20) and DS-8201

[0106] This experiment used the recombinant human hyaluronidase (rHuPH20) prepared in Example 1 to observe the effect of subcutaneous injection of a hyaluronidase liquid preparation on subcutaneous pressure infusion of a DS8201 solution into the back of nude mice, record the infusion time, and calculate the infusion rate.

[0107] Thirty-six female BALB / c nude mice were randomly divided into three groups based on body weight: 1) negative control (normal saline + DS-8201), 2) recombinant human hyaluronidase liquid preparation group (50 units / site administered first, followed by DS8201 injection), and 3) recombinant human hyaluronidase liquid preparation + DS-8201 mixed group, with 12 animals in each group. In group 3, prior to administration, 300,000 units / ml recombinant human hyaluronic acid solution was added to DS-8201 solution at a ratio of 1:60 and mixed thoroughly before administration. After anesthesia, two symmetrical injection sites were marked on the back with a marker pen. The recombinant human hyaluronidase liquid preparation or positive control solution was administered subcutaneously on the right side of the back. A corresponding volume of normal saline was administered subcutaneously on the left side of the back as an autologous negative control. A 1 mL syringe containing a certain concentration of DS-8201 solution was connected to one end of a 0.76 mm inner diameter PE tube and a 0.55 mm needle was connected to the other end. A certain volume of DS8201 solution was subcutaneously infused at the administration site under water pressures of 20 cm, 30 cm, and 40 cm (6 animals per group at each water pressure). The infusion time t (min) was recorded and the infusion rate v (μL / min) was calculated.

[0108] Under the experimental conditions, the recombinant human hyaluronidase liquid preparation can significantly increase the rate of subcutaneous infusion of high-concentration DS-8201 solution in nude mice.

[0109] Example 3 PK study of ADC+rHuPH20

[0110] Twenty-four Sprague-Dawley rats weighing approximately 250 g were randomly divided into four groups, each consisting of six rats, half male and half female. Rats in the corresponding groups received a single tail vein injection of DS-8201 or a single subcutaneous injection of DS-8201 or rHuPH20 + DS-8201. Serum was collected by centrifugation 5 minutes, 6 hours, 12 hours, 24 hours, 2 days, 5 days, 9 days, 14 days, 21 days, and 28 days after administration, after which whole blood samples were allowed to rest for 2 hours. Serum samples were then collected and tested. Detailed grouping and design are shown in Table 3 below.

[0111] The test found that the exposure of the subcutaneous administration group (C max , AUC) were lower than those in the group without rHuPH20 addition, as shown in Table 4 and Figure 1.

[0112] Table 3 PK experimental group design results of ADC+rHuPH20

[0113] Table 4 PK experimental results of ADC+rHuPH20

[0114] As shown in Table 4, subcutaneous administration of the ADC combined with hyaluronidase increased peak plasma concentration by 46.21%, shortened time to peak concentration by 50% (24 hours), prolonged half-life by 9.7% (19.2 hours), and increased exposure by 26.4% compared to subcutaneous administration of the ADC alone. Compared to intravenous administration of the ADC alone, peak plasma concentration was approximately 18.5%, half-life was approximately 91.4%, and exposure was approximately 67.8%.

[0115] Example 4 Toxicity Experiment

[0116] Twenty-four Sprague-Dawley (SD) rats weighing approximately 250 g were randomly divided into four groups, each consisting of six rats, half male and half female. Rats in the corresponding groups received either a tail vein injection of DS-8201 or a subcutaneous injection of rHuPH20 + DS-8201 diluted solution, DS-8201, or rHuPH20 + DS-8201 via the back. Dosing was performed twice every two weeks (Day 1 and Day 15). The rats' condition and the site of administration were observed throughout the experiment. The specific grouping and design are shown in Table 5.

[0117] On D18 (72 hours after the last dose), samples from the dosing sites (skin, subcutaneous tissue, and muscle tissue) of Groups 3 and 4 animals and the blank control area (skin, subcutaneous tissue, and muscle tissue) of Group 3 animals were obtained, fixed, paraffin-embedded, sectioned, and stained with HE. Histopathological examinations were performed to assess local skin irritation in the rats. Samples were obtained bilaterally in both groups, with left and right sites distinguished.

[0118] Table 5 Toxicity experiment group design

[0119] The test results are shown in Table 6. The local symptoms after administration have different manifestations:

[0120] Table 6 Toxicity test results

[0121] Post-administration observations revealed that the animal experimental group receiving subcutaneous administration of DS-8201 with rHuPH20 had a lower incidence of erythema (reduced from 100% to 50%) compared to the group receiving direct subcutaneous administration of DS-8201, demonstrating that rHuPH20 administration reduced toxicity and side effects. Therefore, it is believed that rHuPH20 has a significant advantage in reducing local irritation.

[0122] Example 5 In vivo experiment

[0123] 5.1 In this example, MDA-MB-175 cells were used to construct an animal model for in vivo experiments on ADCs.

[0124] MDA-MB-175 cell line was cultured in culture medium, and cells in the exponential growth phase were collected. On the day of inoculation, 2×10 7 Each mouse was inoculated with 0.2 ml of PBS with Matrigel (1:1) at the third nipple pad on the right side. A 0.36 mg estrogen tablet was implanted one day before inoculation. For each model, when the average tumor volume of the tumor-bearing mice reached 250 mm 3 Start medication.

[0125] Table 7 Experimental group design

[0126] 5.2 This example uses NCI-N87 cells to construct an animal model for in vivo experiments on ADCs

[0127] NCI-N87 cell line was cultured in culture medium, and cells in the exponential growth phase were collected. On the day of inoculation, the cells were inoculated with 1×10 7 0.1 ml PBS with Matrigel (1:1) was inoculated into the subcutaneous area of ​​the right anterior back of the mouse. In each model, when the average tumor volume of the tumor-bearing mice reached about 200 mm 3 Start medication.

[0128] Table 8 Experimental group design

[0129] 5.3 This example uses an animal model constructed with SK-OV-3 cells to conduct in vivo experiments on ADC

[0130] SK-OV-3 cells were cultured in culture medium, and cells growing in the exponential phase were collected. On the day of inoculation, 1×107 0.1 ml PBS with Matrigel (1:1) was inoculated into the subcutaneous area of ​​the right front back of the mouse. In each model, when the average tumor volume of the tumor-bearing mice reached about 200-250 mm 3 Start medication.

[0131] Table 9 Experimental group design

[0132] 5.4 This example uses an animal model constructed with JIMT-1 cells to conduct in vivo experiments on ADCs

[0133] JIMT-1 cells were cultured in culture medium, and cells growing in the exponential phase were collected. On the day of inoculation, 5×10 6 0.1 ml PBS was inoculated into the subcutaneous area of ​​the right front back of the mouse. In each model, when the average tumor volume of the tumor-bearing mice reached about 200-250 mm 3 .

[0134] Table 10 Experimental group design

[0135] 5.5 This example uses an animal model constructed with Capan-1 cells to conduct in vivo experiments on ADCs

[0136] Capan-1 cells were cultured in culture medium, and cells growing in the exponential phase were collected. On the day of inoculation, 5×10 6 0.1 ml PBS with Matrigel (1:1) was inoculated into the subcutaneous area of ​​the right anterior back of each mouse. When the average tumor volume of the tumor-bearing mice reached about 170 mm 3 Start medication.

[0137] Table 11 Experimental group design

[0138] Example 6. In vivo drug efficacy experiment on lymph node metastasis

[0139] 6.1 In this example, a breast cancer lymph node metastasis model was established in NOD / SCID mice using MDA-MB-231-Luc human breast cancer cell suspension.

[0140] 8-week-old female NOD / SCID mice were treated with 3×10 6 MDA-MB-231-Luc cells were subcutaneously inoculated into the left second mammary fat pad of NOD / SCID female mice, and MDA-MB-231 orthotopic tumors were induced under inhalation anesthesia (3% sevoflurane, 100% oxygen). The technology tracks tumor growth and ALN (axillary lymph node) metastasis by bioluminescent imaging of MDA-MB-231-Luc cells. Dosing begins once lymph node metastasis is detected. Mice are administered varying doses of PBS, DS8201, T-DM1, DS8201 + rHuPH20, and T-DM1 + ​​rHuPH20 via intravenous or subcutaneous injection into the forefoot pad. Mice are then euthanized, and ALNs are harvested for FACS or H&E staining.

[0141] Table 12 Experimental group design

[0142] 6.2 In this example, a breast cancer lymph node metastasis model was established in NOD / SCID mice using MCF-7-Luc human breast cancer cell suspension.

[0143] 8-week-old female NOD / SCID mice were treated with 3×10 6 MCF-7-Luc cells were subcutaneously inoculated into the left second mammary fat pad of NOD / SCID female mice, and MCF-7 orthotopic tumors were induced under inhalation anesthesia (3% sevoflurane, 100% oxygen). The technology tracks tumor growth and ALN (axillary lymph node) metastasis by bioluminescent imaging of MCF-7-Luc cells. Dosing begins when lymph node metastasis is detected, with mice receiving varying doses of PBS, DS8201, T-DM1, DS8201 + rHuPH20, and T-DM1 + ​​rHuPH20 via intravenous or subcutaneous injection into the forefoot pad. Mice are then euthanized, and ALNs are harvested for FACS or H&E staining.

[0144] Table 13 Experimental group design

[0145] In summary, the examples disclosed herein demonstrate that the subcutaneous ADC compositions disclosed herein enable administration in larger volumes and larger doses. Furthermore, combined administration of subcutaneous ADC and hyaluronidase results in higher peak plasma concentrations, a shorter time to peak concentration, a longer half-life, and higher exposure compared to administration of ADC alone. Furthermore, administration of the subcutaneous ADC compositions disclosed herein results in fewer side effects (reduced number of subjects experiencing erythema and skin edema), achieving superior therapeutic efficacy.

Claims

1. A drug combination, wherein: The composition includes a hyaluronan degrading enzyme and an antibody drug conjugate.

2. The pharmaceutical combination according to claim 1, wherein The drug combination is administered subcutaneously.

3. The pharmaceutical combination according to claim 1 or 2, wherein The antibody-drug conjugate includes a biologically active molecule fragment.

4. The pharmaceutical combination according to claim 3, wherein The bioactive molecule is a bioactive molecule with moderate toxicity.

5. The pharmaceutical combination according to claim 3 or 4, wherein The bioactive molecule is a topoisomerase I inhibitor.

6. The pharmaceutical combination according to claim 5, wherein The topoisomerase I inhibitor is a camptothecin topoisomerase I inhibitor.

7. The pharmaceutical combination according to claim 6, wherein The topoisomerase I inhibitor is selected from camptothecin (CPT), hydroxycamptothecin (HCPT), 9-aminocamptothecin (9-AC), 7-ethyl-10-hydroxycamptothecin (SN-38), exatecan derivative (Dxd or DX-8951 derivative), irinotecan (CPT-11), topotecan, lurtotecan, belotecan or exatecan.

8. The pharmaceutical combination according to claim 6 or 7, wherein The topoisomerase I inhibitor is selected from any one of the following:

9. The pharmaceutical combination according to any one of claims 1 to 8, wherein The antibody drug conjugate is selected from: 9MW-2921, A-315, ACR-246, ADC-2154, AZD-9592, AZD-9829, BIO-201, BLB-01D1, BSI-04702, CUSP-06, DAN-311, DB-1303, DB-1311, GPCR-targeted Project 010, HDP-201, HLX-42, HLX-43, IBI-354, IM-1021, JSKN-033, M-9140, MABS-01, MBK-101, MBK-102, MBK-105, NV-104, OBI-902, OBI-904, OBI-90 5. PRO-1102, DS-6000, IMMU-132, SMP-190, TQB-2102, DS-8201, DS-1062, DS-7300, U3-1402, XB-033, YL-201, YL-202, ZW-191, ZW-220 or ZW-251.

10. The pharmaceutical combination according to any one of claims 1 to 9, wherein The hyaluronidase is a mammalian hyaluronidase.

11. The pharmaceutical combination according to any one of claims 1 to 10, wherein The hyaluronidase comprises the catalytic domain of hyaluronidase PH-20, HYAL1, HYAL2, HYAL3, HYAL4 or HYALPS1.

12. The pharmaceutical combination according to any one of claims 1 to 11, wherein The hyaluronidase is selected from HuPH20, HYAL1, HYAL2, HYAL3, HYAL4, or any variants thereof, or any isoforms thereof.

13. The pharmaceutical combination according to any one of claims 1 to 12, wherein: The hyaluronidase is any one of SEQ ID NOs: 1 to 3 or a fragment thereof.

14. The pharmaceutical combination according to any one of claims 1 to 13, wherein The drug combination is selected from the group consisting of rHuPH20 and DS-8201, rHuPH20 and IMMU-132, rHuPH20 and DS-1062, rHuPH20 and DS-7300, rHuPH20 and U3-1402, and PH20 variant 1 and DS-8201.

15. The pharmaceutical combination according to any one of claims 1 to 14, wherein The antibody drug conjugate is administered at a concentration of about 2 mg / ml to 100 mg / ml; the hyaluronidase is administered at a concentration of about 100 to 5000 IU / mL; Optionally, the antibody drug conjugate is administered at a concentration of about 2 mg / mL, 6 mg / mL, 8 mg / mL, 10 mg / mL, 20 mg / mL, 30 mg / mL, 40 mg / mL, 50 mg / mL, 60 mg / mL, 80 mg / mL or 100 mg / mL; the hyaluronidase is administered at a concentration of about 100 IU / mL, 150 IU / mL, 200 IU / mL, 500 IU / mL, 1000 IU / mL, 2000 IU / mL, 3000 IU / mL or 4000 IU / mL; Optionally, the antibody drug conjugate is administered at a concentration of about 20 mg / mL, and the hyaluronidase is administered at a concentration of about 500 IU / mL.

16. The pharmaceutical combination according to any one of claims 1 to 14, wherein The dosage ratio of the antibody drug conjugate and hyaluronidase is about 2 mg to 100 mg: 100 to 5000 IU; Optionally, the dosage ratio of the antibody drug conjugate and hyaluronidase is about 3 mg to 60 mg: 100 IU to 3000 IU; Optionally, the dosage of the antibody drug conjugate is 3 mg / mL, 4 mg / mL, 5 mg / mL, 6 mg / ml, 10 mg / ml, 20 mg / mL, 30 mg / mL, 40 mg / mL, 50 mg / mL, 60 mg / mL; Optionally, the hyaluronidase is administered in an amount of 100 IU / mL, 150 IU / mL, 200 IU / mL, 500 IU / mL, 1000 IU / mL, 2000 IU / ml or 3000 IU / ml; Optionally, the ratio of the antibody drug conjugate to hyaluronidase is about 20 mg:500 IU.

17. The pharmaceutical combination according to any one of claims 1 to 16, wherein The antibody-drug conjugate is DS-8201.

18. The pharmaceutical combination according to any one of claims 1 to 17, wherein The drug combination is: rHuPH20 and DS-8201, wherein the administration concentration of the DS-8201 is about 20 mg / mL, and the administration concentration of the rHuPH20 is about 500 IU / mL.

19. The pharmaceutical combination according to any one of claims 1 to 18, wherein The drug combination is: rHuPH20 and DS-8201, wherein the dosage ratio of DS-8201 to rHuPH20 is approximately 20 mg:500 IU.

20. A pharmaceutical composition for subcutaneous administration, wherein: The pharmaceutical composition comprises the drug combination according to any one of claims 1 to 19 and pharmaceutically acceptable excipients.

21. A method for treating tumors, wherein: The method comprises administering the pharmaceutical combination according to any one of claims 1 to 19 or the pharmaceutical composition according to claim 20 to a subject in need thereof.

22. The method of claim 21, wherein: The tumor is lymph node metastasis cancer, mammary ductal carcinoma, breast cancer, gastric cancer, ovarian cancer or pancreatic cancer.

23. Use of the pharmaceutical combination according to any one of claims 1 to 19 or the pharmaceutical composition according to claim 20 in the preparation of a medicament for treating tumors.

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