Applications of mitoxantrone hydrochloride liposomes

Abstract

To provide use of a Mitoxantrone Hydrochloride liposome in the manufacture of a medicament for treating bone and soft tissue sarcomas.SOLUTION: Use of a Mitoxantrone Hydrochloride liposome in the manufacture of a medicament for the treatment of metastatic or locally advanced bone and soft tissue sarcomas that have failed at least first line therapy is provided.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention belongs to the field of anti-tumor, and in particular relates to the use of mitoxantrone hydrochloride liposomes in the manufacture of medicaments for treating, for example, urothelial carcinoma, breast cancer, and bone and soft tissue sarcoma. [Background technology]

[0002] Urothelial carcinoma (URC) is a multifocal malignant tumor derived from the urothelium. It includes renal pelvic cancer, ureteral cancer, bladder cancer, and urethral cancer, and is the most common urological tumor. Globally, bladder-derived tumors ranked 12th in incidence among all solid tumors in 2018, accounting for 2.1% of all tumor deaths (Non-Patent Document 1). In China, bladder-derived tumors were not in the top 10 in incidence among all tumors in 2015, ranking 7th in men with a rate of 6.2 per 100,000 people, and not in the top 10 in women, ranking 14th in standardized mortality rate among all tumors with a rate of 1.96 per 100,000 people (Non-Patent Document 2). The male-to-female ratio in bladder cancer is approximately 3:1 (Non-Patent Document 1).

[0003] For first-line treatment of unresectable locally advanced or metastatic urothelial carcinoma, cisplatin-containing chemotherapy regimens such as GC (gemcitabine + cisplatin) and MVAC (methotrexate + vincristine + adriamycin + cisplatin) are preferred if cisplatin can be tolerated. For patients who cannot tolerate cisplatin, carboplatin-containing regimens, such as the gemcitabine + carboplatin regimen, can be considered. For other patients, chemotherapy agents such as PD-1 inhibitors, such as pabulizumab or atezolizumab, gemcitabine + paclitaxel, or gemcitabine alone can be used. PD-1 inhibitors are preferred for patients with PD-L1 expression. Second-line treatment options currently include immunotherapy, targeted therapy, ADC therapy, and chemotherapy. There is currently no standard treatment for third-line and later treatments, and appropriate chemotherapy, targeted therapy, and biologics are required depending on the availability of frontline drugs.

[0004] In summary, there are no effective follow-up treatments for patients who have failed chemotherapy regimens containing platinum agents or PD-1 inhibitors, and new drugs need to be explored.

[0005] Breast cancer ranks first in incidence and fifth in mortality among malignant tumors in women in China, posing a serious threat to the health of Chinese women (Non-Patent Document 2). In 2015, there were approximately 304,000 new cases of breast cancer in China, with an incidence rate of 45.29 per 100,000 and more than 70,000 deaths. It is said that approximately 3% to 10% of new cases of breast cancer have distant metastasis at the time of diagnosis, and 30% of early-stage cases develop into advanced breast cancer. The five-year survival rate for advanced breast cancer is only 20% (Non-Patent Document 3).

[0006] Chemotherapy is essential for inoperable locally advanced, recurrent, or metastatic breast cancer patients who are hormone receptor-negative, HER-2-positive, have a short postoperative disease-free survival (<2 years), have rapidly progressing tumors, have severe symptoms, have widespread visceral metastases, are hormone receptor-positive, or are refractory to endocrine therapy. For patients with chemotherapy-naive patients (including adjuvant chemotherapy), anthracycline- and / or paclitaxel-based regimens are recommended as first-line chemotherapy. For patients with metastatic breast cancer who have not responded to anthracycline therapy or are approaching cumulative dose limits and have not previously received paclitaxel, paclitaxel-based regimens or single-agent paclitaxel regimens are preferred as first-line chemotherapy. Patients who have received paclitaxel in adjuvant chemotherapy and have had recurrence for more than one year can receive paclitaxel again, with priority given to other drug options, including adjuvant chemotherapy and drugs not used during the treatment phase. In recurrent metastatic breast cancer where both anthracyclines and paclitaxel have failed neoadjuvant and adjuvant therapy, there is no standard chemotherapy regimen, and drugs to consider are capecitabine, vincristine, gemcitabine, platinum-based drugs, eribulin, utiderone, other classes of paclitaxel (e.g., albumin-paclitaxel), and liposomal doxorubicin, either as single or combined regimens, so the continued development and exploration of novel agents for backline treatment remains necessary.

[0007] Classical osteosarcoma is the most common primary malignant tumor of bone, with an annual incidence of approximately 2–3 cases per million, accounting for only 0.2% of human malignancies. Osteosarcoma can occur at any age, most commonly during adolescence; approximately 75% of patients are between 15–25 years of age, with a median age of 20 years. It is relatively rare in patients younger than 6 years or older than 60 years. The disease is more common in men than women by a ratio of approximately 1.4:1, with this difference being particularly pronounced in patients younger than 20 years. Approximately 20% of patients have established lung metastases at initial diagnosis, and an additional 30% develop lung metastases within one year of osteosarcoma diagnosis. Before the 1970s, treatment for osteosarcoma consisted solely of surgery, primarily amputation, and the average 5-year survival rate was only 19.7%. In the late 1970s, a regimen of neoadjuvant chemotherapy + surgery + adjuvant chemotherapy was developed, and the 5-year survival rate has improved significantly to 60-80% (Non-Patent Document 4), which remains the same to the present day. On the other hand, the survival rate of patients with metastatic or recurrent osteosarcoma has remained almost unchanged over the past 30 years, with the overall 5-year survival rate remaining at approximately 20% (Non-Patent Document 5).

[0008] There are many chemotherapy regimens for osteosarcoma worldwide, and although there is no particular unified regimen, their effectiveness is similar depending on the type of drug used and its dosage intensity. First-line chemotherapy for osteosarcoma includes doxorubicin, cisplatin, isocyclophosphamide, and high-dose methotrexate, and similar drugs are also used in neoadjuvant chemotherapy. Two or more drugs are used for each patient at sufficient dosage intensity. Compared with doublet therapy, triplet therapy had a 5-year EFS (time-free survival) rate of 58% and 48%, and a 5-year OS rate of 70% and 62%. For osteosarcoma patients who have failed first-line chemotherapy, second-line treatments that offer benefits to overall survival are not yet available, so participation in clinical trials offers an opportunity to obtain better treatment outcomes and new treatments.

[0009] In summary, in addition to surgery, chemotherapy has become an important tool in clinical management at present. Patients with advanced or unresectable disease who are ineffective in first-line chemotherapy have a poor prognosis, and the medical evidence for targeted therapy and immunotherapy in osteosarcoma is still insufficient. The exploration or adoption of new cytotoxic or targeted drug therapies will provide opportunities for second-line treatment.

[0010] Soft tissue sarcomas are a group of malignant tumors arising from non-epithelial extraskeletal tissues, accounting for approximately 0.8% of all human malignant tumors. The annual incidence rate is estimated to be approximately 3.4 per 100,000 in the United States, approximately 40-5 per 100,000 in Europe, and approximately 2.38 per 100,000 in China (Non-Patent Document 6). Patients develop sarcomas regardless of age, with a male-to-female ratio of nearly 1:1 in China. The incidence rate increases significantly with age; when corrected for age, an 80-year-old is approximately eight times that of a 30-year-old (Non-Patent Document 7). Soft tissue sarcomas primarily arise from the mesoderm, with some arising from the neuroectoderm. These tumors primarily comprise muscle, fat, fibrous tissue, blood vessels, and peripheral nerve tissue. The most common site of origin is the extremities (approximately 53%), followed by the retroperitoneum (19%), trunk (12%), and head and neck (11%). There are 12 major categories based on tissue origin, with over 50 subtypes based on distinct morphologies and biological behaviors. Common subtypes are undifferentiated pleomorphic sarcoma, liposarcoma, leiomyosarcoma, and synovial sarcoma. The most common soft tissue sarcoma in adolescents and children is rhabdomyosarcoma.

[0011] Soft tissue sarcomas are primarily painless tumors that gradually enlarge and are difficult to treat, lasting from months to years. As the tumor gradually enlarges, it can compress nerves and blood vessels, causing pain, numbness, and even edema in the hands and feet. In some cases, the tumor may grow rapidly over a short period of time, accompanied by elevated skin temperature and enlarged surrounding lymph nodes. Caution is required as the tumor's malignancy may increase. Soft tissue sarcomas are characterized by their highly malignant nature, short course, rapid onset of hematogenous metastasis, and a high likelihood of recurrence after treatment. The most common site of metastasis for extremity sarcomas is the lung, while retroperitoneal and gastrointestinal sarcomas most commonly metastasize to the liver. The overall 5-year survival rate for soft tissue sarcomas is approximately 60-80%.

[0012] Soft tissue sarcomas are treated with a comprehensive treatment strategy centered on surgery. Radiation therapy, chemotherapy, or concurrent radiation therapy is considered one of the most effective treatments. With the advancement of multimodal oncological treatment, neoadjuvant therapy plus surgery plus chemotherapy has gradually been introduced, offering benefits such as tumor shrinkage, increased likelihood of limb preservation, improved local control rates, reduced risk of distant metastasis and recurrence, and improved patient survival. Chemosensitivity is an important basis for selecting chemotherapy for soft tissue sarcoma. Anthracycline-based regimens are preferred for neoadjuvant and adjuvant chemotherapy. However, in patients with high-grade soft tissue sarcoma, 40–50% experience local recurrence and more than 50% experience distant metastasis even after surgery. For patients with metastatic or recurrent incompletely resected tumors, chemotherapy is palliative chemotherapy, which aims to reduce and stabilize the tumor, thereby alleviating symptoms, extending survival, and improving quality of life.

[0013] For metastatic or recurrent unresectable tumors, individualized palliative chemotherapy needs to be developed. In patients with metastatic non-polyblastic rhabdomyosarcoma, chemotherapy regimens should alternate between VAC / VI / VCD / IE depending on the high-risk group selected. On the other hand, doxorubicin and isocyclophosphamide are the backbone drugs for nonspecific soft tissue sarcoma. There are no approved second-line chemotherapy regimens for nonspecific soft tissue sarcoma.

[0014] In summary, participation in clinical trials is one of the recommended options for patients with soft tissue sarcoma, which is highly malignant and has failed first-line treatment, and second-line treatment options are limited in terms of drug type and efficacy. There is an urgent clinical need to improve the treatment of patients with soft tissue sarcoma and to explore new drugs.

[0015] Mitoxantrone hydrochloride is an anthraquinone chemotherapy agent approved by the FDA for the treatment of multiple sclerosis, prostate cancer, and acute myeloid leukemia, and is approved in China for patients with lymphoma, leukemia, breast cancer, and other conditions. The recommended dose is 12-14 mg / m2 as a single agent once every 3-4 weeks for adults. 2 , or 4–8 mg / m once daily for 3–5 days every 2–3 weeks2 When used in combination, the dose is 5-10 mg / m 2 Mitoxantrone is an anthracycline drug whose main side effects are cardiac toxicity, bone marrow suppression, and gastrointestinal reactions. While bone marrow suppression and gastrointestinal reactions can be resolved with appropriate drug administration, cardiac toxicity is often severe and is the most serious side effect of anthracyclines. Clinical studies and practical observations have shown that most anthracycline-induced cardiac toxicity is progressive and irreversible, especially in patients receiving an anthracycline for the first time. Chronic dose-limiting cumulative toxicity—cardiotoxicity—is a common concern for clinicians. Therefore, although approved for this indication, mitoxantrone is not recommended for breast cancer treatment in authoritative clinical dosing guidelines. Mitoxantrone has never been approved for urothelial carcinoma or bone and soft tissue sarcoma.

[0016] Drug sensitivity varies among tumors. Existing research has shown that the same drug may be administered differently for different indications. For example, Doxil (doxorubicin hydrochloride liposomal) is approved by the FDA for three indications: (1) ovarian cancer, with a recommended dose of 50 mg / m 2 , intravenously administered every 4 weeks; (2) Kaposi's sarcoma, recommended dose 20 mg / m 2 , intravenously administered every 3 weeks, (3) multiple myeloma, recommended dose 30 mg / m 2 , administered intravenously on day 4 after bortezomib administration. Abraxane (injectable paclitaxel [albumin-bound]) is also approved by the FDA for the following three indications: (1) metastatic breast cancer: recommended dose 260 mg / m 2 , administered intravenously over 30 minutes every 3 weeks; (2) Non-small cell lung cancer: Recommended dose 100 mg / m 2 , administered intravenously over 30 minutes on days 1, 8, and 15 every 21 days; carboplatin is administered immediately after paclitaxel (albumin-bound) on day 1 and every 21 days; (3) Pancreatic cancer: recommended dose 125 mg / m 2It is administered intravenously over 30 to 40 minutes once on days 1, 8, and 15 of a 28-day cycle, and gemcitabine is administered immediately after paclitaxel (albumin-bound) injection. Furthermore, for example, the recommended starting dose for AmBisome (injectable liposomal amphotericin B) for the following indications is: (1) empirical treatment: recommended dose 3 mg / kg / day; (2) systemic fungal infections (Aspergillus, Candida, Cryptococcus): recommended dose 3-5 mg / kg / day; (3) cryptococcal meningitis in HIV-infected patients: recommended dose 6 mg / kg / day (days 1-5) and 3 mg / kg / day (days 4 and 21); (4) immunocompromised patients with visceral leishmaniasis: recommended dose 4 mg / kg / day (days 1-5) and 4 mg / kg / day (days 10, 17, 24, 31, and 38). Dosage and administration data are determined individually based on the patient's specific needs to maximize efficacy and minimize toxicity or adverse effects. It is clear that safe and effective dosages vary depending on the indication, even for the same drug. To achieve maximum efficacy and minimal toxicity or side effects, and to realize safe and effective treatment, dosage and administration data must be tailored to the specific disease and patient conditions. Mitoxantrone liposome is a special dosage form that differs from regular injections, and its absorption, distribution, and metabolism after administration are very complex, making it difficult to simply extrapolate to different indications, especially to the treatment of different tumors.

[0017] Therefore, it is necessary to systematically investigate whether mitoxantrone liposome is suitable for the treatment of urothelial carcinoma, breast cancer, and bone and soft tissue sarcoma, clarify its safety and effective dose, and provide reference for clinical treatment. [Prior art documents] [Non-patent literature]

[0018] [Non-Patent Document 1] Freddie, Bray, Jacques, Ferlay, Isabelle, & Soerjomataram, etal. (2018). Global cancer statistics 2018: globocan estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA: a cancer journal for monks. [Non-patent document 2] Zheng Rongshou, Sun Kexin, Zhang S. et al. Analysis of the prevalence of malignant tumors in China in 2015. Chinese Journal of Oncology Vol.41, No.1, January 2019 [Non-patent document 3] Breast Cancer Specialist Committee of the National Cancer Research Center of China, Standard Treatment Guidelines for Advanced Breast Cancer in China (2020 Edition) [Non-patent document 4] Niu Xiaohui. Current status and prospects of comprehensive treatment of osteosarcoma in China [J]. Chinese Journal of Anatomy and Clinics, 2019(01):1-5. [Non-Patent Document 5] Meyers PA, Healey JH, Chou AJ, et al. Addition of pamidronate to chemotherapy for the treatment of osteosarcoma[J].Cancer,2011,117(8):1736-1744. [Non-patent document 6] Siegel RL, Miller KD, Fuchs HE, et al. Cancer Statistics,2021[J].CA:A Cancer Journal for Clinicians,2021,71(1). [Non-Patent Document 7] Burningham Z, Hashibe M, Spector L, et al. The epidemiology of sarcoma[J].Clin Sarcoma Res,2012,2(1):14 Summary of the Invention

[0019] The present invention provides the use of mitoxantrone hydrochloride liposomes in the manufacture of a medicament for treating urothelial cancer, breast cancer, and bone and soft tissue sarcoma.

[0020] Preferably, the urothelial carcinoma is locally advanced or metastatic urothelial carcinoma, more preferably, the urothelial carcinoma is locally advanced or metastatic urothelial carcinoma that has failed treatment with a platinum-containing chemotherapy regimen and / or a PD-1 inhibitor; or locally advanced and metastatic urothelial carcinoma that has failed treatment with a platinum-containing chemotherapy regimen but refused treatment with a PD-1 inhibitor; or locally advanced or metastatic urothelial carcinoma that is intolerant to cisplatin.

[0021] Preferably, the breast cancer is HER-2-negative breast cancer, more preferably, the breast cancer is locally advanced or recurrent / metastatic HER-2-negative breast cancer; or hormone receptor-negative HER-2-negative breast cancer, or hormone receptor-positive HER-2-negative breast cancer that is not suitable for or resistant to endocrine therapy; or HER-2-negative breast cancer that has failed treatment with anthracycline and / or paclitaxel drugs; preferably, the HER-2-negative breast cancer includes breast cancer that is confirmed to be immunohistochemically HER-2 0 or 1+, or immunohistochemically HER-2 2+ but negative by in situ hybridization.

[0022] Preferably, the bone and soft tissue sarcoma is an advanced bone and soft tissue sarcoma, more preferably a metastatic or locally advanced bone and soft tissue sarcoma that has failed at least first line treatment.

[0023] Preferably, mitoxantrone hydrochloride liposome is used as the sole active ingredient in the manufacture of a medicament for treating urothelial cancer, breast cancer, and bone and soft tissue sarcoma.

[0024] Preferably, the drug is in an injectable dosage form, including liquid injection, injectable powder, injectable tablet, etc. Preferably, the drug is a liquid injection.

[0025] According to an embodiment of the present invention, when the drug is a liquid injection, the drug contains 0.5 to 5 mg / ml, preferably 1 to 2 mg / ml, more preferably 1 mg / ml, in terms of mitoxantrone.

[0026] The present invention also provides a method for treating urothelial carcinoma, breast cancer, and bone and soft tissue sarcoma, the method comprising administering a therapeutically effective amount of mitoxantrone hydrochloride liposome to a patient in need of treatment.

[0027] The present invention further provides mitoxantrone hydrochloride liposomes for the treatment of urothelial cancer, breast cancer, and bone and soft tissue sarcoma.

[0028] Preferably, the urothelial carcinoma is locally advanced or metastatic urothelial carcinoma, more preferably, the urothelial carcinoma is locally advanced or metastatic urothelial carcinoma that has failed treatment with a platinum-containing chemotherapy regimen and / or a PD-1 inhibitor; or locally advanced and metastatic urothelial carcinoma that has failed treatment with a platinum-containing chemotherapy regimen but refused treatment with a PD-1 inhibitor; or locally advanced or metastatic urothelial carcinoma that is intolerant to cisplatin.

[0029] Preferably, the breast cancer is HER-2-negative breast cancer, more preferably, the breast cancer is locally advanced or recurrent / metastatic HER-2-negative breast cancer; or hormone receptor-negative HER-2-negative breast cancer, or hormone receptor-positive HER-2-negative breast cancer that is not suitable for or resistant to endocrine therapy; or HER-2-negative breast cancer that has failed treatment with anthracycline and / or paclitaxel drugs; preferably, the HER-2-negative breast cancer includes breast cancer that is confirmed to be immunohistochemically HER-2 0 or 1+, or immunohistochemically HER-2 2+ but negative by in situ hybridization.

[0030] Preferably, the bone and soft tissue sarcoma is an advanced bone and soft tissue sarcoma, more preferably a metastatic or locally advanced bone and soft tissue sarcoma that has failed at least first line treatment.

[0031] Preferably, the therapeutically effective amount is a mitoxantrone dose of 8 to 30 mg / m 2 , more preferably 12 to 20 mg / m 2 or 16 to 30 mg / m 2 Specifically, for example, 12 mg / m 2 , 14 mg / m 2 , 16 mg / m 2 , 18 mg / m 2 , 20 mg / m 2 is.

[0032] Preferably, the administration method according to the present invention is intravenous administration. Preferably, the administration cycle is every 3 weeks. Preferably, the treatment given to the patient is 6 to 8 cycles. Preferably, the infusion administration time of the liposome pharmaceutical formulation per intravenous administration is 30 to 120 minutes, preferably 60 to 120 minutes, more preferably 60±15 minutes.

[0033] The present invention further provides mitoxantrone hydrochloride liposomes for treating urothelial cancer, breast cancer, and bone and soft tissue sarcoma in a patient.

[0034] In some embodiments, the liposomes are in the form of an injectable preparation, including a liquid injectable preparation, an injectable powder, an injectable tablet, etc. When the drug is a liquid injectable preparation, the drug contains 0.5 to 5 mg / ml, preferably 1 to 2 mg / ml, more preferably 1 mg / ml, of mitoxantrone equivalent.

[0035] In some embodiments, the liposomes are used alone to treat urothelial carcinoma, breast cancer, bone and soft tissue sarcoma in a patient.

[0036] In some embodiments, the therapeutically effective amount of the liposome is 8 to 30 mg / m mitoxantrone equivalent. 2 , more preferably 12 to 20 mg / m 2 or 16 to 30 mg / m 2 Specifically, for example, 12 mg / m 2 , 14 mg / m 2 , 16 mg / m 2 , 18 mg / m 2 , 20 mg / m 2 is.

[0037] In some embodiments, the liposome is administered intravenously. Preferably, the administration cycle is every 3 weeks. Preferably, the patient is administered 6 to 8 cycles of treatment. Preferably, the liposome is administered intravenously for 30 to 120 minutes, preferably 60 to 120 minutes, more preferably 60±15 minutes, per intravenous administration.

[0038] In the context of the present invention, the doses stated are stated in terms of mitoxantrone unless otherwise specified.

[0039] In the context of the present invention, the mitoxantrone hydrochloride liposomes may be prepared by any method known in the art, and may be mitoxantrone hydrochloride liposomes prepared by any method disclosed in the prior art, for example, the method disclosed in WO2008 / 080367A1.

[0040] In some embodiments, the drug or mitoxantrone hydrochloride liposomes have one or more of the following properties:

[0041] (i) the mitoxantrone hydrochloride liposome has a particle size of about 30 to 80 nm, for example, about 35 to 75 nm, about 40 to 70 nm, about 40 to 60 nm, or about 60 nm;

[0042] (ii) mitoxantrone hydrochloride forms an insoluble precipitate with multivalent counterions (e.g., sulfate, citrate, or phosphate) within the liposome;

[0043] (iii) the phospholipid bilayer in the mitoxantrone hydrochloride liposome comprises a phospholipid having a phase transition temperature (Tm) greater than body temperature, such that the liposome has a phase transition temperature greater than body temperature, for example, the phospholipid is selected from hydrogenated soybean lecithin, phosphatidylcholine, hydrogenated egg yolk lecithin, lecithin diphosphatidate, lecithin distearate, or any combination thereof;

[0044] (iv) The phospholipid bilayer in the mitoxantrone hydrochloride liposome contains hydrogenated soybean lecithin, cholesterol, and distearoylphosphatidylethanolamine modified with polyethylene glycol 2000 (DSPE-PEG2000).

[0045] (iiv) The phospholipid bilayer membrane in the mitoxantrone hydrochloride liposome contains hydrogenated soybean lecithin, cholesterol, and distearoylphosphatidylethanolamine modified with polyethylene glycol 2000 in a mass ratio of approximately 3:1:1, and mitoxantrone hydrochloride forms an insoluble precipitate with polyvalent acid ions in the liposome. The particle size of the mitoxantrone hydrochloride liposome is approximately 60 nm.

[0046] (iiiv) Mitoxantrone hydrochloride liposome is mitoxantrone hydrochloride liposome of China National Drug Registration H20220001.

[0047] In the context of the present invention, the mitoxantrone hydrochloride liposomes have a particle size of approximately 30 to 80 nm and comprise 1) the active ingredient mitoxantrone, which forms an insoluble precipitate with polyvalent counterions within the liposome, and 2) a phospholipid bilayer containing a phospholipid having a phase transition temperature (Tm) higher than body temperature, such that the liposomes have a Tm higher than body temperature. The phospholipid having a Tm higher than body temperature is phosphatidylcholine, hydrogenated soybean lecithin, hydrogenated egg yolk lecithin, diphosphatidic acid lecithin, distearic acid lecithin, or any combination thereof. The particle size is approximately 35 to 75 nm, preferably 40 to 70 nm, more preferably 40 to 60 nm, and particularly preferably 60 nm. Preferably, the phospholipid bilayer contains hydrogenated soybean lecithin, cholesterol, and distearoylphosphatidylethanolamine modified with polyethylene glycol 2000 in a mass ratio of 3:1:1, the particle size is about 60 nm, and the counter ion is sulfate. Preferably, the phospholipid bilayer of the liposome contains hydrogenated soybean lecithin, cholesterol, and distearoylphosphatidylethanolamine modified with polyethylene glycol 2000 in a mass ratio of 3:1:1, the particle size is about 40 to 60 nm, the counter ion is sulfate, and the weight ratio of HSPC:Chol:DSPE-PEG2000:mitoxantrone in the liposome is 9.58:3.19:3.19:1.

[0048] In the context of the present invention, mitoxantrone liposomes are prepared as follows: HSPC (hydrogenated soybean lecithin), Chol (cholesterol), and DSPE-PEG2000 (distearoylphosphatidylethanolamine modified with polyethylene glycol 2000) are weighed in a mass ratio of 3:1:1 and dissolved in 95% ethanol to obtain a clear solution. 300 mM ammonium sulfate solution is added to this ethanol solution of phospholipids, and the mixture is hydrated by shaking at 60-65°C for 1 hour to obtain heterogeneous multicompartment liposomes. The liposomes are then reduced in size using a microjet device. The resulting sample is diluted 200-fold with 0.9% NaCl solution and measured using a NanoZS. The average particle size is approximately 60 nm, with the main peak located between 40 and 60 nm. Next, the external phase of the blank liposomes is removed using an ultrafiltration device to remove ammonium sulfate and replaced with 290 mM sucrose and 10 mM glycine to form a transmembrane ammonium sulfate gradient. A solution of mitoxantrone hydrochloride (10 mg / mL) is added to the blank liposomes at a lipid-drug ratio of 16:1 and loaded at 60-65 °C. After approximately 1 hour of incubation, gel exclusion chromatography is used to demonstrate approximately 100% encapsulation efficiency. The weight ratio of HSPC:Chol:DSPE-PEG2000:mitoxantrone is 9.58:3.19:3.19:1, and the osmolality of the sucrose-glycine solution is close to physiological values.

[0049] It should be understood that the technical details and parameters in the exemplary manufacturing method described above can be adjusted and determined within reasonable limits by those skilled in the art. For example, the types of amino acids that can be substituted for glycine in the external phase used to form the transmembrane ammonium sulfate gradient include, but are not limited to, histidine, asparagine, glutamic acid, leucine, proline, and alanine. Again, for example, the mass ratios of HSPC, Chol, and DSPE-PEG2000 can be suitably adjusted. For example, in determining the lipid-drug ratio parameters for a particular liposomal pharmaceutical formulation, one skilled in the art can design, test, and ultimately arrive at an appropriate lipid-drug ratio that maximizes drug delivery while minimizing drug leakage. For the present mitoxantrone hydrochloride liposomal formulations, a wide range of lipid-drug ratios can be used, for example, as low as 2:1 or as high as 30:1, 40:1, or 50:1. A more preferred lipid-drug ratio may be about (15-20):1, for example, about 15:1, 16:1, 17:1, 18:1, 19:1, or 20:1. Therefore, several desirable properties of the above-described mitoxantrone hydrochloride liposomal formulations are important, and the methodologies for achieving these properties are diverse.

[0050] The first-line, second-line or higher-line drugs for the treatment of urothelial carcinoma, breast cancer, and bone and soft tissue sarcoma according to the present invention refer to first-line, second-line or higher-line drugs approved by drug regulatory authorities in China or overseas (e.g., the United States, the EU, Japan, South Korea, etc.) for the treatment of urothelial carcinoma, breast cancer, and bone and soft tissue sarcoma, including, but not limited to, FDA-approved paclitaxel drugs, PD-1 inhibitors, isocyclophosphamide, etc.

[0051] Beneficial effects In the research conducted by the applicant of the present invention, it was unexpectedly found that mitoxantrone hydrochloride liposome has a better therapeutic effect on urothelial cancer, breast cancer, and bone and soft tissue sarcoma, and can be used to manufacture related medicines. DETAILED DESCRIPTION OF THE INVENTION

[0052] The technical solutions of the present invention are described in further detail below in connection with specific embodiments. It is understood that the following embodiments are only for illustrative purposes and should not be construed as limiting the protection scope of the present invention. Any technology implemented based on the above elements of the present invention will fall within the protection scope intended by the present invention.

[0053] Unless otherwise specified, the components and reagents used in the following embodiments are commercially available products or can be prepared by known methods.

[0054] In the present invention, the following abbreviations are used: CR: Complete response as defined by the internationally recognized Research Evaluation Criteria for the Outcome of Solid Tumors (RECIST 1.1). PR: Partial response as defined by the internationally recognized Research Evaluation Criteria for the Outcome of Solid Tumors (RECIST 1.1). SD: Stable disease as defined by the internationally recognized Research Evaluation Criteria for the Outcome of Solid Tumors (RECIST 1.1). PD: Progressive disease as defined by the internationally recognized Research Evaluation Criteria for the Outcome of Solid Tumors (RECIST 1.1). Objective remission rate (ORR) = (CR + PR) / total evaluable cases × 100%. Disease control rate (DCR) = (CR + PR + SD) / total evaluable cases × 100%.

[0055] The present inventors conducted clinical trials to investigate the effectiveness of mitoxantrone hydrochloride liposomes in the treatment of urothelial carcinoma, breast cancer, and bone and soft tissue sarcoma. Compared with standard mitoxantrone hydrochloride injections, mitoxantrone hydrochloride liposomes were found to be more effective in treating urothelial carcinoma, breast cancer, and bone and soft tissue sarcoma with fewer side effects. The mitoxantrone hydrochloride liposome injections used in the following examples were provided by CSPC Group Zhongnuo Pharmaceutical (Shijiazhuang) Co., Ltd. (National Pharmaceutical Association No. H20220001). [Example]

[0056] Example 1: Phase II Clinical Trial to Evaluate the Efficacy and Safety of Mitoxantrone Hydrochloride Liposomal Injection for Unresectable Locally Advanced or Metastatic Urothelial Carcinoma This is an open, multicenter, phase II clinical trial to evaluate the efficacy and safety of mitoxantrone hydrochloride liposome injection in enrolled subjects for the treatment of unresectable locally advanced or metastatic urothelial carcinoma.

[0057] 1. Clinical trial design 1. Clinical trial method Forty patients with unresectable locally advanced or metastatic urothelial carcinoma were enrolled. The study subjects were primarily those who had failed platinum-based chemotherapy regimens and PD-1 inhibitor treatment (including those not receiving PD-1 inhibitors), and those who could not tolerate cisplatin and were receiving systemic therapy. This study consisted of a screening period, a treatment period, and a follow-up period. The screening period was 28 days, and eligible subjects were screened to proceed to the treatment period. The treatment period consisted of mitoxantrone hydrochloride liposomal injection 20 mg / m 2The drug was administered as a single agent every 3 weeks (q3w) on the first day of each cycle for six cycles. A follow-up period was conducted after the completion of treatment. During the follow-up period, safety follow-up after the final dose (28 days after the final dose) and survival follow-up (every 6 weeks after the final dose) were performed. Safety assessments were performed from the first dose until 28 days after the final dose and before each cycle to determine whether to administer the next cycle. Tumor assessments were performed every two cycles during the study period according to RECIST 1.1 criteria until disease progression, death, or receipt of new antitumor therapy.

[0058] 2. Research Period This clinical trial was planned to last approximately 12 months per patient, including a 4-week (28-day) screening period, 6 cycles (18-week) of treatment, a final safety follow-up, 4 weeks (28 days) after the end of treatment, and survival follow-up every 6 weeks thereafter. The trial enrolled 40 subjects and lasted for a total of 18 to 24 months.

[0059] 2. Clinical trial group: Subjects who met all of the following inclusion criteria and none of the exclusion criteria were enrolled in this trial. (1) Selection criteria 1) Subjects voluntarily participate in the study and sign informed consent; 2) age ≥ 18 years, male or female; 3) histologically confirmed urothelial carcinoma;

[0060] 4) Subjects who have failed platinum-containing chemotherapy regimens and PD-1 inhibitor treatment (including subjects who have not received PD-1 inhibitors), or subjects who cannot tolerate cisplatin and are candidates for systemic therapy; 5) presence of at least one evaluable lesion meeting RECIST 1.1 criteria at baseline; 6) ECOG score is 0 to 1;

[0061] 7) Subjects' laboratory test results met the following requirements: Absolute neutrophil count (ANC) ≥ 1.5x10 9 / L (not receiving leukocytosis treatment with G-CSF within 1 week prior to the test); Hemoglobin (Hb) > 9.0 g / dL (no red blood cell transfusion within 1 week prior to the test); Platelets ≥ 75x10 9 / L (no platelet transfusion within one week prior to the test) Creatinine ≤ 1.5xULN; ·Total bilirubin ≤1.5xULN (≤3xULN for subjects with liver metastases); ·Alanine aminotransferase (AST) / aspartate aminotransferase (ALT) ≤3xULN (≤5xULN for subjects with liver metastases); · Coagulation function: prothrombin time (PT), international normalized ratio (INR) ≤ 1.5 x ULN (if receiving anticoagulant medication such as warfarin, an INR ≤ 3 is acceptable); 8) Subject and partner are using effective contraception (e.g., combined hormonal (including estrogen and progestin) with ovulation suppression, progestin contraception with ovulation suppression, intrauterine device, intrauterine hormone-releasing system, bilateral vasectomy, bilateral ligation, abstinence from sexual activity, etc.) throughout the study and for 6 months after the end of the final dose; female subjects are HCG-negative in urine or blood (excluding menopause and hysterectomy).

[0062] (2) Exclusion criteria 1) Severe hypersensitivity to mitoxantrone or liposomal drugs; 2) Subjects with brain or meningeal metastases; 3) history of allogeneic organ transplantation or allogeneic bone marrow transplantation; 4) survival time <12 weeks; 5) Subjects with active hepatitis B (HbsAg or HBcAb positive, HBV DNA ≥ 2000 IU / mL), active hepatitis C (HCV antibody positive, HCV RNA above the lower limit of the testing center test), or HIV antibody positive;

[0063] 6) Active bacterial or fungal infection or viral infection requiring intravenous infusion within 1 week prior to the first administration of this drug; 7) Subjects who have received anti-cancer drug treatment within 4 weeks prior to the first administration (within 2 weeks prior to administration of herbal medicines or herbal combination medicines); 8) Treatment with other investigational drugs within 4 weeks prior to the first dose; 9) Have undergone major surgery within 3 months prior to the first administration or are scheduled to undergo major surgery during the study period; 10) Presence of Grade 1 or higher antineoplastic drug pretreatment toxicity (excluding alopecia, pigmentation, or other toxicity deemed by the study to pose no safety risk to the subject);

[0064] 11) Serious thromboembolic events, such as cerebrovascular accident (including transient ischemic attack) or pulmonary embolism, within the past 6 months; 12) Other active malignant tumors within the past 3 years, excluding curable localized cancers such as basal cell or squamous cell skin cancer, primary prostate cancer, cervical cancer, or breast cancer;

[0065] 13) Have any of the following cardiac abnormalities: ·Long QTc syndrome or QTc interval > 480 ms; Complete left bundle branch block, second-degree or third-degree atrioventricular block (except after treatment with pacemaker implantation); · have severe, uncontrolled heart failure requiring medical treatment; · History of chronic congestive heart failure, NYHA class ≥ 3; ·Cardiac ejection fraction less than 50% within 6 months; CTCAE > grade 2 valvular heart disease; · Uncontrolled hypertension (defined as systolic blood pressure >150mmHg or diastolic blood pressure >100mmHg on multiple measurements while under pharmacological control); - Myocardial infarction, unstable angina, severe pericardial disease, acute ischemia, or electrocardiogram findings of severe conduction pathway abnormalities within 6 months prior to screening.

[0066] 14) Patients who have been treated with adriamycin or other anthracyclines and have received a cumulative adriamycin dose of 350 mg / m 2 If it exceeds this amount (anthracycline equivalent dose calculation: adriamycin 1 mg = epirubicin 2 mg = pirarubicin 2 mg = desosorbicin 0.5 mg = mitocoxanthone 0.45 mg, excluding adriamycin liposome); 15) Pregnant or breastfeeding women;

[0067] 16)Having a serious and / or uncontrolled disease that, in the opinion of the investigator, may affect the patient's participation in this study (including, but not limited to, ineffectively controlled diabetes, kidney disease requiring dialysis, severe liver disease, life-threatening autoimmune and bleeding disorders, substance dependence, neurological disorders, etc.); 17) Any other condition that the investigator deems inappropriate for participation.

[0068] (3) Withdrawal from Treatment and Study Termination Criteria Subjects must discontinue treatment with the experimental drug if any of the following occurs during the study: 1) the subject exhibited intolerable toxicity and the investigator determined that the risks of continuing treatment with the experimental drug outweighed the benefits; 2) Complications that occur during treatment do not warrant further treatment; 3) have progressive disease as assessed by imaging;

[0069] 4) clinically assessed disease progression, significant deviation from the regimen, or subject poor compliance such that the investigator determined that continued treatment with the experimental drug would not be beneficial; 5) The subject is pregnant 6) Death occurs; 7) Meet any of the termination criteria for this clinical trial.

[0070] Subjects who discontinue treatment will be required to continue follow-up according to the study protocol unless the reason for discontinuation is death or they meet the discontinuation criteria listed below. Subjects have the right to withdraw from the study at any time and for any reason. A subject will withdraw from the study if any of the following occurs: 1) Lost to follow-up; 2) A request from the subject or family member (if the subject is unable to make the decision themselves) to discontinue the trial; 3) Completion of the study; 4) Other

[0071] 3. Research results For subjects who have not responded to platinum-containing chemotherapy regimens and PD-1 inhibitor treatment (including subjects not receiving PD-1 inhibitors), as well as for subjects who have received systemic therapy due to intolerance to cisplatin, there is no effective follow-on treatment, and the search for new drugs is urgently needed. The inventors of the present invention have discovered that by using mitoxantrone liposomes, which have sustained release, targeting, reduced toxicity, and improved efficacy when administered intravenously, the liposomal formulation can be used at higher doses than conventional injections of mitoxantrone, and can achieve the desired therapeutic effect with reduced side effects and improved efficacy as a monotherapy.

[0072] In the present study, at least four subjects (those treated with two cycles and those treated with one cycle) showed SD2 (those treated with one cycle and those treated with two cycles), with a DCR of 50% (2 / 4). Mitoxantrone liposome demonstrated good efficacy in patients with urothelial carcinoma, especially in patients who had failed platinum-containing chemotherapy regimens and PD-1 inhibitors. In the two subjects who were evaluated as SD, continued administration achieved better efficacy.

[0073] This regimen completes the treatment regimen for urothelial carcinoma, lays the foundation for drug combinations that will have an impact on first-line and second-line treatment, changes the traditional treatment mode for urothelial carcinoma, and hopes to apply similar highly effective and less toxic chemotherapeutic agents in the field of advanced solid tumors.

[0074] Example 2: Phase II clinical trial of mitoxantrone hydrochloride liposome injection for HER-2-negative advanced breast cancer This is an open, multicenter, phase II clinical trial in which enrolled subjects will be administered mitoxantrone hydrochloride liposome injection to evaluate its efficacy and safety in HER-2-negative advanced breast cancer.

[0075] 1. Clinical trial design 1. Clinical trial method This study was divided into a screening period, a treatment period, and a follow-up period. The screening period was 28 days, and subjects who passed the screening period entered the treatment period. During the treatment period, subjects received mitoxantrone hydrochloride liposomal injection at 20 mg / m 2 The drug was administered in six 3-week cycles. At the end of the six cycles, the investigators and sponsors would decide whether to continue treatment based on the benefits and risks to the subject. Follow-up would include end-of-treatment (28 days after the last dose) and survival follow-up (every 6 weeks after end-of-treatment), with all subjects required to receive survival follow-up by phone or in-person until death.

[0076] Tumor assessment: Tumor assessment was performed according to RECIST 1.1 criteria every two cycles during treatment and every 6 weeks during follow-up until disease progression or initiation of new anti-tumor therapy. Safety assessments: Vital signs, physical examination, weight, ECOG fitness score, 12-lead electrocardiogram, echocardiogram, and laboratory tests were performed before and at the end of each cycle. Routine blood tests were performed on days 8 and 15 of each cycle.

[0077] 2. Research Period This clinical trial will be conducted for approximately 12 months per patient, with a 4-week (28-day) screening period, 6 cycles (18-week) of treatment, and survival follow-up at the end of treatment (28 days after the last dose) and every 6 weeks thereafter until the patient is lost to follow-up or dies. The trial enrolled 73 subjects and lasted approximately 36 months.

[0078] 2. Clinical trial group: Subjects who met all of the following inclusion criteria and none of the exclusion criteria were enrolled in this trial.

[0079] (1) Selection Criteria: Participants must meet all of the following criteria: 1) Subjects voluntarily participate in the study and sign informed consent; 2) Age 18 years or older but younger than 75 years (including age 18 years or older but younger than 75 years); 3) Histopathologically confirmed HER-2 negative breast cancer (immunohistochemical HER-2 0 or 1+, including breast cancers confirmed as HER-2 2+ immunohistochemically negative by in situ hybridization); 4) Inoperable locally advanced or recurrent / metastatic breast cancer that has progressed after first-line chemotherapy and continues to progress thereafter;

[0080] 5) Hormone receptor-negative, or hormone receptor-positive but unsuitable for endocrine therapy, or endocrine therapy-resistant breast cancer; 6) previously treated with at least one anthracycline and one paclitaxel (subjects with no anthracycline history due to high risk factors for cardiac toxicity were also eligible), and up to four chemotherapy regimens; 7) at least one measurable lesion at baseline (RECIST 1.1 criteria); 8) ECOG score of 0-2;

[0081] 9) Good organ function (no blood transfusions or growth factor supportive therapy within 2 weeks prior to first administration of study drug), including: Absolute neutrophil count (ANC) ≥ 1.5 × 10 9 / L; Hemoglobin (Hb) ≥ 90g / L; Platelets ≥ 90 × 10 9 / L; Creatinine ≤ 1.5 times the upper limit of normal; Total bilirubin ≤ 1.5 times the upper limit of normal; Alanine aminotransferase (AST) and aspartate aminotransferase (ALT) ≤ 3 times the upper limit of normal (≤ 5 times the upper limit of normal in patients with liver metastases);

[0082] 10) a negative pregnancy test result and a commitment by subjects of childbearing age to use effective contraception or abstinence from the start of the study for 6 months after the final dose; 11) Good compliance and cooperation with follow-up visits.

[0083] (2) Exclusion Criteria Subjects who meet any of the following criteria will be excluded from this study: 1) Severe hypersensitivity to mitoxantrone or liposomal drugs; 2) History of other malignant tumors (excluding primary cervical cancer, basal cell carcinoma of the skin, and squamous cell carcinoma) within the past 3 years; 3) Subjects with brain or meningeal metastases; 4) Subjects with active hepatitis B (HbsAg or HBcAb positive, HBV DNA ≥ 2000 IU / mL), active hepatitis C (HCV antibody positive, HCV RNA above the lower limit of the testing center test), or HIV antibody positive;

[0084] 5) expected survival time < 3 months; 6) The cumulative dose of previously administered anthracyclines converted to doxorubicin is 350 mg / m 2(Calculation of anthracycline equivalent amount: doxorubicin 1 mg = epirubicin 2 mg = pirizubicin 2 mg = erythromycin 2 mg = desosorbicin 0.5 mg = mitocoxantrone 0.45 mg, except for adriamycin liposome); 7) No recovery of toxicity from previous antineoplastic therapy to Grade 1 or less (excluding alopecia, pigmentation, and other toxicities determined by the study to pose no safety risk to the subject);

[0085] 8) Have any of the following cardiac abnormalities: ·Long QTc syndrome or QTc interval > 480 ms; Complete left bundle branch block, second-degree or third-degree atrioventricular block; · have severe, uncontrolled heart failure requiring medical treatment; ·A history of chronic congestive heart failure and NYHA class ≥ 3; ·Cardiac ejection fraction less than 50% within 6 months prior to screening; ·Having valvular heart disease of CTCAE ≥ grade 3; -Having myocardial infarction, unstable angina, severe ventricular arrhythmia, severe pericardial disease, acute ischemia, or electrocardiogram findings of severe conduction pathway abnormalities within 6 months prior to screening.

[0086] 9) uncontrolled hypertension (systolic blood pressure ≥ 160 mmHg or diastolic blood pressure ≥ 100 mmHg on multiple measurements under pharmacological control); 10) Active bacterial, fungal, or viral infection requiring intravenous treatment within 1 week prior to the first dose; 11) Have received anti-tumor treatment (including chemotherapy, radiotherapy, molecular targeted therapy, immunotherapy, and endocrine therapy) within 4 weeks prior to the first administration, have received immunomodulators as adjuvant therapy for malignant tumors within 2 weeks prior to the first administration, or have received original anti-tumor herbal medicines (excluding those in the symptom support / relief category) within 2 weeks prior to the first administration. 12) Other investigational drugs administered within 4 weeks prior to the first dose; 13) Subjects who have undergone major surgery within 12 weeks prior to the first administration or who are scheduled to undergo major surgery during the study period;

[0087] 14) Deep vein thrombosis or arterial embolism within the past 6 months (including but not limited to superior and inferior vena cava thrombosis, deep vein thrombosis of the lower extremities, and pulmonary embolism); 15) breastfeeding women; 16) Have a serious and / or uncontrolled disease that, in the opinion of the investigator, may affect your participation in this study (including, but not limited to, ineffectively controlled diabetes, kidney disease requiring dialysis, severe liver disease, life-threatening autoimmune and bleeding disorders, substance dependence, or neurological disorders); 17) Any other condition that the investigator deems inappropriate for participation.

[0088] (3) Withdrawal from Treatment and Study Termination Criteria Subjects must discontinue treatment with the experimental drug if any of the following occurs during the study: 1) the subject exhibited intolerable toxicity and the investigator determined that the risks of continuing treatment with the experimental drug outweighed the benefits; 2) have progressive disease as assessed by imaging; 3) clinically assessed disease progression, significant deviation from the regimen, or subject poor compliance such that the investigator determined that continued treatment with the experimental drug would not be beneficial; 4) The subject is pregnant 5) Death occurs; 6) Meet any of the termination criteria for this clinical trial.

[0089] Subjects who discontinue treatment will be required to continue follow-up according to the study protocol unless the reason for discontinuation is death or they meet one of the discontinuation criteria listed below. Subjects have the right to withdraw from the study at any time and for any reason. A subject will be withdrawn from the study if any of the following apply: 1) Lost to follow-up; 2) the subject withdrew informed consent or the subject or family requested discontinuation of the trial; 3) Completion of the study; 4)Others.

[0090] 3. Research results There is an urgent need to explore new treatments for advanced breast cancer that has failed or recurred after previous treatments with anthracyclines or paclitaxel. In the study of the present invention, of the 20 evaluable cases, at least 6 cases achieved PR (1 case after 7 cycles of treatment, 3 cases after 6 cycles of treatment, 1 case after 5 cycles of treatment, and 1 case after 4 cycles of treatment), and 8 cases achieved SD (2 cases after 6 cycles of treatment, 2 cases after 5 cycles of treatment, 1 case after 4 cycles of treatment, 1 case after 3 cycles of treatment, and 2 cases after 2 cycles of treatment). ORR was 30% (6 / 20), and DCR was 70% (14 / 20).

[0091] Mitoxantrone liposome has shown favorable effects in breast cancer patients, especially those with advanced breast cancer who have failed or relapsed after previous anthracycline and paclitaxel therapy. Further favorable effects were observed in 14 patients who were evaluated as having partial response and stable disease after continued administration of the drug.

[0092] In the inventor's previous clinical trial, mitoxantrone hydrochloride monotherapy for recurrent / metastatic breast cancer was administered to 30 patients in the test group and 30 patients in the control group. The test group received mitoxantrone hydrochloride liposome injection solution at 20 mg / m 2 The control group received mitoxantrone hydrochloride injection at 14 mg / m 2 The results were PR in 4 cases, SD in 11 cases, ORR of 13.3% (4 / 30), and DCR of 50% (15 / 30) in the study group, and PR in 2 cases, SD in 7 cases, ORR of 6.7% (2 / 30), and DCR of 30% (9 / 30) in the control group.

[0093] The inventors of the present invention have used mitoxantrone liposomes, which have sustained release, targeting, reduced toxicity and enhanced effects when the drug is administered intravenously, not only at higher doses than conventional injections but also as monotherapy, thereby enhancing efficacy and reducing the possibility of side effects.

[0094] Example 3: Phase Ib clinical trial of mitoxantrone hydrochloride liposome injection for advanced bone and soft tissue sarcoma refractory to at least first-line treatment. This clinical trial is an open, multicenter, Phase Ib study to evaluate the safety and efficacy of mitoxantrone hydrochloride liposome injection in enrolled subjects with metastatic or unresectable bone and soft tissue sarcoma that has failed at least first-line treatment.

[0095] 1. Clinical trial design 1. Clinical trial method The trial will include a screening period, a treatment period, and a follow-up period. The screening period was 28 days, and subjects who passed the screening entered the treatment period. Subjects in the treatment period received mitoxantrone hydrochloride liposomal injection 20 mg / m on day 1 of a 3-week treatment cycle (q3w). 2 Patients received a total of six cycles of treatment. For subjects who completed six cycles of treatment, the investigator, in consultation with the sponsor, decided whether to continue the drug, taking into account the risks and benefits of the subject. The follow-up period included a safety follow-up after the final dose (28 ± 7 days after the final dose) and a survival follow-up (every six weeks after the final dose).

[0096] Safety assessments were performed from the first dose until 28 days after the final dose and before each cycle to determine whether to administer the next cycle. Tumor assessments were performed based on the RESIST 1.1 criteria every two cycles during the treatment period and every 6 weeks during the follow-up period until disease progression, death, or the administration of new antitumor therapy.

[0097] 2. Research Period This clinical trial will have a screening period of 4 weeks (28 days), a treatment period of 6 cycles (18 weeks), a final safety follow-up 4 weeks (28 days) after the end of administration, and survival follow-up every 6 weeks thereafter, and is expected to last approximately 12 months per patient. More than 50 subjects were enrolled in the study, and the total duration of the study ranged from 24 to 36 months.

[0098] 2. Study group: Subjects who met all of the following inclusion criteria and did not meet any of the exclusion criteria were enrolled in this study. (1) Selection criteria Subjects must meet all of the following criteria: 1) Subjects voluntarily participate in the study and sign informed consent; 2) age ≥ 18 years, male or female; 3) Histologically confirmed osteosarcoma or nonspecific soft tissue sarcoma (excluding types of soft tissue sarcoma such as embryonal / alveolar rhabdomyosarcoma and Ewing's sarcoma, highly chemotherapy-sensitive sarcomas such as adenoid soft tissue sarcoma and extraskeletal myxoid chondrosarcoma, gastrointestinal mesenchymal tumors, and sarcomas requiring special management such as advanced fibromatosis);

[0099] 4) metastatic or unresectable osteosarcoma or soft tissue sarcoma that has failed at least first-line treatment; 5) presence of at least one evaluable lesion meeting RECIST 1.1 criteria at baseline; 6) ECOG score of 0 to 1;

[0100] 7) Subjects' laboratory test results met the following requirements: Absolute neutrophil count (ANC) ≥ 1.5x10 9 / L (no leukocytosis treatment with G-CSF within 1 week prior to the test); Hemoglobin (Hb) ≥ 110 g / L (no red blood cell transfusion within 1 week prior to the test); Platelets ≥ 100x10 9 / L (no platelet transfusion within one week prior to the test) Creatinine ≤ 1.5xULN; ·Total bilirubin ≤1.5xULN (≤3.0xULN for subjects with liver metastases); alanine aminotransferase (AST) / aspartate aminotransferase (ALT) ≤ 3.0 x ULN (≤ 5.0 x ULN for subjects with liver metastases); Coagulation function: prothrombin time (PT), international normalized ratio (INR) ≤ 1.5xULN (if taking anticoagulant medication such as warfarin, an INR ≤ 3 is acceptable);

[0101] 8) Female subjects had negative urine or blood HCG (excluding menopause and hysterectomy); 9) The subject and their partner agree to use effective contraception (hormonal contraception (including estrogen and progestin) combined with ovulation suppression, progestin contraception combined with ovulation suppression, intrauterine device, intrauterine hormone-releasing system, bilateral ligation, vasectomy, abstinence from sexual activity, etc.) during the study and for 6 months after the end of the final dose;

[0102] (2) Exclusion criteria Subjects with any of the following conditions will be excluded from this study: 1) Severe hypersensitivity to mitoxantrone or liposomal drugs; 2) Subjects with brain or meningeal metastases; 3) history of allogeneic organ transplantation or allogeneic bone marrow transplantation;

[0103] 4) expected survival <12 weeks; 5) Chronic hepatitis B (HbsAg or HBcAb positive, HBV DNA ≥ 1000 IU / mL), hepatitis C (HCV antibody positive, HCV RNA quantification above the lower limit of the testing center's test value), or HIV antibody positive; 6) Presence of Grade 1 or higher antineoplastic drug pretreatment toxicity (excluding alopecia, pigmentation, or other toxicity deemed by the study to pose no safety risk to the subject);

[0104] 7) Active bacterial, fungal, or viral infection requiring systemic treatment within 1 week prior to the first dose; 8) Subjects who have received anti-cancer drug treatment within 4 weeks prior to the first administration (within 2 weeks prior to administration of herbal medicines or herbal combination medicines); 9) Treatment with other investigational drugs within 4 weeks prior to the first dose;

[0105] 10) Severe thrombosis, such as pulmonary embolism, visceral embolism, or deep vein thrombosis, within the past 6 months; 11) Other active malignant tumors within the past 3 years, excluding curable localized cancers such as basal cell or squamous cell skin cancer, primary prostate cancer, cervical cancer, or breast cancer;

[0106] 12) Have any of the following cardiac abnormalities: ·Long QTc syndrome or QTc interval > 480 ms; Complete left bundle branch block, second-degree or third-degree atrioventricular block (except after treatment with pacemaker implantation); · have severe, uncontrolled heart failure requiring medical treatment; · History of chronic congestive heart failure, NYHA class ≥ 3; ·Cardiac ejection fraction less than 50% within 6 months; ·Having valvular heart disease of CTCAE ≥ grade 3; · have uncontrolled hypertension (defined as systolic blood pressure >160mmHg or diastolic blood pressure >100mmHg on multiple measurements while under pharmacological control); - Myocardial infarction, unstable angina, severe pericardial disease, acute ischemia, or electrocardiogram findings of severe conduction pathway abnormalities within 6 months prior to screening.

[0107] 13) Patients who have been treated with adriamycin or other anthracyclines and have received a cumulative adriamycin dose of 350 mg / m 2If the cumulative dose of adriamycin liposome exceeds 900 mg / m (anthracycline equivalent dose calculation: adriamycin 1 mg = epirubicin 2 mg = pirarubicin 2 mg = desosorbicin 0.5 mg = mitocoxanthone 0.45 mg), 2 When it exceeds 14) Pregnant or breastfeeding women; 15) Have a serious and / or uncontrolled medical condition that, in the opinion of the investigator, may affect participation in this study (including, but not limited to, ineffectively controlled diabetes, kidney disease requiring dialysis, severe liver disease, life-threatening autoimmune and bleeding disorders, substance dependence, neurological disorders, etc.); 16) Any other condition that the investigator deems inappropriate for participation.

[0108] (3) Withdrawal from Treatment and Study Termination Criteria Subjects must discontinue treatment with the experimental drug if any of the following occurs during the study: 1) the subject exhibited intolerable toxicity and the investigator determined that the risks of continuing treatment with the experimental drug outweighed the benefits; 2) Complications that occur during treatment do not warrant further treatment; 3) have progressive disease as assessed by imaging;

[0109] 4) clinically assessed disease progression, significant deviation from the regimen, or subject poor compliance such that the investigator determined that continued treatment with the experimental drug would not be beneficial; 5) The subject is pregnant 6) Death occurs; 7) Meet any of the termination criteria for this clinical trial.

[0110] All subjects who discontinue treatment will be required to continue follow-up according to the study protocol unless the reason for discontinuation is death or they meet the discontinuation criteria listed below. Subjects have the right to withdraw from the study at any time and for any reason. A subject will withdraw from the study if any of the following occurs: 1) Lost to follow-up; 2) A request from the subject or family member (if the subject is unable to make the decision themselves) to discontinue the trial; 3) Completion of the study; 4)Others.

[0111] 3. Research results In patients with metastatic bone and soft tissue sarcoma and patients for whom surgical cure is not possible, the efficacy of second-line treatments after standard first-line therapy is low, making treatment improvement a clinical challenge and necessitating the investigation of new treatments. Mitoxantrone liposome demonstrated a significant improvement in efficacy in patients with advanced soft tissue sarcoma at a dose of 12 mg / m in the Hill Climbing Study (HE071-01). 2 and 14 mg / m 2 The inventors of the present invention have used mitoxantrone liposomes, which have sustained release, targeted properties, reduced toxicity, and improved efficacy when administered intravenously, which not only allows for higher doses than conventional injections but also allows for use as monotherapy, improving efficacy and reducing the possibility of side effects.

[0112] According to the test of the present invention, of the three subjects with osteosarcoma (two cycles of treatment, one cycle of treatment) evaluated, one was evaluated as SD (one cycle of treatment), and the DCR was shown to be 33.3% (1 / 3).

[0113] In soft tissue sarcoma, of 12 cases (1 case treated with 6 cycles, 2 cases treated with 5 cycles, 2 cases treated with 4 cycles, 1 case treated with 3 cycles, and 6 cases treated with 2 cycles), 8 cases were evaluated as SD (1 case treated with 6 cycles, 2 cases treated with 5 cycles, 2 cases treated with 4 cycles, 1 case treated with 3 cycles, and 2 cases treated with 2 cycles, of which 3 cases had a reduction in tumor burden), and the DCR was 66.7% (8 / 12).

[0114] Mitoxantrone liposome has shown favorable effects in patients with osteosarcoma and soft tissue sarcoma. Nine subjects who were evaluated as having SD continued receiving the drug and achieved better results.

[0115] The above is an illustrative description of the embodiments of the technical solutions of the present invention. It should be understood that the protection scope of the present invention is not limited to the above-mentioned embodiments. Any modifications, equivalent replacements, improvements, etc. made by those skilled in the art within the spirit and principle of the present invention shall be included in the protection scope of the claims of this application.

[0116] CROSS-REFERENCE TO RELATED APPLICATIONS The present invention claims priority to a prior application entitled "Applications of Mitoxantrone Hydrochloride Liposomes," filed with the State Intellectual Property Office of China under patent application number 202110410490.2 on April 16, 2021, the entire contents of which are incorporated herein by reference. This patent application cites PCT application WO2008 / 080367A1, filed on December 29, 2007, the entire disclosure of which is incorporated herein by reference.

Claims

1. Use of mitoxantrone hydrochloride liposomes in the manufacture of a medicament for treating urothelial cancer, breast cancer, and bone and soft tissue sarcoma.

2. Use of mitoxantrone hydrochloride liposomes as the sole active ingredient in the manufacture of a medicament for treating urothelial cancer, breast cancer, and bone and soft tissue sarcoma.

3. the urothelial carcinoma is locally advanced or metastatic urothelial carcinoma, more preferably the urothelial carcinoma is locally advanced or metastatic urothelial carcinoma that has failed treatment with a platinum-containing chemotherapy regimen and / or a PD-1 inhibitor; or locally advanced and metastatic urothelial carcinoma that has failed treatment with a platinum-containing chemotherapy regimen but refused treatment with a PD-1 inhibitor; or locally advanced or metastatic urothelial carcinoma that is intolerant to cisplatin; Preferably, the breast cancer is HER-2 negative breast cancer, more preferably, the breast cancer is locally advanced or recurrent / metastatic HER-2 negative breast cancer; or hormone receptor negative HER-2 negative breast cancer, or hormone receptor positive HER-2 negative breast cancer that is not suitable for or resistant to endocrine therapy; or HER-2 negative breast cancer that has failed treatment with anthracycline and / or paclitaxel drugs; preferably, the HER-2 negative breast cancer includes breast cancer that is confirmed to be immunohistochemically HER-2 0 or 1+, or immunohistochemically HER-2 2+ but negative by in situ hybridization, Preferably, the bone and soft tissue sarcoma is an advanced bone and soft tissue sarcoma, more preferably a metastatic or locally advanced bone and soft tissue sarcoma that has failed at least first-line treatment.

3. Use according to claim 1 or 2.

4. The drug is in an injection form, including liquid injections, injection powders, injection tablets, etc.; Preferably, the drug is a liquid injection, Preferably, the drug contains 0.5 to 5 mg / ml, preferably 1 to 2 mg / ml, more preferably 1 mg / ml, of mitoxantrone.

3. Use according to claim 1 or 2.

5. 1. A method for treating urothelial carcinoma, said method comprising administering to a patient in need thereof a therapeutically effective amount of mitoxantrone hydrochloride liposome; Preferably, the urothelial carcinoma is locally advanced or metastatic urothelial carcinoma, more preferably, the urothelial carcinoma is locally advanced or metastatic urothelial carcinoma that has failed treatment with a platinum-containing chemotherapy regimen and / or a PD-1 inhibitor; or locally advanced and metastatic urothelial carcinoma that has failed treatment with a platinum-containing chemotherapy regimen but refused treatment with a PD-1 inhibitor; or locally advanced or metastatic urothelial carcinoma that is intolerant to cisplatin.

6. 1. A method of treating breast cancer, said method comprising administering to a patient in need thereof a therapeutically effective amount of mitoxantrone hydrochloride liposome; Preferably, the breast cancer is HER-2 negative breast cancer, more preferably, the breast cancer is locally advanced or recurrent / metastatic HER-2 negative breast cancer; or hormone receptor negative HER-2 negative breast cancer, or hormone receptor positive HER-2 negative breast cancer that is not suitable for endocrine therapy or is resistant to endocrine therapy; or HER-2 negative breast cancer that has failed treatment with anthracycline and / or paclitaxel drugs; and preferably, the HER-2 negative breast cancer includes breast cancer that is confirmed to be immunohistochemically HER-2 0 or 1+ or immunohistochemically HER-2 2+ but negative by in situ hybridization.

7. 1. A method for treating bone and soft tissue sarcoma, said method comprising administering to a patient in need thereof a therapeutically effective amount of mitoxantrone hydrochloride liposome; Preferably, the bone and soft tissue sarcoma is advanced bone and soft tissue sarcoma, more preferably metastatic or locally advanced bone and soft tissue sarcoma that has failed at least first line therapy.

8. the administration route is intravenous administration, Preferably, the infusion time of the liposome pharmaceutical preparation is 30 to 120 minutes, preferably 60 to 120 minutes, more preferably 60±15 minutes per intravenous administration; Preferably, the administration cycle is every 4 weeks or every 3 weeks, preferably every 3 weeks; Preferably, the therapeutically effective amount is 8 to 30 mg / m2 in terms of mitoxantrone. ２ , more preferably 12 to 20 mg / m ２ or 16 to 30 mg / m ２ and more preferably 20 mg / m ２ That is, The method according to any one of claims 5 to 7.

9. Mitoxantrone hydrochloride liposome for treating urothelial cancer, breast cancer, and bone and soft tissue sarcoma, A therapeutically effective amount of mitoxantrone hydrochloride liposome is administered to a patient in need of treatment, and the therapeutically effective amount is 8 to 30 mg / m2 in terms of mitoxantrone. ２ , more preferably 12 to 20 mg / m ２ or 16 to 30 mg / m ２ and more preferably 20 mg / m ２ and Preferably, the administration method is intravenous administration, and the infusion administration time of the liposome pharmaceutical preparation is preferably 30 to 120 minutes, preferably 60 to 120 minutes, more preferably 60±15 minutes per intravenous administration; Preferably, the administration cycle is every 4 weeks or every 3 weeks, preferably every 3 weeks.

1. A mitoxantrone hydrochloride liposome comprising:

10. The mitoxantrone hydrochloride liposomes have one or more of the following properties: (i) the mitoxantrone hydrochloride liposomes have a particle size of about 30-80 nm, e.g., about 35-75 nm, about 40-70 nm, about 40-60 nm, or about 60 nm; (ii) mitoxantrone hydrochloride forms an insoluble precipitate with multivalent counterions (e.g., sulfate, citrate, or phosphate) within the liposome; (iii) the phospholipid bilayer in the mitoxantrone hydrochloride liposome comprises a phospholipid having a phase transition temperature (Tm) greater than body temperature, such that the liposome has a phase transition temperature (Tm) greater than body temperature, for example, the phospholipid is selected from hydrogenated soybean lecithin, phosphatidylcholine, hydrogenated egg yolk lecithin, lecithin diphosphatidate, lecithin distearate, or any combination thereof; (iv) the phospholipid bilayer in the mitoxantrone hydrochloride liposome comprises hydrogenated soybean lecithin, cholesterol, and distearoylphosphatidylethanolamine modified with polyethylene glycol 2000 (DSPE-PEG2000); (iii) the phospholipid bilayer membrane in the mitoxantrone hydrochloride liposome contains hydrogenated soybean lecithin, cholesterol, and distearoylphosphatidylethanolamine modified with polyethylene glycol 2000 in a mass ratio of about 3:1:1, mitoxantrone hydrochloride forms an insoluble precipitate with polyvalent acid ions in the liposome, and the particle size of the drug in the mitoxantrone hydrochloride liposome is about 60 nm; Preferably, the mitoxantrone hydrochloride liposome has a particle size of about 30 to 80 nm and comprises: 1) the active ingredient mitoxantrone, which forms an insoluble precipitate with polyvalent counterions within the liposome; and 2) a phospholipid bilayer comprising a phospholipid having a phase transition temperature (Tm) higher than body temperature, such that the liposome has a Tm higher than body temperature, wherein the phospholipid having a Tm higher than body temperature is phosphatidylcholine, hydrogenated soybean lecithin, hydrogenated egg yolk lecithin, diphosphatidic acid lecithin, distearic acid lecithin, or any combination thereof. Preferably, the particle size is about 35 to 75 nm, preferably 40 to 70 nm, more preferably 40 to 60 nm, and particularly preferably 60 nm; Alternatively, preferably, the phospholipid bilayer contains hydrogenated soybean lecithin, cholesterol, and distearoylphosphatidylethanolamine modified with polyethylene glycol 2000 in a mass ratio of 3:1:1, the particle size is about 60 nm, and the counter ion is sulfate ion; Alternatively, preferably, the phospholipid bilayer of the liposome contains hydrogenated soybean lecithin, cholesterol, and distearoylphosphatidylethanolamine modified with polyethylene glycol 2000 in a mass ratio of 3:1:1, the particle size is about 40 to 60 nm, the counter ion is a sulfate ion, and the weight ratio of HSPC:chol:DSPE-PEG2000:mitoxantrone in the liposome is 9.58:3.19:3.19:

1. The use, method or liposome according to any one of claims 1 to 9.