HK40082294A - Uses of mitoxantrone hydrochloride liposome - Google Patents

Description

Application of mitoxantrone hydrochloride liposome

The present invention claims priority to a prior application entitled "use of mitoxantrone hydrochloride liposomes" filed on 16.4.2021 by the intellectual property office of the chinese country with patent application number 202110410490.2. The entire disclosure of this prior application is incorporated herein by reference.

This patent application refers to PCT application WO2008/080367A1, filed on 12, 29 of 2007, the disclosure of which is incorporated by reference in its entirety.

Technical Field

The invention belongs to the field of anti-tumor, and particularly relates to application of mitoxantrone hydrochloride liposome, such as application in preparing medicines for treating urothelial carcinoma, breast cancer, bone and soft tissue sarcoma.

Background

Urothelial cancer is a multiple malignant tumor originating from urothelium, including renal pelvis cancer, ureter cancer, bladder cancer, urethral cancer and the like, and is the most common tumor of the urinary system. Globally, the incidence of bladder-derived tumors in 2018 ranks 12 in all solid tumors, accounting for 2.1% of total tumor deaths ("Freddie, bray, jacques, ferlay, isabelle, & sorjomataram, et al (2018). Global cancer stability 2018. The incidence of bladder-derived tumors in china is outside 10, for 36cancers in 185coordinates.ca..

For first-line treatment of unresectable locally advanced or metastatic urothelial cancer, such as cisplatin resistance, cisplatin-containing chemotherapy regimens such as GC (gemcitabine + cisplatin), MVAC (methotrexate + vinblastine + doxorubicin + cisplatin) are the first-line regimens. Patients who are intolerant to cisplatin may consider carboplatin-containing regimens, such as the gemcitabine + carboplatin regimen. Other patients may use PD-1 inhibitors such as Pabolizumab or altertib, and chemotherapeutic agents such as gemcitabine + paclitaxel or gemcitabine unit, etc. PD-1 inhibitors are preferentially recommended among patients with PD-L1 expression. For second-line treatment of patients, the current therapeutic approaches include immunotherapy, targeted therapy, ADC drug therapy and chemotherapy. At present, three or more lines of medicines have no standard treatment, and corresponding chemotherapy, targeted therapy and biological therapy need to be selected according to the using condition of the medicines at the front line.

In conclusion, patients who have failed platinum-containing chemotherapy regimens and PD-1 inhibitors are subsequently lack of effective therapeutic approaches and need to explore new drugs.

The incidence rate and the mortality rate of breast cancer in female malignant tumors of China are the first place, and the health of the female in China is seriously threatened (Zheng Rong longevity, sunkuoxin, zhang Xin Wei, etc.. 2015, the prevalence situation of the Chinese malignant tumors is analyzed [ J ]. The journal of China tumor, 2019, 41 (1): 19-28 ]. In 2015, about 30.4 thousands of breast cancer cases of new women in China occur, the morbidity is 45.29/10 thousands, and 7 thousands of cases of death occur. About 3% to 10% of new cases of breast cancer present distant metastasis at the time of diagnosis, while 30% of early cases develop advanced breast cancer. The 5-year survival rate of advanced breast cancer is only 20% (national cancer control center breast cancer expert committee, chinese advanced breast cancer standard medical guidelines (2020 edition).

Chemotherapy is an indispensable treatment for hormone receptor negative, HER-2 positive, postoperative disease-free survival shorter (< 2 years), faster tumor development, marked symptoms, extensive visceral metastasis, hormone receptor positive, endocrine therapy abortive in non-surgical locally advanced, recurrent or metastatic breast cancer. In the former chemotherapy patients (including adjuvant chemotherapy), the first-line chemotherapy is preferably based on anthracycline and/or taxoid. Metastatic breast cancer in which the anthracyclines failed or approached cumulative doses and have not been previously treated with taxanes, first-line chemotherapy prefers either a taxoid-based regimen or a taxoid-single-dose regimen. Adjuvant treatment of taxus, which can be reused for >1 year from relapse time, is preferred over the choice of other drugs, which are not used during adjuvant treatment and treatment phases. For recurrent metastatic breast cancer that fails both the preoperative and adjuvant therapies for anthracyclines and taxanes, there are currently no standard chemotherapeutic regimens, and the drugs that can be considered are capecitabine, vinorelbine, gemcitabine, platins, eribulin, uliduron, another taxus species (e.g., albumin paclitaxel), and liposomal doxorubicin, and single or combination regimens can be considered, thus there is still a need to continue the development and exploration of new drugs for back-line therapy.

Classic osteosarcoma is the most common primary malignant tumor of bone, and the annual incidence rate is about 2-3/100 ten thousand, and accounts for only 0.2% of human malignant tumor. Osteosarcoma can occur in any age group, preferably in adolescents, with about 75% of patients having an age of between 15 and 25 years, with a median age of 20 years, less than 6 years or more than 60 years being relatively rare. Men are more sick than women, with a ratio of about 1.4:1, this difference was particularly apparent before the age of 20. There is clear lung metastasis in about 20% of initial diagnoses, and in about 30% of patients lung metastasis occurs within 1 year after definitive diagnosis of osteosarcoma. Before the 70's of the 20 th century, surgery was the only treatment for osteosarcoma, with amputation predominating and an average 5-year survival rate of only 19.7%. A new treatment scheme of adjuvant chemotherapy, surgery and adjuvant chemotherapy is formed in the early 70 s, the survival rate in 5 years is remarkably improved to 60-80% (Daxiaohui, the current situation and prospect of the comprehensive treatment of Chinese osteosarcoma [ J ]. China anatomy and clinical journal, 2019 (01): 1-5.), and the traditional Chinese medicine is continuously used. Whereas the survival rate of patients with metastatic or recurrent osteosarcoma has been nearly unchanged over the last 30 years, with an overall 5-year survival rate of about 20% (Meyers P A, healey J H, chou A J, et al.Addition of pamidrate to chemotherapy for the treatment of osteoarcoma [ J ]. Cancer,2011,117 (8): 1736-1744.).

There are numerous, but not specific, chemotherapy regimens for osteosarcoma internationally, but the therapeutic effects are similar due to the similar type and dose strength of the drugs used. Doxorubicin, cisplatin, ifosfamide and large doses of methotrexate are listed as first-line chemotherapy drugs for osteosarcoma, and the same drugs are also used in neoadjuvant chemotherapy. More than two drugs are selected for each patient and sufficient dosage strength is ensured. Compared with the two-drug combination scheme, the three-drug combination scheme has 58% and 48% of EFS (non-time survival) rate in 5 years and 70% and 62% of OS rate in 5 years respectively. After first-line chemotherapy fails in osteosarcoma patients, participation in clinical trials is an opportunity to obtain better efficacy or more recent treatment due to the temporary absence of a second-line treatment regimen that benefits overall survival.

In summary, chemotherapy is an important means of clinical treatment in addition to surgical treatment at present. The prognosis of patients with advanced or unresectable primary chemotherapy failure is poor, the medical evidence of targeted and immunotherapy for osteosarcoma is insufficient, and the seeking or adoption of a new cytotoxic drug or targeted drug therapy brings a chance for secondary therapy.

Soft tissue sarcomas refer to a group of malignancies originating from non-epithelial extraosseous tissue, accounting for approximately 0.8% of all human malignancies, having an annual disease rate of approximately 3.4/10 million in the United states, 4-5/10 million in Europe, and an annual disease rate of approximately 2.38/10 million in China (Siegel R L, miller K D, fuchs H E, et al Cancer statics, 2021[ J ]. CA: A Cancer Journal for Clinicians,2021,71 (1)). Patients of any age can develop the disease, the proportion of the number of the male and female patients in China is close to 1, the incidence rate is obviously increased along with the increase of the age, and the incidence rate at the age of 80 years is about 8 times of that at the age of 30 according to the incidence rate after age calibration (Burningham Z, hashbe M, spector L, et al. The epidemiology of Sarcoma [ J ]. Clin Sarcoma Res,2012,2 (1): 14). Soft tissue sarcomas are derived primarily from mesoderm and partially from neuroectoderm and consist primarily of: muscle, fat, fibrous tissue, blood vessels, peripheral nerves, and the like. The most common sites of occurrence are the extremities (about 53%), followed by retroperitoneum (19%), torso (12%), head and neck (11%). The tissue can be divided into 12 types according to tissue sources, and more than 50 types of subtypes can be divided according to different morphologies and biological behaviors. The common subtypes are: undifferentiated polymorphic sarcoma, liposarcoma, leiomyosarcoma, synovial sarcoma. The most common soft tissue sarcoma in adolescents and children is rhabdomyosarcoma.

Soft tissue sarcoma is mainly characterized by gradual growth of painless masses, strong hiding property, disease course from months to years, and pain, numbness and even edema of limbs when tumor gradually increases and compresses nerves or blood vessels. In some cases, the lump will rapidly increase in a short period, the skin temperature will rise, and the regional lymph nodes will swell. Caution is needed to be given to the possibility of elevated tumor levels. Soft tissue sarcoma has high malignancy degree, and has the characteristics of short course of disease, early occurrence of blood circulation metastasis, easy recurrence after treatment, etc. The most common metastatic site for sarcoma of the extremities is the lungs, while retroperitoneal and gastrointestinal sarcomas are most commonly metastasized to the liver. The total five-year survival rate of soft tissue sarcoma is about 60-80%.

Soft tissue sarcomas employ a combination of surgical treatment strategies. Radiotherapy, chemotherapy or concurrent radiotherapy and chemotherapy are considered to be one of the most effective treatment modalities. With the development of a tumor combined treatment mode, preoperative treatment, surgery and chemotherapy are gradually developed, and the method has the advantages of reducing the tumor volume, increasing the limb protection chance, improving the local control rate, reducing the risk of distant metastasis and relapse and improving the survival rate of patients. Chemotherapy sensitivity is an important criterion for selecting chemotherapy for soft tissue sarcomas. Anthracycline-based regimens are preferred in neoadjuvant and adjuvant therapy. While in patients with soft tissue sarcoma of high pathological grade, local recurrence occurs in 40-50% of patients after surgery, and distant metastasis occurs in >50% of patients. Chemotherapy adopted by patients who have metastasis or relapse and cannot completely excise the tumor is palliative chemotherapy, and the purpose of the chemotherapy is to reduce and stabilize the tumor, relieve symptoms, prolong life cycle and improve life quality.

The formulation of palliative chemotherapy for metastatic or recurrent unresectable tumors will vary from person to person. Patients with metastatic, non-polymorphic rhabdomyosarcoma, the chemotherapy regimen should be selected to be alternating VAC/VI/VCD/IE in high-risk groups. Whereas for non-finger soft tissue sarcoma, doxorubicin and ifosfamide are the cornerstone drugs. There is currently no accepted second-line chemotherapy regimen for non-finger soft tissue sarcomas.

In conclusion, the malignancy degree of the soft tissue sarcoma is high, and for a soft tissue sarcoma patient who fails the first-line treatment, the types of selectable second-line treatment medicines are limited, the curative effect is limited, and the participation in clinical trials is one of recommended schemes. Improving the treatment of patients with soft tissue sarcoma and exploring new drugs become urgent needs in clinic.

Mitoxantrone hydrochloride is an anthraquinone chemotherapeutic drug, and the indications of the common injection liquid approved by FDA for mitoxantrone hydrochloride are multiple sclerosis, prostatic cancer and acute myelogenous leukemia; is approved in China to be suitable for patients with lymphoma, leukemia, breast cancer and the like, and the recommended dose is 12-14mg/m for single-medicine adult ² Once every 3-4 weeks; or 4-8mg/m ² Once a day, continuously using for 3-5 days at an interval of 2-3 weeks; the combined dosage is 5-10mg/m ² Once. Mitoxantrone as an anthracycline drug has major adverse reactions mainly manifested as cardiotoxicity, bone marrow suppression, and gastrointestinal reactions. The bone marrow suppression and the gastrointestinal reaction can be solved by administering proper medicines, but the cardiotoxicity often brings serious consequences, and is the most serious adverse reaction of the anthracycline, and clinical research and practical observation show that most of the cardiotoxicity caused by the anthracycline is progressive and irreversible, and particularly the heart injury is easily caused by the first use of the anthracycline. Chronic dose-accumulating limiting toxicity-cardiotoxicity is a common concern for clinicians. Thus, mitoxantrone has not been recommended as a treatment for breast cancer by the authoritative clinical guidelines, although this indication is approved. However, mitoxantrone has never been approved for urothelial carcinoma, bone and soft tissue sarcomas.

Drug sensitivity varies from tumor to tumor. Previous studies have shown that the same drug may be administered in different regimens for different indications. For example Doxil (Doxil) is approved by FDAThree indications, respectively: (1) Ovarian cancer, recommended dose is 50mg/m ² Once every 4 weeks intravenous administration; (2) Kaposi's sarcoma at a recommended dose of 20mg/m ² Once every 3 weeks intravenous administration; (3) Multiple myeloma at a recommended dose of 30mg/m ² Bortezomib was administered intravenously the fourth day after administration. Also as Abraxane (injectable paclitaxel [ albumin bound form)]) Approved in the FDA are also three indications, (1) metastatic breast cancer: recommended dose is 260mg/m ² Intravenous drip for 30 minutes, administered once every 3 weeks; (2) non-small cell lung cancer: recommended dose is 100mg/m ² Intravenous drip for 30 minutes, one treatment course every 21 days, and administration is carried out on days 1, 8 and 15 respectively; carboplatin was administered immediately after paclitaxel (albumin-bound) injection on day 1, once every 21 days; (3) pancreatic cancer: the recommended dosage is 125mg/m ² The gemcitabine was administered once on each of days 1, 8 and 15 in a 28-day cycle by intravenous drip for 30 to 40 minutes, and immediately after each administration of paclitaxel for injection (albumin-bound type). The starting doses for AmBisome (amphotericin B liposomes for injection) for the following indications were again: (1) empirical treatment: the recommended dose is 3 mg/kg/day; (2) Systemic fungal infections (aspergillus, candida, cryptococcus): the recommended dosage is 3-5 mg/kg/day; (3) cryptococcal meningitis in HIV-infected persons: recommended dosage is 6 mg/kg/day (days 1-5), 3 mg/kg/day (days 4, 21); (4) visceral leishmaniasis patients with immunocompromised: 4 mg/kg/day (days 1-5), 4 mg/kg/day (days 10, 17, 24, 31, 38); dosage and administration data are customized according to the actual condition of a patient so as to achieve maximum drug effect and minimum toxicity or adverse reaction. It can be seen that the safe and effective dosages for the same drug for different indications vary. The dosage and administration data should be customized according to the specific disease species and the actual condition of the patient, so as to achieve the maximum drug effect and the minimum toxicity or adverse reaction, and obtain the effect of safely and effectively treating the disease. The mitoxantrone liposome is a special dosage form different from a common injection, the absorption, distribution and metabolism conditions after entering the body are very complicated, and the simple derivation from one indication to another is difficult for the treatment of different indications, particularly the treatment of different tumorsThe indications are different.

Therefore, it is necessary to perform a systematic study on whether the mitoxantrone liposome is suitable for the treatment of urothelial cancer, breast cancer, and bone and soft tissue sarcoma, so as to define the safe and effective dose of the mitoxantrone liposome and provide reference for clinical treatment.

Disclosure of Invention

The invention provides application of mitoxantrone hydrochloride liposome in preparing a medicament for treating urothelial carcinoma, breast cancer, bone and soft tissue sarcoma.

Preferably, the urothelial cancer is locally advanced or metastatic urothelial cancer, more preferably, the urothelial cancer is locally advanced or metastatic urothelial cancer that has failed a platinum-containing chemotherapy regimen and/or PD-1 inhibitor treatment; alternatively, locally advanced or metastatic urothelial cancer that failed treatment with a platinum-containing chemotherapeutic regimen, but was rejected for treatment with a PD-1 inhibitor; alternatively, locally advanced or metastatic urothelial cancer 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, a HER-2 negative breast cancer that is hormone receptor negative, or, a HER-2 negative breast cancer that is hormone receptor positive but not suitable for endocrine therapy or endocrine therapy resistance; alternatively, HER-2 negative breast cancer that has failed anthracycline and/or taxane-based drug therapy; preferably, the HER-2 negative breast cancer comprises immunohistochemical HER-2 0 or 1+ and immunohistochemical HER-2 + is a breast cancer that is confirmed to be negative by in situ hybridization.

Preferably, the bone and soft tissue sarcoma is advanced bone and soft tissue sarcoma, further preferably metastatic or locally advanced bone and soft tissue sarcoma that failed at least first line therapy.

Preferably, mitoxantrone hydrochloride liposomes are used as the sole active ingredient for the preparation of a medicament for the treatment of urothelial cancer, breast cancer, bone and soft tissue sarcomas.

Preferably, the medicament is in injection dosage forms, including liquid injection, powder for injection, tablets for injection and the like.

Preferably, the medicament is a liquid injection.

According to an embodiment of the invention, when the drug is a liquid injection, the drug contains 0.5-5mg/ml, preferably 1-2mg/ml, more preferably 1mg/ml, mitoxantrone based on mitoxantrone.

The invention also provides a method of treating urothelial cancer, breast cancer, bone and soft tissue sarcomas by administering to a patient in need thereof a therapeutically effective amount of mitoxantrone hydrochloride liposomes.

The invention also provides the application of the mitoxantrone hydrochloride liposome in treating urothelial carcinoma, breast cancer, bone and soft tissue sarcoma.

Preferably, the urothelial cancer is locally advanced or metastatic urothelial cancer; more preferably, the urothelial cancer is a locally advanced or metastatic urothelial cancer that has failed a platinum-containing chemotherapy regimen and/or a PD-1 inhibitor, or a locally advanced or metastatic urothelial cancer that has failed a platinum-containing chemotherapy regimen but has been denied treatment with a PD-1 inhibitor, or a locally advanced or metastatic urothelial cancer 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, a HER-2 negative breast cancer that is hormone receptor negative, or, a HER-2 negative breast cancer that is hormone receptor positive but not suitable for endocrine therapy or endocrine therapy resistance; alternatively, HER-2 negative breast cancer that has failed anthracycline and/or taxane-based drug therapy; preferably, the HER-2 negative breast cancer comprises immunohistochemical HER-2 0 or 1+ and immunohistochemical HER-2 + is negative breast cancer determined by in situ hybridization;

preferably, the bone and soft tissue sarcoma is an advanced bone and soft tissue sarcoma, further preferably a metastatic or locally advanced bone and soft tissue sarcoma that has failed at least one line of treatment.

Preferably, the therapeutically effective amount is a mitoxantrone dose of 8-30mg/m ² More preferably 12 to 20mg/m ² Or 16-30mg/m ² . Specifically, for example, 12mg/m ² ，14mg/m ² ，16mg/m ² ，18mg/m ² ，20mg/m ² 。

Preferably, the mode of administration according to the invention is intravenous. Preferably, the dosing cycle is once every 3 weeks. Preferably, the treatment administered to the patient is 6-8 cycles. Preferably, the administration time of the liposome pharmaceutical preparation by instillation is 30min to 120min, preferably 60min to 120min, and further preferably 60 ± 15min per intravenous administration.

The invention also provides mitoxantrone hydrochloride liposome which is used for treating urothelial carcinoma, breast cancer, and sarcoma of bone and soft tissue of a patient.

In some embodiments, the liposomes are in injectable dosage forms, including liquid injections, injectable powders, injectable tablets, and the like. When the medicine is liquid injection, the liposome contains mitoxantrone 0.5-5mg/ml, preferably 1-2mg/ml, and more preferably 1mg/ml.

In some embodiments, the liposomes are used alone to treat urothelial cancer, breast cancer, bone and soft tissue sarcomas in a patient.

In some embodiments, the therapeutically effective amount of the liposomes (in terms of mitoxantrone) is 8-30mg/m ² More preferably 12 to 20mg/m ² Or 16-30mg/m ² . Specifically, for example, 12mg/m ² ，14mg/m ² ，16mg/m ² ，18mg/m ² ，20mg/m ² 。

In some embodiments, the liposome is administered intravenously. Preferably, the dosing cycle is once every 3 weeks. Preferably, the treatment administered to the patient is 6-8 cycles. Preferably, the administration time of the liposome by instillation is 30min to 120min, preferably 60min to 120min, and further preferably 60 +/-15 min for each intravenous administration.

In the context of the present invention, the dosages are in terms of mitoxantrone unless otherwise specified.

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

In some embodiments, the drug or mitoxantrone hydrochloride liposome has one or more of the following properties:

(i) The mitoxantrone hydrochloride liposomes have a particle size of about 30-80nm, e.g., about 35-75nm, about 40-70nm, about 40-60nm, or about 60nm;

(ii) Mitoxantrone hydrochloride forms a poorly soluble precipitate with multivalent counter ions (e.g., sulfate, citrate, or phosphate) within the liposomes;

(iii) The phospholipid bilayer of the mitoxantrone hydrochloride liposome contains phospholipids having a phase transition temperature (Tm) above body temperature, such that the liposome has a phase transition temperature above body temperature, e.g., the phospholipids are selected from the group consisting of hydrogenated soy lecithin, phosphatidylcholine, hydrogenated egg yolk lecithin, dipalmitoyl lecithin, distearoyl lecithin, or any combination thereof;

(iv) The phospholipid bilayer of the mitoxantrone hydrochloride liposome contains hydrogenated soybean lecithin, cholesterol and polyethylene glycol 2000 modified distearoyl phosphatidyl ethanolamine (DSPE-PEG 2000).

(iii) the phospholipid bilayer of the mitoxantrone hydrochloride liposome contains hydrogenated soybean lecithin, cholesterol, and polyethylene glycol 2000-modified distearoyl phosphatidyl ethanolamine at a mass ratio of about 3.

(iiiv) mitoxantrone hydrochloride liposome is the mitoxantrone hydrochloride liposome of the national standard H20220001.

In the context of the present invention, the mitoxantrone hydrochloride liposomes, having a particle size of about 30 to 80nm, comprise: 1) Mitoxantrone, an active ingredient, which forms a poorly soluble precipitate with multivalent counter ions within liposomes, 2) phospholipid bilayers contain phospholipids with a phase transition temperature (Tm) above body temperature, whereby the phase transition temperature of the liposomes is above body temperature. The phospholipid with Tm higher than body temperature is phosphatidyl choline, hydrogenated soybean lecithin, hydrogenated yolk lecithin, dipalmitin lecithin or distearyl lecithin or any combination thereof, and the particle size is about 35-75nm, preferably 40-70nm, more preferably 40-60nm, and particularly preferably 60nm. Preferably, the phospholipid bilayer contains hydrogenated soybean lecithin, cholesterol and polyethylene glycol 2000 modified distearoyl phosphatidyl ethanolamine, the mass ratio is 3. Preferably, the phospholipid bilayer of the liposome comprises hydrogenated soybean lecithin, cholesterol and polyethylene glycol 2000 modified distearoyl phosphatidyl ethanolamine, the mass ratio is 3: chol: DSPE-PEG2000: the weight ratio of mitoxantrone is 9.58.

In the context of the present invention, mitoxantrone liposomes are prepared as follows: HSPC (hydrogenated soybean lecithin), chol (cholesterol), and DSPE-PEG2000 (polyethylene glycol 2000-modified distearoylphosphatidylethanolamine) were weighed in a mass ratio of (3. Mixing the ethanol solution of phospholipid with 300mM ammonium sulfate solution, and shaking and hydrating at 60-65 ℃ for 1h to obtain the heterogeneous multi-chamber liposome. The particle size of the liposomes is then reduced using a microfluidizer. The obtained sample was diluted 200 times with 0.9% NaCl solution and the average particle size of the particles was about 60nm, and the main peak was concentrated between 40-60nm, as detected by NanoZS. The blank liposome external phase was then removed of ammonium sulfate using an ultrafiltration device and the external phase was replaced with 290mM sucrose and 10mM glycine to form a transmembrane ammonium sulfate gradient. According to the ratio of lipid to drug of 16. After about 1h of incubation, the encapsulation efficiency was about 100% as demonstrated using gel exclusion chromatography. Wherein HSPC: chol: DSPE-PEG2000: mitoxantrone weight ratio 9.58.

It should be understood that numerous technical details and parameters of the above-described exemplary methods of preparation can be set forth and determined within the scope of one of ordinary skill in the art. For example, the amino acid species that are replaced by glycine in the outer phase used to form the transmembrane ammonium sulfate gradient include, but are not limited to, histidine, asparagine, glutamic acid, leucine, proline, alanine. For another example, the mass ratio of HSPC, chol and DSPE-PEG2000 can be adjusted appropriately. Also for example, for preparing lipid-drug ratio parameters in a particular liposomal pharmaceutical formulation, one skilled in the art can design, test, and ultimately arrive at a suitable lipid-drug ratio to maximize drug loading while reducing drug leakage, for mitoxantrone hydrochloride liposomal formulations of the present application, a wide range of lipid-drug ratios can be used, e.g., as low as 2:1 is possible, and more suitably the 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. thus, the several advantageous properties of the mitoxantrone hydrochloride liposomal formulations described above are more important and the methodologies for achieving these properties are varied.

The first-line, second-line or more than second-line drug for treating urothelial cancer, breast cancer, and bone and soft tissue sarcoma of the present invention refers to the first-line, second-line or more than second-line drug approved by the drug administration in china or abroad (e.g., usa, eu, japan, korea, etc.) for treating urothelial cancer, breast cancer, and bone and soft tissue sarcoma, and includes but is not limited to: FDA approved taxoids, PD-1 inhibitors, ifosfamide, etc.

Advantageous effects

The research of the applicant of the invention unexpectedly discovers that the mitoxantrone hydrochloride liposome has better treatment effect on urothelial carcinoma, breast cancer, bone and soft tissue sarcoma and can be used for preparing related medicaments.

Detailed Description

The technical solution of the present invention will be further described in detail with reference to specific embodiments. It is to be understood that the following examples are only illustrative and explanatory of the present invention and should not be construed as limiting the scope of the present invention. All the technologies realized based on the above-mentioned contents of the present invention are covered in the protection scope of the present invention.

Unless otherwise indicated, the raw materials and reagents used in the following examples are all commercially available products or can be prepared by known methods.

The following abbreviations are used in the present invention:

CR: complete remission is defined specifically according to the "solid tumor efficacy evaluation criteria (RECIST 1.1)" commonly used in international research.

PR: partial remission is defined specifically according to the "solid tumor efficacy evaluation criteria (RECIST 1.1)" commonly used in international research.

SD: the disease stability is specifically defined according to the general "evaluation criteria for solid tumor efficacy (RECIST 1.1)" in international research.

PD: the disease progression is specifically defined according to the "evaluation criteria for solid tumor efficacy (RECIST 1.1)" commonly used in international research.

Objective Remission Rate (ORR) = (CR + PR)/total number of evaluable cases 100%.

Disease Control Rate (DCR) = (CR + PR + SD)/total number of evaluable cases 100%.

In order to examine the effect of mitoxantrone hydrochloride liposome on treating urothelial carcinoma, breast cancer, and bone and soft tissue sarcoma, the inventors developed clinical studies. The mitoxantrone hydrochloride liposome can effectively treat urothelial cancer, breast cancer, bone and soft tissue sarcoma, and has better curative effect and less adverse reaction compared with the common mitoxantrone hydrochloride injection. The mitoxantrone hydrochloride liposome injection used in the following examples was provided by Shiyao Kagaku (Shijiazhuang) Co., ltd. (national Standard H20220001).

Example 1 phase II clinical trial to evaluate the effectiveness and safety of mitoxantrone hydrochloride liposomal injection for the treatment of unresectable locally advanced or metastatic urothelial cancer

This is an open, multicenter phase II study, where the included subjects will receive mitoxantrone hydrochloride liposomal injection therapy to evaluate the effectiveness and safety of mitoxantrone hydrochloride liposomal injection in unresectable locally advanced or metastatic urothelial cancer.

1. Design of experiments

1. Test procedure

Group 40 patients with no excisionIn a subject with locally advanced or metastatic urothelial cancer. The study population was predominantly subjects who failed the platinum-containing chemotherapy regimen and the PD-1 inhibitor treatment (including not receiving PD-1 inhibitor treatment); is intolerant to cisplatin, and needs to be treated systemically. The study included a screening phase, a treatment phase and a follow-up phase. The screening period was 28 days, and eligible subjects were screened for treatment. The mitoxantrone hydrochloride liposome injection is administered at 20mg/m in the treatment period ² Single-dose treatment, one cycle every 3 weeks (q 3 w), 6 cycles on the first day of the cycle. After the treatment is finished, the follow-up period is started. The follow-up period required safety follow-up (28 days after the last administration) and survival follow-up (every 6 weeks after the last administration). Safety evaluations were performed from the first dose to 28 days after the last dose, and were performed before each cycle to determine whether to dose the next cycle. Tumor evaluation was performed every two cycles throughout the study period according to RECIST1.1 criteria until disease progression, death or receipt of new anti-tumor therapy.

2. Duration of study

The study included a screening period of 4 weeks (28 days), a treatment period of 6 cycles (18 weeks), a last safety follow-up, 4 weeks (28 days) after dosing was completed, followed by a follow-up visit for survival every 6 weeks, with each patient lasting approximately 12 months.

40 subjects were enrolled in the study and the whole period of the study ranged from 18 to 24 months.

2. The test population is as follows:

subjects who met all of the following inclusion criteria and did not have any exclusion criteria were eligible for inclusion in the candidate clinical study.

(one) inclusion criteria

1) The subject voluntarily participated in the study and signed an informed consent;

2) The age is more than or equal to 18 years old, and the male and the female are not limited;

3) Histologically confirmed urothelial cancer;

4) Subjects who have failed a platinum-containing chemotherapy regimen and treatment with a PD-1 inhibitor (including not receiving PD-1 inhibitor treatment); the medicine is intolerant to cis-platinum and needs to be treated systemically;

5) At least one evaluable lesion at baseline that meets RECIST1.1 definition;

6) ECOG score is 0-1;

7) Subject laboratory test values met the following requirements:

the absolute value (ANC) of neutrophil at least 1.5x10 ⁹ L (no G-CSF leukocyte treatment received within 1 week prior to laboratory examination);

hemoglobin (Hb) > 9.0g/dL (no infusion of erythrocytes within 1 week before laboratory examination);

platelet ≥ 75x10 ⁹ L (no platelets were infused within 1 week prior to laboratory examination);

creatinine no greater than or equal to 1.5x ULN;

total bilirubin < 1.5x ULN (for liver metastases < 3x ULN);

alanine Aminotransferase (AST)/aspartate Aminotransferase (ALT) of less than or equal to 3x ULN (for liver transfer subjects, less than or equal to 5x ULN);

coagulation function: prothrombin Time (PT), international Normalized Ratio (INR) ≦ 1.5x ULN (INR ≦ 3 for receiving anticoagulant therapy such as warfarin);

8) Subjects and their partners take effective contraceptive measures during the trial period to within 6 months after the end of the last dose (e.g.: combined hormones (containing estrogen and progestogen) to inhibit ovulation, progestogen to inhibit ovulation, intrauterine devices, intrauterine hormone release systems, bilateral vasectomy, bilateral tubal ligation, sexual behavior avoidance, etc.); female subjects are urine or blood HCG negative (except for menopause and hysterectomy);

(II) exclusion criteria

1) Severe allergy to mitoxantrone or liposomal drugs;

2) A brain or meningeal transfer subject;

3) A prior allogeneic organ transplant or allogeneic bone marrow transplant;

4) Survival <12 weeks;

5) Patients with active hepatitis B (HBsAg or HBcAb positive and HBV DNA more than or equal to 2000 IU/mL), active hepatitis C (HCV antibody positive and HCV RNA higher than the lower limit of the detection value of the research center) and HIV antibody positive;

6) Active bacterial or fungal or viral infections requiring intravenous infusion within 1 week prior to first drug administration;

7) Those who have received any anti-tumor therapy within 4 weeks before the first administration (2 weeks before the administration of the Chinese medicinal material or the Chinese patent medicine);

8) Other clinical trial medications were taken within 4 weeks prior to the first administration;

9) Major surgery was received within 3 months prior to the first dose, or major operators were planned during the study;

10 Presence of previous antitumor therapy toxicity > level 1 (except for alopecia, pigmentation, or other toxicity in the study considered to be a non-safety risk to the subject);

11 Severe thrombotic or embolic events such as cerebrovascular accidents (including transient ischemic attacks), pulmonary embolism, etc. occurred within the past 6 months;

12 Other malignant active tumors had been developed within 3 years ago; in addition to locally treatable cancers that have cured, such as basal or squamous cell skin cancer or prostate, cervical or breast cancer in situ;

13 Cardiac dysfunction, including:

long QTc syndrome or QTc interval >480ms;

complete left bundle branch block, II-or III-degree atrioventricular block (except after pacemaker implantation therapy);

severe, uncontrolled arrhythmias requiring drug treatment;

history of chronic congestive heart failure, NYHA ≥ grade 3;

heart ejection fraction less than 50% within 6 months;

CTCAE > grade 2 valvular heart disease;

uncontrolled hypertension (defined as multiple measurements of systolic >150mmHg or diastolic >100mmHg with drug control);

myocardial infarction, unstable angina, history of severe pericardial disease, and electrocardiographic evidence of acute ischemic or severe conduction abnormalities within 6 months prior to screening.

14 Has received doxorubicin or other anthracyclines and has a cumulative doxorubicin dose of over 350mg/m ² (anthracycline equivalent dose calculation: 1mg doxorubicin =2mg epirubicin =2mg pirarubicin =0.5mg demethoxydaunorubicin =0.45mg mitoxantrone; except doxorubicin liposomes);

15 Pregnant or lactating women;

16 Have any serious and/or uncontrollable disease that may affect other diseases that the patient may have in the study (including, but not limited to, diabetes mellitus, renal disease requiring dialysis, severe liver disease, life threatening autoimmune and hemorrhagic disease, drug abuse, neurological disease, etc.) as judged by the investigator;

17 Other investigators judged not to be suitably attending.

(III) withdrawal from treatment and withdrawal from study standards

The subjects had to stop receiving the test medication during the study if either:

1) The subject has intolerable toxicity, and the researcher considers that the risk of continuously receiving the test drug treatment is greater than the benefit;

2) Concomitant disease that occurs during treatment does not allow subsequent follow-up;

3) Disease progression assessed by imaging;

4) Clinical assessment of disease progression or occurrence of major protocol violations, or poor subject compliance, and the investigator determines that continued treatment with the test drug would not benefit;

5) A subject is pregnant;

6) Death;

7) Any of the withdrawal criteria were met.

All subjects who dropped out of treatment continued follow-up on the study schedule, except subjects who stopped treatment due to death or who met any of the criteria for dropping out of the study.

The subject is entitled to withdraw from the study at any time for any reason. Subjects will exit the study procedure when either of the following occurs:

1) Loss of visit;

2) The subject or family (when the subject himself cannot make a decision) requires withdrawal from the trial;

3) Termination of the study;

4) And others.

3. Results of the study

Subjects who have failed the platinum-containing chemotherapy regimen and the PD-1 inhibitor treatment (including those who do not receive the PD-1 inhibitor treatment) and subjects who are not resistant to cisplatin and who have undergone systemic treatment subsequently lack effective treatment means, and a new drug is urgently needed to be searched. The inventor of the invention uses the mitoxantrone liposome, the drug has the functions of slow release, targeting, toxicity reduction and synergy after entering the human body through intravenous infusion, and compared with the common mitoxantrone injection, the liposome preparation can adopt higher dosage, thereby not only improving the effectiveness, but also reducing the occurrence probability of adverse reaction, and obtaining ideal treatment effect only by using single-drug therapy.

The study of the present invention showed that, of at least 4 subjects (2 subjects treated for 2 cycles, 2 subjects treated for 1 cycle), SD 2 subjects (1 subject treated for 1 cycle, 1 subject treated for 2 cycles), DCR 50% (2/4). Mitoxantrone liposomes have good therapeutic effects on urothelial cancer patients, particularly patients who have failed platinum-containing chemotherapy regimens and PD-1 inhibitor therapy. The drug was continuously administered to 2 subjects evaluated as SD, and a better therapeutic effect was obtained.

The scheme improves the treatment scheme of the urothelial cancer, lays a foundation for combined medication to impact first-line and second-line treatment, changes the traditional treatment mode of the urothelial cancer, and expects the application of similar high-efficiency and low-toxicity chemotherapeutic drugs in the field of advanced solid tumors.

Example 2 phase II clinical trial of mitoxantrone hydrochloride liposome injection for the treatment of HER-2 negative advanced breast cancer

This is an open, multicenter phase II study, where included subjects will receive mitoxantrone hydrochloride liposome injection therapy to evaluate the safety and efficacy of mitoxantrone hydrochloride liposome injection therapy for HER-2 negative advanced breast cancer.

1. Design of experiments

1. Test procedure

The study is divided into a screening period, a treatment period and a follow-up period. The screening period was 28 days, and eligible subjects were screened for treatment. During the treatment period, the subject receives mitoxantrone hydrochloride liposome injection of 20mg/m ² 3 weeks is a cycle, 6 cycles, and after completion of 6 cycles of treatment, investigators and sponsors will decide whether to continue dosing based on subject benefit and risk. Follow-up included treatment termination visit (28 days after last dose) and survival follow-up (every 6 weeks after treatment termination visit), with all subjects on the phone or visit for survival follow-up until subject death.

Tumor evaluation: tumor assessments were performed every 2 cycles during the treatment period and every 6 weeks during the follow-up period, until disease progression or initiation of a new anti-tumor therapy, according to RECIST1.1 criteria.

And (3) safety evaluation: completing vital signs, physical examination, weight, ECOG physical strength score, 12-lead electrocardiogram, echocardiography and laboratory examination before and after treatment in each period. Routine examinations were performed for D8 and D15 blood per cycle.

2. Duration of study

The study included a screening period of 4 weeks (28 days), a treatment period of 6 cycles (18 weeks), an end of treatment visit (28 days after the last dose), followed by a follow-up visit for survival every 6 weeks until patient failure or death, each patient lasting about 12 months.

The study included 73 subjects, and the duration of the study was approximately 36 months.

2. The test population:

subjects who met all of the following inclusion criteria and did not have any exclusion criteria were eligible for inclusion in the candidate clinical study.

(one) inclusion criterion:

the subject must meet all of the following criteria:

1) The subject volunteered to participate in the study and signed an informed consent;

2) Age 18-75 years (including 18 years and 75 years);

3) Histopathologically confirmed HER-2 negative breast cancer (including immunohistochemical HER-2 0 or 1+, immunohistochemical HER-2 + is confirmed to be negative by in situ hybridization);

4) Non-operable locally advanced or recurrent/metastatic breast cancer that progresses after first-line and above chemotherapy;

5) Breast cancer that is hormone receptor negative or hormone receptor positive but not suitable for endocrine therapy or endocrine therapy resistance;

6) The treatment with at least one anthracene nucleus and one taxus medicament (the treatment can be carried out on the subjects who do not use anthracene nucleus medicaments due to high risk factors of cardiac toxicity) in the past, and the treatment with at most four chemotherapy schemes is carried out;

7) At least one measurable lesion at baseline (RECIST 1.1 criteria);

8) ECOG score is 0-2 points;

9) Good organ function (no transfusion or growth factor support treatment received within 2 weeks prior to the first application of study medication) includes:

the absolute value of neutrophil (ANC) is not less than 1.5X10 ⁹ /L；

Hemoglobin (Hb) 90g/L or more;

platelet ≥ 90X 10 ⁹ /L；

Creatinine no more than 1.5 times the upper limit of the normal value;

total bilirubin is less than or equal to 1.5 times the upper limit of the normal value;

alanine Aminotransferase (AST) and aspartate Aminotransferase (ALT) at 3 times the upper limit of normal (liver metastasis patients at 5 times the upper limit of normal);

10 Negative pregnancy test results, subjects in the child-bearing age promises to take effective contraceptive measures or abstinence within 6 months from study initiation to study last dose;

11 Good compliance and a willingness to follow-up.

(II) exclusion standard:

subjects who meet any of the following criteria will be excluded from this trial:

1) Severe allergy to mitoxantrone or liposomes;

2) Other malignant tumors are suffered in 3 years, and the cervical carcinoma in situ, skin basal cell carcinoma or squamous carcinoma which is already radically cured is not included;

3) Brain metastases and membrane subjects;

4) Active hepatitis B (HBsAg positive and HBV DNA is more than or equal to 2000 IU/mL), active hepatitis C (HCV antibody positive and HCV RNA is higher than the lower limit of the detection value of the research center), and HIV antibody positive subjects;

5) Expected survival time < 3 months;

6) Conversion of previously accepted cumulative doses of anthracyclines to doxorubicin>350mg/m ² (anthracycline equivalent dose calculation: 1mg doxorubicin =2mg epirubicin =2mg pirarubicin =2mg daunorubicin =0.5mg demethoxydaunorubicin =0.45mg mitoxantrone, with the exception of doxorubicin liposomes);

7) Both previous anti-tumor treatments did not recover toxicity to grade 1 (except for hair loss, pigmentation, or other toxicity in the study that was considered to be not a safety risk for the subject);

8) Cardiac dysfunction includes:

long QTc syndrome or QTc interval >480ms;

complete left bundle branch block, degree II/degree III atrioventricular block;

severe, uncontrolled arrhythmias requiring drug treatment;

history of chronic congestive heart failure and NYHA > 3;

cardiac ejection fraction below 50% within 6 months prior to screening;

heart valvular disease with CTCAE greater than or equal to grade 3;

myocardial infarction, unstable angina, severe ventricular arrhythmia, a history of severe pericardial disease, electrocardiographic evidence of acute ischemic or active conduction abnormalities within 6 months prior to screening.

9) Uncontrollable hypertension (under the condition of drug control, systolic pressure is more than or equal to 160mmHg or diastolic pressure is more than or equal to 100mmHg after multiple measurements);

10 Active bacterial, fungal, viral infections requiring intravenous infusion therapy within 1 week prior to the first administration;

11 Any anti-tumor treatment (including chemotherapy, radiotherapy, molecular targeted therapy, immunotherapy, endocrine therapy) is received within 4 weeks before the first administration, an immunomodulator is received as adjuvant therapy of malignant tumor within 2 weeks before the first administration, and any anti-tumor Chinese patent medicine (except the Chinese patent medicines for strengthening the body resistance and the Chinese patent medicines for relieving symptoms) is received within 2 weeks before the first administration.

12 Patients who received other clinical study medication within 4 weeks prior to the first dose;

13 Patients who had undergone major surgery within 12 weeks prior to the first dose, or who were scheduled to undergo major surgery during the study;

14 Deep vein thrombosis or arterial embolism in the past 6 months, including but not limited to superior/inferior vena cava thrombosis, lower limb deep vein thrombosis, pulmonary embolism;

15 For lactating women;

16 Have any serious and/or uncontrollable disease that may affect other diseases that the patient may have in the study (including but not limited to diabetes mellitus, renal disease requiring dialysis, severe liver disease, life threatening autoimmune and hemorrhagic disease, drug abuse, neurological disease, etc.) as judged by the investigator;

17 Other investigators judged not to be suitably attending.

(III) withdrawal from treatment and withdrawal from study standards

Subjects had to discontinue study medication during the study in either of the following cases:

1) Subjects developed intolerable toxicity, and researchers considered that the risk of continuing to receive study drug treatment was greater than the benefit;

2) Disease progression assessed by imaging;

3) Clinically assessed disease progression or major protocol violation, or poor subject compliance, which the investigator decides would not benefit from continued study medication;

4) A subject is pregnant;

5) Death;

6) Meet any one of the withdrawal criteria.

All subjects who dropped out of treatment continued follow-up on the study schedule, except subjects who stopped treatment due to death or who met any of the criteria for dropping out of the study.

The subject is entitled to withdraw from the study at any time for any reason. Subjects will exit the study procedure in the presence of either:

1) Loss of visit;

2) Subject withdrawal informed consent or subject or family member request withdrawal from the trial;

3) Termination of the study;

4) And others.

3. Results of the study

New treatments are urgently sought for advanced breast cancer that has previously failed or recurred to anthracyclines and taxanes.

The present study showed that, of at least 20 evaluable cases, PR 6 (1 with 7 cycles, 3 with 6 cycles, 1 with 5 cycles, 1 with 4 cycles), SD 8 (2 with 6 cycles, 2 with 5 cycles, 1 with 4 cycles, 1 with 3 cycles, 2 with 2 cycles). ORR 30% (6/20), DCR 70% (14/20).

The mitoxantrone liposome has good curative effect on patients with breast cancer, especially patients with advanced breast cancer who have failed or recurred treatment by anthracyclines and taxoids. The 14 subjects evaluated for PR and SD were continued to receive a better treatment.

The inventor brings mitoxantrone hydrochloride single drug treatment of recurrent/metastatic breast cancer into previous clinical research, 30 cases respectively exist in a test group and a control group, and the test group is administered with mitoxantrone hydrochloride liposome injection of 20mg/m ² Every 4 weeks is one cycle. The mitoxantrone hydrochloride injection of 14mg/m was administered to the control group ² Every 4 weeks is a cycle. As a result: test groups PR 4, SD 11, ORR13.3% (4/30), DCR 50% (15/30); the control group PR 2, SD 7, ORR 6.7% (2/30), DCR 30% (9/30).

The mitoxantrone liposome is used by the inventor of the invention, and the drug has the functions of slow release, targeting, toxicity reduction and synergy after entering a human body through intravenous infusion, so that the dosage is higher than that of a common injection, and the mitoxantrone liposome is single-drug therapy, thereby improving the effectiveness and reducing the occurrence probability of adverse reactions.

Example 3 mitoxantrone hydrochloride liposome injection for treatment of advanced stage bone and soft tissue sarcoma failed at least first line of treatment phase Ib clinical trial

This is an open, multicenter phase Ib study, in which included subjects will receive mitoxantrone hydrochloride liposome injection therapy to evaluate the safety and efficacy of mitoxantrone hydrochloride liposome injection against metastatic or unresectable bone and soft tissue sarcomas that have failed at least one previous line of therapy.

1. Design of experiments

1. Test procedure

The study included a screening phase, a treatment phase and a follow-up phase.

The screening period was 28 days, and eligible subjects were screened for treatment. The subjects in the treatment period receive the mitoxantrone hydrochloride liposome injection of 20mg/m ² Day 1 treatment, one treatment cycle (q 3 w) for 3 weeks for 6 cycles. For subjects who have completed 6 cycles of treatment, the investigator co-discussed with the sponsor determines whether to continue dosing based on the risk and benefit of the subject. The follow-up period required safety follow-up (28 + -7 days after the last dose) and survival follow-up (every 6 weeks after the last dose).

Safety assessments were performed 28 days after the first dose to the last dose, before each cycle of dosing, to determine whether the next cycle was dosed, tumor assessments were performed according to the rest 1.1 criteria throughout the study period, once every two cycles of treatment, once every 6 weeks of follow-up until disease progression, death or receipt of new anti-tumor treatments.

2. Duration of study

The study included a screening period of 4 weeks (28 days), a treatment period of 6 cycles (18 weeks), a last safety visit, 4 weeks (28 days) after dosing, followed by a survival visit every 6 weeks for approximately 12 months for each patient.

Not less than 50 subjects are enrolled in the study, and the whole period of the study is 24-36 months.

2. The test population is as follows:

subjects who met all of the following inclusion criteria and did not have any exclusion criteria were eligible for inclusion in the clinical study.

(one) inclusion criterion:

the subject must meet all of the following criteria:

1) The subject voluntarily participated in the study and signed an informed consent;

2) The age is more than or equal to 18 years old, and male and female are unlimited;

3) Pathohistologically confirmed, osteosarcomas or non-assigned soft tissue sarcomas (soft tissue sarcoma types excluded: chemotherapy-refractory sarcomas such as embryonal/alveolar rhabdomyosarcoma, ewing's sarcoma, chemotherapy-refractory sarcomas such as alveolar soft tissue sarcoma, extraosseous mucinous chondrosarcoma, sarcomas requiring special treatment such as gastrointestinal stromal tumor, invasive fibromatosis, etc.);

4) Metastatic or unresectable osteosarcoma or soft tissue sarcoma that has previously failed at least one line of treatment;

5) At least one measurable lesion meeting RECIST1.1 definition exists at baseline;

6) ECOG score is 0-1;

7) Laboratory examination values must meet the following criteria:

the absolute value (ANC) of neutrophil not less than 2.0x10 ⁹ L (no G-CSF leukocyte treatment received within 1 week prior to laboratory examination);

hemoglobin (Hb) 110g/L or more (no infusion of red blood cells for 1 week before laboratory examination);

platelet ≥ 100x 10 ⁹ L (no treatment with platelet infusions within 1 week prior to laboratory examination);

creatinine no greater than or equal to 1.5x ULN;

total bilirubin ≤ 1.5x Upper Limit of Normal (ULN) (combined with liver metastasis subjects ≤ 3.0x ULN);

alanine Aminotransferase (AST)/aspartate Aminotransferase (ALT) of less than or equal to 3.0x ULN (pooled liver metastasis subjects, less than or equal to 5.0x ULN);

coagulation function: prothrombin Time (PT), international Normalized Ratio (INR) less than or equal to 1.5x ULN (INR less than or equal to 3 is allowed for the receiving of warfarin and other anticoagulant therapy drugs)

8) Female subjects are urine or blood HCG negative (except for menopause and hysterectomy);

9) The subjects and their partners agree to take effective contraceptive measures during the trial and within 6 months after the end of the last dose (e.g.: combined hormones (containing estrogen and progestogen) for inhibiting ovulation, progestogen for contraception for inhibiting ovulation, intrauterine device, intrauterine hormone release system, bilateral tubal ligation, vasal ligation, sexual behavior avoidance, etc.);

(II) exclusion criteria:

subjects who meet any of the following criteria will be excluded from this trial:

1) Severe allergy to mitoxantrone or liposomal drugs;

2) Brain or meningeal metastases;

3) A prior allogeneic organ transplant or allogeneic bone marrow transplant;

4) Expected survival time <12 weeks;

5) Chronic hepatitis B (HBsAg or HBcAb is positive, HBV DNA is more than or equal to 1000 IU/mL), hepatitis C (HCV antibody is positive, HCV RNA quantitative detection is higher than the lower limit of the detection value of a research center), and HIV antibody is positive;

6) The presence of a previous antitumor therapy toxicity of > grade 1 (except for alopecia, pigmentation, or other toxicity in the study considered to be not a safety risk for the subject);

7) Active bacterial, fungal, viral infections requiring systemic treatment are present within 1 week prior to the first administration;

8) Local or systemic anti-tumor treatment is performed within 4 weeks before the first administration (2 weeks before the administration of the traditional Chinese medicine or the Chinese patent medicine);

9) Other clinical trial medication is used for treating within 4 weeks before the first administration;

10 Severe thrombosis including pulmonary embolism, visceral embolism, deep vein thrombosis, etc. occurred within 6 months;

11 Other malignant active tumors have been present within 3 years ago, except for locally curable cancers that have been cured, such as basal or squamous cell skin cancer, superficial bladder cancer, or prostate, cervical or breast cancer in situ;

12 Cardiac dysfunction, including:

long QTc syndrome or QTc interval >480ms;

complete left bundle branch block, II-or III-degree atrioventricular block (except after pacemaker implantation treatment);

severe, uncontrolled arrhythmias requiring drug treatment;

history of chronic congestive heart failure, and NYHA ≥ grade 3;

left ventricular ejection fraction was below 50% within the previous 6 months;

valvular heart disease with CTCAE not less than grade 3;

uncontrolled hypertension (defined as multiple measurements of systolic pressure ≥ 160mmHg or diastolic pressure ≥ 100mmHg with drug control);

electrocardiographic evidence of myocardial infarction, unstable angina, a history of severe pericardial disease, acute ischaemia or severe active conduction abnormalities within 6 months prior to screening.

13 Has received doxorubicin or other anthracyclines and has a cumulative doxorubicin dose of over 400mg/m ² (anthracycline equivalent dose calculation: 1mg doxorubicin =2mg epirubicin =2mg daunorubicin =0.5mg demethoxydaunorubicin =0.45mg mitoxantrone); the adriamycin liposome accumulated dose exceeds 900mg/m ² ；

14 Pregnant or lactating women;

15 Have any serious and/or uncontrollable disease that may affect other diseases (including, but not limited to, diabetes mellitus, renal disease requiring dialysis, severe liver disease, life threatening autoimmune and hemorrhagic diseases, drug abuse, neurological diseases, etc.) taking part in the study as judged by the investigator;

16 Other investigators judged not to be suitably attending.

(III) withdrawal from treatment and study criteria

The subjects had to stop receiving the test medication during the study if either:

1) The subject has intolerable toxicity, and the researcher considers that the risk of continuing to receive the drug treatment for the test is greater than the benefit;

2) Concomitant disease that occurs during treatment does not allow for subsequent follow-up;

3) Disease progression assessed by imaging;

4) Clinically assessed disease progression or major protocol violation, or poor subject compliance, which is judged by the investigator to be unable to benefit from continued treatment with the test drug;

5) A subject is pregnant;

6) Death;

7) Satisfying any of the withdrawal criteria.

All subjects who dropped out of treatment continued to follow-up on the study schedule, except subjects who stopped treatment due to death or who met any of the following criteria for dropping out of the study.

The subject is entitled to withdraw from the study at any time for any reason. Subjects will exit the study procedure when either of the following occurs:

1) Loss of visit;

2) The subject or family member requires to withdraw from the test;

3) Termination of the study;

4) And others.

3. Results of the study

For the subjects without the chance of radical operation or metastatic bone and soft tissue sarcoma, after the first-line standard treatment, the second-line treatment drug has poor curative effect, so the improvement of the treatment is always a clinically intractable problem, and the exploration of a new drug is urgently neededAnd (4) treating. Mitoxantrone liposomes in a climbing study (HE 071-01) at 12mg/m ² And 14mg/m ² Group subjects with advanced soft tissue sarcoma have shown some efficacy. The mitoxantrone liposome is used by the inventor of the invention, and the drug has the functions of slow release, targeting, toxicity reduction and synergy after entering a human body through intravenous infusion, so that the dosage is higher than that of a common injection, and the mitoxantrone liposome is single-drug therapy, thereby improving the effectiveness and reducing the occurrence probability of adverse reactions.

The study of the present invention shows that of the evaluable subjects, osteosarcoma in 3 subjects (2 subjects receiving 2-cycle treatment and 1 subject receiving 1-cycle treatment) and 1 subject was evaluated as SD (receiving 1-cycle treatment) and DCR 33.3% (1/3).

In soft tissue sarcoma, of 12 subjects (1 treatment cycle 6, 2 treatment cycle 5, 2 treatment cycle 4, 1 treatment cycle 3, 6 treatment cycle 2), 8 subjects were evaluated for SD (1 treatment cycle 6, 2 treatment cycle 5, 2 treatment cycle 4, 1 treatment cycle 3, 2 treatment cycle 2; wherein 3 subjects had decreased tumor volume), DCR 66.7% (8/12).

The mitoxantrone liposome has good curative effect on patients with osteosarcoma and soft tissue sarcoma. The 9 subjects evaluated as SD continued to take the medication, and a better therapeutic effect was obtained.

The embodiments of the present invention have been described above by way of example. It should be understood that the scope of the present invention is not limited to the above-described embodiments. Any modification, equivalent replacement, improvement or the like made by those skilled in the art within the spirit and principle of the present invention should be included in the protection scope of the claims of the present application.

Claims (10)

1. Use of mitoxantrone hydrochloride liposome in preparation of medicine for treating urothelial cancer, breast cancer, and bone and soft tissue sarcoma is provided.

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

3. The use of claim 1 or 2, wherein the urothelial cancer is locally advanced or metastatic urothelial cancer; more preferably, the urothelial cancer is locally advanced or metastatic urothelial cancer that has failed a platinum-containing chemotherapy regimen and/or a PD-1 inhibitor, or locally advanced or metastatic urothelial cancer that has failed a platinum-containing chemotherapy regimen but has been rejected for treatment with a PD-1 inhibitor, or locally advanced or metastatic urothelial cancer 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 endocrine therapy or endocrine therapy resistant, or HER-2 negative breast cancer that fails anthracycline and/or taxane therapy; preferably, the HER-2 negative breast cancer comprises immunohistochemical HER-2 0 or 1+ and immunohistochemical HER-2 + is negative breast cancer determined by in situ hybridization;

preferably, the bone and soft tissue sarcoma is an advanced bone and soft tissue sarcoma, further preferably a metastatic or locally advanced bone and soft tissue sarcoma that has failed at least one line of treatment.

4. The use according to claim 1 or 2, wherein the medicament is in the form of injection, including liquid injection, injectable powder, injectable tablet, etc.;

preferably, the medicament is a liquid injection;

preferably, the medicament contains the active ingredient in an amount of 0.5 to 5mg/ml, preferably 1 to 2mg/ml, more preferably 1mg/ml, calculated as mitoxantrone.

5. A method of treating urothelial cancer by administering to a patient having urothelial cancer a therapeutically effective amount of mitoxantrone hydrochloride liposomes; preferably, the urothelial cancer is locally advanced or metastatic urothelial cancer, more preferably the urothelial cancer is locally advanced or metastatic urothelial cancer that has failed a platinum-containing chemotherapy regimen and/or a PD-1 inhibitor, or locally advanced or metastatic urothelial cancer that has failed a platinum-containing chemotherapy regimen but has been rejected for treatment with a PD-1 inhibitor, or locally advanced or metastatic urothelial cancer that is intolerant to cisplatin.

6. A method of treating breast cancer by administering to a breast cancer patient a therapeutically effective amount of mitoxantrone hydrochloride liposomes; 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 suitably resistant to endocrine therapy or endocrine therapy; alternatively, HER-2 negative breast cancer that has failed anthracycline and/or taxane-based drug therapy; preferably, the HER-2 negative breast cancer comprises immunohistochemical HER-2 0 or 1+ and immunohistochemical HER-2 + is a breast cancer that is confirmed to be negative by in situ hybridization.

7. A method of treating a bone and soft tissue sarcoma by administering to a patient having a bone and soft tissue sarcoma a therapeutically effective amount of mitoxantrone hydrochloride liposomes; preferably, the bone and soft tissue sarcoma is advanced bone and soft tissue sarcoma, further preferably metastatic or locally advanced bone and soft tissue sarcoma that failed at least first line therapy.

8. The method of any one of claims 5-7, wherein said mode of administration is intravenous administration; preferably, the dripping administration time of the liposome pharmaceutical preparation is 30min-120min, preferably 60min-120min, and further preferably 60 +/-15 min for each intravenous administration;

preferably, the administration cycle is once every 4 or 3 weeks, preferably once every 3 weeks;

preferably, the therapeutically effective amount is 8-30mg/m, calculated as mitoxantrone ² Preferably 12 to 20mg/m ² More preferably20mg/m ² 。

9. Mitoxantrone hydrochloride liposome for treating urothelial carcinoma, breast cancer, and sarcoma of bone and soft tissue, which is characterized in that: administering a therapeutically effective amount of mitoxantrone hydrochloride liposomes to a patient with urothelial cancer, breast cancer, bone and soft tissue sarcoma; the therapeutically effective amount is 8-30mg/m, calculated as mitoxantrone ² Preferably 12 to 20mg/m ² More preferably 20mg/m ² ；

Preferably, the administration is intravenous administration; preferably, the dripping administration time of the liposome pharmaceutical preparation is 30min-120min, preferably 60min-120min, and further preferably 60 +/-15 min for each intravenous administration;

preferably, the administration cycle is once every 4 or 3 weeks, preferably once every 3 weeks.

10. The use, method or liposome of any one of claims 1-9, wherein the mitoxantrone hydrochloride liposome has one or more of the following properties: (i) The mitoxantrone hydrochloride liposome has a particle size of about 30-80nm, e.g., about 35-75nm, about 40-70nm, about 40-60nm or about 60nm;

(ii) Mitoxantrone hydrochloride forms a poorly soluble precipitate with multivalent counter ions (e.g., sulfate, citrate, or phosphate) within the liposomes;

(iii) The phospholipid bilayer of the mitoxantrone hydrochloride liposome contains phospholipids having a phase transition temperature (Tm) above body temperature, such that the liposome has a phase transition temperature above body temperature, e.g., the phospholipids are selected from the group consisting of hydrogenated soy lecithin, phosphatidylcholine, hydrogenated egg yolk lecithin, dipalmitoyl lecithin, distearoyl lecithin, or any combination thereof;

(iv) The phospholipid bilayer in the mitoxantrone hydrochloride liposome contains hydrogenated soybean lecithin, cholesterol and distearoyl phosphatidyl ethanolamine modified by polyethylene glycol 2000 (DSPE-PEG 2000);

(iiiv) the phospholipid bilayer of the mitoxantrone hydrochloride liposome comprises hydrogenated soy lecithin, cholesterol, and polyethylene glycol 2000 modified distearoylphosphatidylethanolamine at a mass ratio of about 3;

preferably, the mitoxantrone hydrochloride liposomes have a particle size of about 30-80nm and comprise: 1) Mitoxantrone, an active ingredient, which forms a poorly soluble precipitate with multivalent counter ions in liposomes, 2) phospholipid bilayers comprising phospholipids with a phase transition temperature (Tm) above body temperature; the phospholipid with Tm higher than body temperature is phosphatidyl choline, hydrogenated soybean lecithin, hydrogenated yolk lecithin, dipalmitin lecithin or distearoyl lecithin or any combination thereof;

preferably, the mitoxantrone hydrochloride liposome has a particle size of about 35-75nm, preferably 40-70nm, more preferably 40-60nm, and particularly preferably 60nm;

or preferably, the phospholipid bilayer contains hydrogenated soybean lecithin, cholesterol and polyethylene glycol 2000 modified distearoyl phosphatidyl ethanolamine, the mass ratio is 3;

or preferably, the phospholipid bilayer of the liposome comprises hydrogenated soybean lecithin, cholesterol and polyethylene glycol 2000 modified distearoyl phosphatidyl ethanolamine, the mass ratio is 3: chol: DSPE-PEG2000: the weight ratio of mitoxantrone is 9.58.