Use of mitoxantrone liposome for treating non-Hodgkin's lymphoma

Abstract

Disclosed is a use of a liposomal pharmaceutical preparation of mitoxantrone in the preparation of a medicament for treating lymphoma, wherein the lymphoma is preferably non-Hodgkin's lymphoma, further preferably aggressive non-Hodgkin's lymphoma, more preferably diffuse large B-cell lymphoma or peripheral T-cell lymphoma, and more further preferably relapsed or refractory diffuse large B-cell lymphoma or peripheral T-cell lymphoma. The mitoxantrone liposomes are used as single anti-tumor therapeutic agent without being combined with other anti-tumor agents.

Claims

1. A method for treating lymphoma, comprising administering to a subject in need thereof a liposomal pharmaceutical preparation of mitoxantrone as a single anti-tumor therapeutic agent, wherein: the liposomal pharmaceutical preparation of mitoxantrone comprises mitoxantrone liposomes, the mitoxantrone liposomes have a particle size of about 30-80 nm, and comprise mitoxantrone as an active ingredient, wherein mitoxantrone and a multivalent counter ion within the liposomes form a poorly soluble precipitate, the phospholipid bilayer of the liposomes comprises a phospholipid with a phase transition temperature (Tm) higher than body temperature, so that the phase transition temperature of liposomes is higher than the body temperature; and the lymphoma is relapsed or refractory diffuse large B-cell lymphoma or peripheral T-cell lymphoma.

2. The method of claim 1, wherein the phospholipid with a Tm higher than body temperature is selected from phosphatidylcholine, hydrogenated soybean phosphatidylcholine, hydrogenated egg yolk phosphatidylcholine, dipalmitoyl phosphatidylcholine, distearoyl phosphatidylcholine and any combination thereof; and/or the liposomes have a particle size of about 40-70 nm.

3. The method of claim 1, wherein the phospholipid bilayer contains hydrogenated soybean phosphatidylcholine, cholesterol, and polyethylene glycol 2000-modified distearoylphosphatidylethanolamine at a mass ratio of 3:1:1, and/or the liposomes have a particle size of about 60 nm, and/or the counter ion is sulfate ion.

4. The method of claim 1, wherein the subject has diffuse large B-cell lymphoma and at least has received a first-line and/or second-line treatment, or the subject has peripheral T-cell lymphoma and at least has received a first-line treatment.

5. The method of claim 1, wherein the liposomal pharmaceutical preparation of mitoxantrone is administered once per 3-6 weeks; and/or the liposomal pharmaceutical preparation of mitoxantrone is administered to the subject 2, 3, 4 or 5 times; and/or the liposomal pharmaceutical preparation of mitoxantrone is administrated to the subject at a dosage of 14-24 mg/m.sup.2.

6. The method of claim 1, wherein the liposomal pharmaceutical preparation is administered to the subject via intravenous drip, and the administration period via drip is 40-80 min.

7. The method of claim 1, wherein the total dosage of the liposomal pharmaceutical preparation administered to the subject is not more than 200 mg/m.sup.2.

8. The method of claim 1, wherein the route of administration is selected from oral administration, local administration, subcutaneous injection, intramuscular injection, intravenous drip and intravenous bolus injection.

9. The method of claim 1, wherein the route of administration is intravenous drip, and the method comprises dissolving the mitoxantrone liposomes in sodium chloride injection or glucose injection, and then administering to the subject; wherein the concentration of sodium chloride injection is 0.5%-1%; the concentration of glucose injection is 1%-10%.

10. The method of claim 1, comprising formulating the mitoxantrone liposomes into a solution that can be directly infused before administering to the subject, wherein the concentration of the formulated solution of liposomes that can be directly infused is 0.05 mg/ml-0.5 mg/ml.

11. The method of claim 1, comprising administering to the subject the liposomal pharmaceutical preparation of mitoxantrone 3 or more times for treatment, and the method achieves a higher overall remission rate than a histone deacetylase inhibitor used alone.

12. The method of claim 1, comprising administering to the subject the liposomal pharmaceutical preparation of mitoxantrone 3 or more times for treatment, and the overall remission rate of the method is at least 30%.

13. The method of claim 2, wherein the liposomes have a particle size of 60 nm.

14. The method of claim 5, wherein the liposomal pharmaceutical preparation of mitoxantrone is administered once per 4 weeks.

15. The method of claim 6, wherein the administration period via drip is 60 min.

Description

EXAMPLES

Example 1 Preparation of Mitoxantrone Liposomes

(1) HSPC, Chol and DSPE-PEG2000 were weighed at a mass ratio of 3:1:1 and dissolved in 95% ethanol to obtain a clear solution. The ethanol solution of phospholipids was mixed with 300 mM ammonium sulfate solution, shaken and hydrated for 1 h at 60-65° C. to obtain heterogeneous multilamellar liposomes. Then a Microfluidizer was used to reduce the particle size of liposomes. The obtained sample was diluted 200 times with a 0.9% NaCl solution, and then detected with NanoZS. The average particle size of the particles was about 60 nm, and the main peak was between 40 nm and 60 nm. Afterwards, an ultrafiltration device was used to remove the ammonium sulfate in the outer phase of the blank liposomes, and the outer phase was replaced with 290 mM sucrose and 10 mM glycine to form a transmembrane ammonium sulfate gradient. Mitoxantrone hydrochloride solution (10 mg/mL) was added to the blank liposomes at a lipid:drug ratio of 16:1, and the drug was loaded at 60-65° C. After incubating for about 1 h, gel exclusion chromatography was used to prove that the encapsulation efficiency was about 100%. The weight ratio of HSPC:Chol:DSPE-PEG2000:mitoxantrone was 9.58:3.19:3.19:1, and the osmotic pressure of the sucrose-glycine solution was close to the physiological value.

Example 2 Investigation on the Stability of Mitoxantrone Liposomes

(2) The mitoxantrone liposomes prepared by the above-mentioned method were prepared into two samples at the concentration of 0.1 mg/ml and 0.2 mg/ml by using 5% glucose injection and 0.9% sodium chloride injection (normal saline) as diluents and using glass infusion bottles as containers, respectively. The parameters such as pH of the solution, average particle size of liposomes, concentration of encapsulated drug, phospholipids, lysolecithin, related substances, and content were examined at 30° C., at 30° C. in the dark and under refrigerated condition at 2-8° C. at six time points of 0 h, 2 h, 4 h, 6 h, 8 h and 24 h, respectively. The experimental results are shown in Table 1-4 below.

(3) TABLE-US-00002 TABLE 1 Stability of mitoxantrone liposomes diluted with 5% glucose injection, glass infusion bottle, 0.1 mg/mL Concentration Related of substance Average encapsulated (%) Time particle drug Phospholipids Mitoxantrone Maximum Con- point size (mg/ (mg/mL) Lysolecithin peak single Content dition (h) pH (nm) mL) Cholesterol HSPC.sub.C16-18 HSPC.sub.C18-18 (mg/mL) purity impurity (mg/mL) 30° 0 6.47 63.1 0.085 0.279 0.192 0.669 not 99.82 0.06 0.091 C. detected 2 6.45 63.6 0.087 0.286 0.197 0.678 ND 99.85 0.05 0.091 4 6.46 63.3 0.087 0.289 0.199 0.671 ND 99.86 0.06 0.090 6 6.46 63.4 0.086 0.289 0.199 0.670 ND 99.89 0.04 0.089 8 6.47 63.5 0.086 0.291 0.200 0.674 ND 99.90 0.04 0.093 24 6.48 63.5 0.088 0.287 0.191 0.661 not 99.94 0.03 0.090 detected Average 6.5 63 0.087 0.287 0.196 0.670 N/A 99.88 0.05 0.091 value RSD 0.2 0.3 1.1 1.4 2.0 0.8 N/A 0.1 26.8 1.3 (%) 30° 0 6.46 62.9 0.086 0.286 0.197 0.669 not 99.87 0.05 0.091 C. in detected the 2 6.47 63.4 0.087 0.289 0.193 0.675 ND 99.88 0.05 0.091 dark 4 6.47 63.9 0.086 0.292 0.200 0.686 ND 99.89 0.04 0.091 6 6.45 62.9 0.088 0.288 0.197 0.665 ND 99.90 0.04 0.092 8 6.46 63.6 0.086 0.288 0.200 0.678 ND 99.87 0.05 0.091 24 6.46 64.0 0.088 0.283 0.194 0.675 not 99.94 0.03 0.091 detected Average 6.5 63 0.087 0.288 0.197 0.675 N/A 99.89 0.04 0.091 value RSD 0.1 0.7 1.3 1.0 1.5 1.1 N/A 0.1 19.6 0.5 (%) Refrig- 0 6.46 63.8 0.084 0.287 0.192 0.687 not 99.66 0.06 0.091 erated detected at 2-8° 2 6.47 62.9 0.088 0.291 0.192 0.683 ND 99.81 0.06 0.091 C. 4 6.47 63.4 0.087 0.287 0.195 0.672 ND 99.81 0.06 0.091 6 6.47 63.7 0.088 0.297 0.198 0.682 ND 99.76 0.07 0.091 8 6.46 62.8 0.087 0.282 0.187 0.681 ND 99.77 0.07 0.092 24 6.46 63.6 0.087 0.283 0.193 0.686 not 99.84 0.06 0.090 detected Average 6.5 63 0.087 0.288 0.193 0.682 N/A 99.77 0.06 0.091 value RSD 0.1 0.7 1.7 1.8 2.0 0.8 N/A 0.1 8.2 0.8 (%) N/A: Not applicable; ND: Not detected

(4) TABLE-US-00003 TABLE 2 Stability of mitoxantrone liposomes diluted with 0.9% sodium chloride injection, glass infusion bottle, 0.1 mg/mL Concentration Related of substance Average encapsulated (%) Time particle drug Phospholipids Mitoxantrone Maximum Con- point size (mg/ (mg/mL) Lysolecithin peak single Content dition (h) pH (nm) mL) Cholesterol HSPC.sub.C16-18 HSPC.sub.C18-18 (mg/mL) purity impurity (mg/mL) 30° 0 6.56 62.0 0.088 0.284 0.196 0.694 not 99.82 0.11 0.093 C. detected 2 6.57 62.3 0.088 0.287 0.195 0.711 ND 99.80 0.10 0.093 4 6.56 63.4 0.087 0.291 0.198 0.703 ND 99.84 0.11 0.094 6 6.58 63.8 0.088 0.287 0.192 0.713 ND 99.85 0.11 0.094 8 6.57 63.2 0.088 0.289 0.201 0.697 ND 99.82 0.11 0.093 24 6.55 65.4 0.089 0.287 0.201 0.693 not 99.84 0.10 0.093 detected Average 6.6 63 0.088 0.288 0.197 0.702 N/A 99.83 0.11 0.093 value RSD 0.2 1.9 0.7 0.9 1.7 1.2 N/A 0.1 4.2 0.7 (%) 30° 0 6.58 61.9 0.088 0.282 0.197 0.681 not 99.84 0.06 0.094 C. detected in the 2 6.56 62.1 0.087 0.282 0.200 0.694 ND 99.85 0.06 0.094 dark 4 6.56 63.5 0.089 0.288 0.190 0.702 ND 99.82 0.07 0.093 6 6.59 63.4 0.088 0.289 0.194 0.701 ND 99.85 0.06 0.094 8 6.57 64.3 0.088 0.283 0.192 0.688 ND 99.79 0.10 0.093 24 6.58 66.1 0.088 0.290 0.196 0.700 not 99.81 0.09 0.094 detected Average 6.6 64 0.088 0.286 0.195 0.694 N/A 99.83 0.07 0.093 value RSD 0.2 2.4 0.7 1.3 1.8 1.2 N/A 0.1 24.2 0.7 (%) Refrig- 0 6.59 61.6 0.089 0.280 0.195 0.694 not 99.83 0.06 0.093 erated detected at 2-8° 2 6.61 62.7 0.089 0.288 0.204 0.717 ND 99.82 0.07 0.095 C. 4 6.58 62.9 0.087 0.283 0.198 0.702 ND 99.82 0.07 0.094 6 6.58 63.9 0.088 0.283 0.198 0.690 ND 99.83 0.08 0.093 8 6.61 64.0 0.088 0.283 0.196 0.684 ND 99.76 0.12 0.093 24 6.59 63.0 0.088 0.284 0.194 0.680 not 99.82 0.08 0.093 Average 6.6 63 0.088 0.284 0.198 0.694 N/A 99.81 0.08 0.093 value RSD 0.2 1.4 0.8 0.9 1.8 1.9 N/A 0.1 24.1 0.8 (%) N/A: Not applicable; ND: Not detected

(5) TABLE-US-00004 TABLE 3 Stability of mitoxantrone liposomes diluted with 5% glucose injection, glass infusion bottle, 0.2 mg/mL Concentration Related of substance Average encapsulated (%) Time particle drug Phospholipids Mitoxantrone Maximum Con- point size (mg/ (mg/mL) Lysolecithin peak single Content dition (h) pH (nm) mL) Cholesterol HSPC.sub.C16-18 HSPC.sub.C18-18 (mg/mL) purity impurity (mg/mL) 30° 0 6.57 63.0 0.182 0.601 0.404 1.445 not 99.87 0.10 0.186 C. detected 2 6.57 63.0 0.181 0.603 0.396 1.411 ND 99.86 0.10 0.189 4 6.56 63.1 0.179 0.595 0.395 1.374 ND 99.85 0.11 0.189 6 6.59 63.2 0.183 0.586 0.404 1.404 ND 99.85 0.10 0.188 8 6.59 63.1 0.182 0.592 0.397 1.388 ND 99.85 0.11 0.188 24 6.58 64.0 0.179 0.581 0.401 1.371 not 99.86 0.10 0.186 detected Average 6.6 63 0.181 0.593 0.399 1.399 N/A 99.86 0.10 0.188 value RSD 0.2 0.6 1.0 1.4 1.0 2.0 N/A 0.1 4.9 0.7 (%) 30° 0 6.55 63.3 0.179 0.605 0.398 1.405 not 99.85 0.10 0.186 C. detected in 2 6.56 63.6 0.180 0.589 0.400 1.392 ND 99.82 0.11 0.186 the 4 6.56 63.0 0.181 0.591 0.392 1.396 ND 99.87 0.09 0.185 dark 6 6.57 62.7 0.181 0.599 0.381 1.387 ND 99.84 0.11 0.185 8 6.57 63.4 0.181 0.595 0.393 1.414 ND 99.83 0.11 0.187 24 6.56 63.3 0.179 0.579 0.398 1.347 not 99.84 0.11 0.186 detected Average 6.6 63 0.180 0.593 0.393 1.390 N/A 99.84 0.10 0.186 value RSD 0.1 0.5 0.5 1.5 1.8 1.6 N/A 0.1 6.9 0.4 (%) Refrig- 0 6.56 63.5 0.182 0.589 0.407 1.394 not 99.85 0.10 0.188 erated detected at 2-8° 2 6.56 62.8 0.181 0.594 0.398 1.386 ND 99.85 0.10 0.190 C. 4 6.55 62.8 0.180 0.598 0.395 1.417 ND 99.84 0.11 0.189 6 6.57 63.1 0.182 0.580 0.396 1.388 ND 99.85 0.10 0.189 8 6.56 63.6 0.180 0.585 0.389 1.395 ND 99.78 0.11 0.188 24 6.56 63.2 0.174 0.580 0.387 1.375 not 99.80 0.10 0.185 detected Average 6.6 63 0.180 0.588 0.395 1.392 N/A 99.83 0.10 0.188 value RSD 0.1 0.5 1.7 1.2 1.8 1.0 N/A 0.1 2.3 0.9 (%) N/A: Not applicable; ND: Not detected

(6) TABLE-US-00005 TABLE 4 Stability of mitoxantrone liposomes diluted with 0.9% sodium chloride injection, glass infusion bottle, 0.2 mg/mL Concentration Related of substance Average encapsulated (%) Time particle drug Phospholipids Mitoxantrone Maximum Con- point size (mg/ (mg/mL) Lysolecithin peak single Content dition (h) pH (nm) mL) Cholesterol HSPC.sub.C16-18 HSPC.sub.C18-18 (mg/mL) purity impurity (mg/mL) 30° 0 6.58 61.8 0.183 0.568 0.407 1.394 not 99.80 0.06 0.189 C. detected 2 6.57 63.1 0.182 0.574 0.411 1.401 ND 99.85 0.06 0.188 4 6.57 62.9 0.178 0.569 0.404 1.391 ND 99.80 0.08 0.190 6 6.59 62.6 0.182 0.569 0.405 1.385 ND 99.78 0.12 0.191 8 6.57 63.3 0.182 0.574 0.412 1.377 ND 99.87 0.08 0.189 24 6.57 65.3 0.179 0.577 0.392 1.419 not 99.81 0.08 0.191 detected Average 6.6 63 0.181 0.572 0.405 1.395 N/A 99.82 0.08 0.190 value RSD 0.1 1.9 1.0 0.6 1.8 1.0 N/A 0.1 29.9 0.6 (%) 30° 0 6.59 62.5 0.178 0.564 0.409 1.385 not 99.85 0.10 0.185 C. 2 6.58 63.4 0.180 0.568 0.401 1.382 ND 99.84 0.11 0.186 in 4 6.56 63.3 0.181 0.570 0.404 1.435 ND 99.84 0.11 0.188 the 6 6.55 63.2 0.172 0.572 0.411 1.404 ND 99.83 0.11 0.186 dark 8 6.57 63.3 0.173 0.570 0.402 1.383 ND 99.83 0.11 0.189 24 6.58 65.9 0.175 0.579 0.400 1.393 not 99.81 0.11 0.189 detected Average 6.6 64 0.176 0.571 0.404 1.397 N/A 99.83 0.11 0.187 value RSD 0.2 1.8 1.9 0.9 1.1 1.4 N/A 0.1 2.5 0.8 (%) Refrig- 0 6.55 62.9 0.183 0.579 0.408 1.386 not 99.85 0.06 0.186 erated detected at 2-8° 2 6.54 62.1 0.181 0.574 0.421 1.404 ND 99.89 0.06 0.188 C. 4 6.59 62.3 0.180 0.578 0.406 1.425 ND 99.79 0.11 0.192 6 6.57 62.0 0.182 0.574 0.405 1.399 ND 99.83 0.11 0.190 8 6.58 64.6 0.179 0.573 0.401 1.418 ND 99.75 0.19 0.189 24 6..56 62.5 0.179 0.588 0.400 1.416 not 99.84 0.06 0.190 detected Average 6.6 63 0.181 0.578 0.407 1.408 N/A 99.82 0.10 0.189 value RSD 0.3 1.6 0.9 1.0 1.9 1.0 N/A 0.1 54.0 0.9 (%) N/A: Not applicable; ND: Not detected

(7) The above results show that, under the same concentration and the same storage condition, by using glucose injection and sodium chloride injection as diluents, the parameters such as pH of the obtained solutions, concentration of encapsulated drug, phospholipids, lysolecithin, related substances, and content over time are not significantly different, while the average particle sizes of mitoxantrone liposomes are significantly different. When 0.9% sodium chloride injection is used as the diluent, the average particle size of mitoxantrone liposomes from 0 h to 24 h has an obvious increasing trend, but when 0.5% glucose injection is used as the diluent, the average particle size of the liposomal drug from 0 h to 24 h has little change.

(8) In addition, under the same diluent and the same storage condition, there are no significant differences in parameters of mitoxantrone liposomes with the drug concentrations of 0.1 mg/ml and 0.2 mg/ml, such as pH of the solutions, average particle size, concentration of encapsulated drug, phospholipids, lysolecithin, related substances, and content over time.

Example 3 Example of Administration Method

(9) The mitoxantrone liposomes (specification: 10 mg/10 ml/vial) obtained as above was dissolved in 250 ml of 5% glucose injection, and the drip period was 60 min. Intravenous drip was carried out on forearm at one side, and the semi-recumbent position was adopted during administration. The whole process of intravenous administration was supervised by clinicians.

Example 4 Treatment of DLBCL by Using Mitoxantrone Liposomes Alone Inclusion Criteria

(10) A phase II clinical study of mitoxantrone liposomes for treating DLBCL and peripheral T/NK cell lymphoma was conducted. The inclusion criteria for this study is as follows:

(11) 1) voluntarily underwent the test and signed informed consent;

(12) 2) aged 18 to 75 years, with no gender limitations;

(13) 3) ECOG score 0-2 grade;

(14) 4) expected survival time ≥3 months;

(15) 5) non-Hodgkin's lymphomas of diffuse large B-cells and peripheral T/NK cells are confirmed by histopathology, wherein peripheral T/NK cell lymphomas are limited to the following types: peripheral T-cell lymphoma (non-specific type), angioimmunoblastic T-cell lymphoma, ALK+systemic anaplastic large T-cell lymphoma, ALK− systemic anaplastic large T-cell lymphoma, extranodal NK/T-cell lymphoma, nasal-type, enteropathy-associated T-cell lymphoma, primary hepatosplenic γδ T-cell lymphoma, and subcutaneous panniculitis-like T-cell lymphoma;

(16) 6) patients who have experienced at least one systemic treatment failure or recurrence in the past and cannot undergo autologous peripheral blood hematopoietic stem cell transplantation;

(17) 7) there is at least one measurable lesion, and the long diameter of a single measurable lesion is ≥1.5 cm;

(18) 8) the target tumor is at least 4 weeks apart from the last chemotherapy, radiotherapy, biological therapy, stem cell transplantation or other experimental drug treatment;

(19) 9) subjects of childbearing age agree to take effective contraceptive measures during the trial; blood pregnancy test results of females are negative (except for postmenopausal or surgery induced infertility);

(20) 10) the level of organ function is basically normal.

(21) Safety Evaluation:

(22) Safety assessment includes adverse events, physical examination and other tests (blood routine, urine routine, blood biochemistry, electrocardiogram, UCG), and early withdrawal due to safety or tolerance reasons. Safety evaluation standard: NCI-CTC4.0.

(23) Effectiveness Evaluation:

(24) Efficacy evaluation method: the patients underwent a tumor evaluation during the baseline period, and the CT scan results of the tumor site should prevail. Efficacy evaluation was conducted according to the standards of the International Workshop Criteria (IWC) for Malignant Lymphoma (2007).

(25) Effectiveness Analysis:

(26) In this study, we analyzed patients with DLBCL. Subjects received an average of 3.0±2.0 cycles of drugs for treatment. Mitoxantrone liposomes were administered at a dosage of 16 mg/m.sup.2 or 20 mg/m.sup.2 once per 28 days. Of the 19 DLBCL patients with evaluable efficacy, 10 achieved CR or PR, with an ORR of 52.6%.

(27) The ORR of the combination chemotherapy regimen is about 60%-70% (ICE regimen: the combination of three drugs, i.e. ifosfamide, carboplatin, and etoposide has an ORR of 66%; the combination of five drugs, i.e. rituximab, etoposide, methylprednisolone, cytarabine and cisplatin has an ORR of 73%) (Treatment challenges in the management of relapsed or refractory non-Hodgkin's lymphoma—novel and emerging therapies, Journal of Cancer Management and Research, Mark P Chao et al., 2013, Vol. 5, pp. 251-269), which is equivalent to the efficacy of mitoxantrone liposomes used alone in this Example, and there is no statistical difference.

(28) The efficacy of mitoxantrone liposomes clinically used alone for treating DLBCL is shown in Table 5 below in details.

(29) TABLE-US-00006 TABLE 5 Efficacy of mitoxantrone liposomes clinically used alone for treating DLBCL Number of Dosage administration Previous history of Best No. (mg/m.sup.2) cycles chemotherapy efficacy 1 20 1 CHOP × 3 PR DICE × 2 Gemox × 4 2 20 5 CHOP × UK PR 3 16 6 HLX01/R-CHOP × PR 6 4 16 2 HLX01/ PD R-CHOP × 6 EP × 2 etoposide × UK 5 16 4 CHOP × 4 PR 6 20 2 R-CHOP + SD ibrutinib × 5 ICE × 2 7 16 2 R-EPOCH × 6 SD DICE × 6 8 16 2 R-CHOP × 6 PD FC × UK 9 16 2 R-CHOP × 1 PD CHOP × 2 ICE × 2 GCP × 1 CHOP + Mesna × 1 10 16 2 R-MTX × 2 PD R-CHOP × 6 GEMOX + TNF × 4 FC × 1 11 16 2 CHOP × 6 PR CHOP × 4 ECOP × 2 CTP × 1 EPP × 4 EP × 1 12 16 1 OB × 3 PD CHOP × 4 ABVD × 2 13 16 6 R-CHOP × 6 CR 14 16 2 CDOP × 7 PR GDP × 1 DICE × 4 15 16 1 R-CHOP × 6 PD DICE + Chidamide × 3 16 16 5 R-CHOP × 4 PR R-DA-EPOCH × 4 GDE × 4 R-GDP × 1 R-GDP × 4 R × UK R-GDE × 2 17 16 6 CHOP × 4 PR DICE × 5 R-NP × 3 18 16 1 R-CHOP × 4 PD R-EPOCH × 2 19 16 6 CHOP × 6 PR Gemox × 3 Note: the meanings of the above abbreviations are as follows: CR: complete remission, defined as the disappearance of all evidences of disease. PR: partial remission, defined as measurable lesion shrinkage without new lesion. PD: progression of disease, defined as the appearance of any new lesion, or an increase of ≥50% compared with the lowest point of the original lesion. SD: stable disease, defined as not belonging to any of the situations. Overall remission rate (ORR) = (CR + PR)/total number of evaluable cases * 100%

(30) CR: complete remission, defined as the disappearance of all evidences of disease.

(31) PR: partial remission, defined as measureable lesion shrinkage without new lesion.

(32) PD: progression of disease, defined as the appearance of any new lesion, or an increase of ≥50% compared with the lowest point of the original lesion.

(33) SD: stable disease, defined as not belonging to any of the situations.
Overall remission rate (ORR)=(CR+PR)/total number of evaluable cases*100%

Example 5 Analysis of Adverse Reactions of Mitoxantrone Liposomes Used Alone for Treating DLBCL

(34) Safety Analysis:

(35) The adverse reactions occurred in the above-mentioned DLBCL subjects were analyzed, and the most frequent adverse reaction was hematological toxicity.

(36) As for the hematological toxicity, 31.4% of subjects developed grade 3 or higher leukopenia, 28.6% of the subjects developed grade 3 or higher neutropenia, and 5.7% of the subjects developed grade 3 or higher thrombocytopenia. This is much lower than the hematological toxicity disclosed in the reference analyzed in the background art (Phase II study of the irinotecan (CPT-11), mitoxantrone and dexamethasone regimen in elderly patients with relapsed or refractory peripheral T-cell lymphoma, Journal of Cancer Sci, Nozomi Niitsu et al., 2007, Vol. 98, pp. 109-112) (in this study, 18 cases (60%) developed grades 3-4 hematological toxicity. 8 cases (27%) developed grade 4 neutropenia despite the use of leucocyte increasing agents (G-CSF). 5 cases (16.7%) developed grades 3-4 thrombocytopenia).

(37) As for the non-hematological toxicity, 5.7% of subjects developed grade 3 or higher hypoalbuminemia, 5.7% of subjects developed grade 3 or higher lung infections, 2.9% of subjects developed grade 3 or higher intestinal obstruction, and 2.9% of subjects developed grade 3 loss of appetite.

Example 6 Treatment of PTCL with Mitoxantrone Liposomes Used Alone

(38) The inclusion criteria, administration method, safety and efficacy evaluation are the same as in Example 1.

(39) The mitoxantrone liposomes developed by our company is considered as a second-line treatment for treating PTCL. The ORR of this product in clinical study is 52.9%, which is much higher than those of the HDAC inhibitors including pralatrexate, romidepsin, belinostat, chidamide and the like currently recommended as second-line treatments (HDAC inhibitors are oral preparations and administered once a day. All of the ORR are lower than 30%). Patients who achieve remission in a short period of time can undergo bone marrow transplantation in time.

(40) Mitoxantrone liposomes are injections, which are used at the dosages of 14, 16, 20 mg/m.sup.2 or 24 mg/m.sup.2, and are administered once per 28 days. The subjects in this study received an average of 3.6±1.8 cycles of treatment.

(41) The efficacy of mitoxantrone liposomes clinically used alone for treating PTCL is shown in Table 6 below in details.

(42) TABLE-US-00007 TABLE 6 Efficacy of mitoxantrone liposomes clinically used alone for treating PTCL Number of No. of the Dosage administration Previous history of Best patient (mg/m.sup.2) cycles chemotherapy efficacy 1 14 3 CHOEP × 3 CR CHOP × 4 DICE × 4 2 24 2 Gemcitabine × 4 PD CHOP × 4, GDP × 2 3 20 6 CHOP + PR LASPAR × 3 GEMOX × 2 Chidamide × 2 GEMOX × 2 4 20 5 CHOPE × 5 PR 5 20 3 CHOPE/GDP × 6, PR Pralatrexate × 5 6 20 2 CHOP × 4, NA COP + (withdrawal) Lenalidomide × 1 D-RPOCH × 1 DA-EPOCH × 1 7 20 6 CHOP × 6 PR 8 16 6 CHOP × 6 PR DICE × 3 DHAP × 1 9 16 2 CHOP × 1 NA EPOCH × 6 (withdrawal) GDP × 6 DICE + Thalidomide × 2 Pralatrexate × 9 10 16 2 CHOPE/GDP × 4 SD DICE × 1 CE × 4 CTX + VP-16 × 5 EPOCH × 2 11 16 3 CHOP × 7 SD 12 16 2 CHOPE × 4 SD 13 16 6 CHOP × 7 PR 14 16 3 CHOP × 4 PR ECHOP × 2 15 16 5 Asparaginase + SD CHOP × 2 CHOP × 1 Pegaspargase + CHO × 3 CHOP × 2 16 16 4 CHOP × 1 PR R-mini-chop × 1 CHOP × l R-CHOP × 4 17 16 1 AEOPP × 1 PD CHOP × 4

(43) Note: among them, the meanings of CR, PR, PD, SD, ORR, efficacy evaluation method and efficacy evaluation standard are as described in the “Note” part of Table 5.

Example 7 Analysis of Adverse Reactions of Mitoxantrone Liposomes Used Alone for Treating PTCL

(44) The adverse reactions occurred in 15 patients with PTCL were analyzed, and the most frequent adverse reaction was hematological toxicity.

(45) As for the hematological toxicity, 9 subjects (60.0%) developed grade 3 leukopenia, 3 subjects (20%) developed grade 4 leukopenia; 7 subjects (46.7%) developed grade 3 neutropenia, 1 subject (6.7%) developed grade 4 neutropenia; 1 subject developed grade 3 thrombocytopenia; 1 subject developed grade 3 reduction in hemoglobin; and 1 subject developed grade 3 erythropenia.

(46) As for the non-hematological toxicity, no toxicity of grade 3 or higher has occurred.

(47) By comparison, although the incidences of grades 3-4 hematological toxicity were similar, the incidences of grade 4 neutropenia and grades 3-4 thrombocytopenia were all lower than the CMD regimen reported in the previous reference.