Compositions containing antitumor drugs, and methods for preparing and using the same

Abstract

A composition comprising 7-ethyl-10-hydroxycamptothecin, as well as a method for preparing and using the same, is disclosed. The composition comprises SN-38, lipid, albumin, and Span 20.

Description

[Technical Field]

[0001] The present application relates to a composition of 7-ethyl-10-hydroxycamptothecin (SN-38) comprising SN-38, lipid, albumin, and Span 20, as well as methods for preparing and using the same. [Background technology]

[0002] SN-38 is an active metabolite of the commercially available drug irinotecan hydrochloride (CPT-11) in the body and exhibits approximately 100-1000 times greater efficacy against several tumor cells than CPT-11 (Zhang JA, Xuan T, Parmar M, et al., Development and characterization of a novel liposome-based formulation of SN-38, [J]. International journal of pharmaceutics, 2004, 270(1):93-107). SN-38 exhibits inhibitory effects against various tumor cells, including colorectal cancer, small cell lung cancer, lymphoma, breast cancer, esophageal cancer, uterine cancer, and ovarian cancer. However, the conversion efficiency of CPT-11 to SN-38 in the body is very low, at only 2-8% (Rowinsky EK, Grochow LB, Ettinger DS, et al., Phase I and pharmacological study of the novel topoisomerase I inhibitor CPT-11 administered as a 90-minute infusion every 3 weeks, [J]. Cancer research, 1994, 54(2):427-436).

[0003] Studies have shown that the closed-ring lactone structure of SN-38 is an effective component for exerting its anticancer activity. However, SN-38 with this closed-ring structure is insoluble in most biocompatible and pharmaceutically acceptable solvents, making it less druggable. These factors significantly limit the development and clinical use of SN-38. To date, no pharmaceutical formulations containing SN-38 as an active ingredient have been approved for marketing.

[0004] Therefore, there is still an urgent need to solve the problems of SN-38's solubility and druggability. To solve the solubility problem of SN-38, several studies have focused on structural modifications of SN-38. These modifications can be classified into water-soluble modifications and lipophilic modifications, the former of which can be found, for example, in International Publication No. 1995022549, and the latter of which can be found, for example, in U.S. Patent Application Publication No. 20060229359. Lipophilic modifications usually involve preparing SN-38 as liposomes. This preparation process typically involves modifying camptothecin molecules with hydrophobic molecules such as long-chain fatty acids or cholesterol, vitamin E, and lipophilic organic acids, followed by preparing camptothecin-liposome products with the modified camptothecin molecules and a certain proportion of excipients such as phospholipids and cholesterol. Chinese Patent Application Publication No. 108567742 relates to obtaining camptothecin-liposome products by improving excipients rather than hydrophobically modifying SN-38. Although the products reported above do not contain albumin, modification with lipids allows camptothecin to maintain a large proportion of its active closed ring structure and allows camptothecin to be trapped by albumin after entering the body, thereby prolonging its effectiveness.

[0005] Furthermore, due to the unique physical and chemical properties of SN-38 compared to paclitaxel drug, such as its easier crystallization in water and on positively charged surfaces, the stability and scale-up of the preparation process of SN-38 formulations have always been challenges for the druggability of SN-38.

[0006] Nanoliposomes, nanosuspensions, etc. obtained by water-soluble or lipophilic modified / liposome preparations have drawbacks such as insufficient SN-38 drug loading, complicated preparation processes, poor reproducibility after process scale-up, and unstable formulations. It remains an urgent problem to achieve cost-effective SN-38 nanoformulations with high drug loadings through a process that is easy to scale up and stable through the optimization of formulation components and processes.

[0007] Our prior application (International Application PCT / CN2021 / 102332) describes a high-drug-loading composition containing SN-38, lipids, and albumin, resulting in an SN-38 formulation suitable for drug preparation. Based on the above research, we surprisingly discovered that adding Span 20 to the composition reduces the number of high-pressure homogenization steps during preparation, substantially reduces the particle size of the formulation, increases filtration flux, and reduces raw material loss and costs. At the same time, the resulting pharmaceutical formulation exhibits improved stability, improved particle size control after disintegration, and more consistent efficacy. The contents of International Application PCT / CN2021 / 102332 are incorporated herein by reference in their entirety. Summary of the Invention

[0008] International Patent Application PCT / CN2021 / 102332 describes a composition containing SN-38, lipids, and albumin. The inventors discovered that adding Span 20 to a composition in a scaled-up process, such as a pilot-scale preparation, on a relatively large scale (e.g., 100 milligrams or more of SN-38 raw material) can reduce the number of high-pressure homogenization steps during preparation, substantially reduce the particle size of the nanoparticles in the composition, increase the filtration flux, reduce raw material loss and costs, and simultaneously control the particle size of the nanoparticles after disintegration. By further controlling the albumin content in the composition, the particle size of the nanoparticles can be controlled to approach a particle size suitable for drug preparation. The composition of the present invention also has the advantages of the composition in International Application PCT / CN2021 / 102332, including: (1) increased drug loading and encapsulation efficiency of SN-38; (2) low levels of SN-38 with an open-ring structure; (3) absence of albumin multimers, low immunogenicity, and high safety; (4) small nanoparticle size and narrow particle size distribution; (5) excellent stability (including excellent dilution stability and storage stability); (6) SN-38 exists in an amorphous and / or nanocrystalline form, which has the advantages of a high dissolution rate and high bioavailability; and (7) excellent in vivo antitumor efficacy.

[0009] In a first aspect, the present application provides a composition comprising SN-38, a lipid, albumin, and Span 20, wherein the composition comprises nanoparticles in which the albumin encapsulates at least a portion of the SN-38 and optionally at least a portion of the lipid, and wherein the lipid is selected from cholesterol, cholesterol derivatives, cholesterol analogs, and fatty acid esters, and any combination of two or more thereof.

[0010] In a second aspect, the present application provides a method for preparing a composition according to the first aspect.

[0011] In a third aspect, the present application provides a method for preparing a composition comprising SN-38, a lipid, an albumin, and Span 20.

[0012] In a fourth aspect, the present application provides a composition preparable by the method according to the third aspect.

[0013] In a fifth aspect, the present application further provides a method for preparing a composition with improved properties.

[0014] Furthermore, other aspects of the present application provide pharmaceutical compositions containing the above compositions, and uses thereof. [Brief explanation of the drawings]

[0015] [Figure 1] 1 is a typical HPLC chromatogram for measuring the content of SN-38 in the product prepared in Example 1. [Figure 2] 1 is a typical HPLC chromatogram for measuring the cholesterol content in the product prepared in Example 1. [Figure 3] 1 is a typical HPLC chromatogram for measuring the content of SN-38 of various structures in the product prepared in Example 1. [Figure 4] 1 is a typical SEC-HPLC chromatogram of the measurement of albumin aggregates in the product prepared in Example 1. [Figure 5] 1 shows XRD spectra of the freeze-dried product prepared in Example 2, SN-38, and HSA. [Figure 6] 1 shows the results of the decay experiment carried out under gradient dilution in Example 19. [Figure 7] 1 shows the results of the decay experiment carried out under gradient dilution in Example 20. [Figure 8] 2 shows the weight change in animals administered the rHA-SN-38 product of Example 1 in Example 21. [Figure 9] 1 shows the results of an in vivo experiment in which the product of Example 1 inhibited human triple-negative breast cancer MDA-MB-23. [Figure 10] 1 shows the results of the product of Example 2 inhibiting human colon cancer HT-29 in an in vivo experiment. [Figure 11] 1 shows the changes in body weight of test animals in Example 24. [Figure 12] 1 shows the change in tumor volume in test animals in Example 24. [Figure 13] 1 shows the changes in body weight of test animals in Example 25. [Figure 14] 1 shows the changes in tumor volume in test animals in Example 25. [Figure 15] 1 shows the changes in body weight of test animals in Example 26. [Figure 16] 1 shows the changes in tumor volume in test animals in Example 26. [Figure 17] 1 shows the changes in body weight of test animals in Example 27. [Figure 18] 1 shows the changes in tumor volume in test animals in Example 27. [Figure 19] 1 shows the changes in body weight of test animals in Example 28. [Figure 20] 1 shows the changes in tumor volume in test animals in Example 28. [Figure 21] 1 shows the results of a formulation containing Span 20 prepared in Example 30 in a disintegration experiment conducted under gradient dilution. [Figure 22] 1 shows the results of a formulation without Span 20 prepared in Example 30 in a disintegration experiment conducted under gradient dilution. [Figure 23] 1 shows the results of a formulation containing Span 20 prepared in Example 30 as observed by cryogenic transmission electron microscopy. [Figure 24] 1 shows the results of the Span 20-free formulation prepared in Example 30 as observed by cryogenic transmission electron microscopy. [Figure 25] 1 shows the changes in tumor volume in test animals in Example 34. [Figure 26] 1 shows the changes in body weight of test animals in Example 34. [Figure 27]1 shows the tumor mass diameters of test animals in Example 34. [Figure 28] 1 shows the changes in body weight of test animals in Example 38. [Figure 29] 1 shows the changes in tumor volume in test animals in Example 38. [Figure 30] 1 shows the changes in tumor volume in test animals in Example 38. [Figure 31] 1 shows the changes in body weight of test animals in Example 39. [Figure 32] 1 shows the changes in tumor volume in test animals in Example 39. [Figure 33] 1 shows the changes in tumor volume in test animals in Example 39. Detailed Description of the Invention

[0016] definition Unless otherwise defined below, all technical and scientific terms used herein are intended to have the same meaning as commonly understood by those skilled in the art. References to technology used herein are intended to refer to technology as commonly understood in the art (including variations of the technology, or substitutions by equivalent technology that would be apparent to those skilled in the art). Although the following terms will be readily understood by those skilled in the art, the following definitions are nevertheless provided to better illustrate the present invention.

[0017] The term "nanoparticle" refers to a particle having a nanoscale size in at least one dimension (e.g., one, two, or three dimensions), e.g., a size of about 1 nm, about 10 nm, about 100 nm, or about 200-300 nm, preferably 200 nm or less.

[0018] The term "nanocrystals" refers to crystals having a particle size of 1 to 1000 nm, in particular 50 to 300 nm, which may be monocrystalline or polycrystalline.

[0019] The term "vesicle" refers to an organized molecular assembly having an outer layer structure that can spontaneously form when dispersed in an aqueous phase.

[0020] The term "Span 20," also known as sorbitan monolaurate or Span 20, is a surfactant.

[0021] The term "aqueous composition" refers to a water-based composition in liquid or semi-solid form, preferably in liquid form, including but not limited to solutions (e.g., solutions of protein nanoparticles), colloids, emulsions, and suspensions.

[0022] The terms "drug loading (LD)" and "encapsulation efficiency (EE)" can be calculated according to the formulas shown in the Examples.

[0023] The term "organic acid" includes saturated or unsaturated fatty acids having 1 to 24 carbon atoms, particularly short-chain fatty acids having 2 to 4 carbon atoms, medium-chain fatty acids having 6 to 12 carbon atoms, and long-chain fatty acids having 14 to 24 carbon atoms; as well as aromatic carboxylic acids. Particular mention may be made of octanoic acid. "Long-chain fatty acids" include, but are not limited to, palmitic acid (C16:0), stearic acid (C18:0), oleic acid (C18:1), linoleic acid (C18:2), α-linolenic acid (C18:3), arachidonic acid (C20:4), thymnodonic acid (C20:5), and docosahexaenoic acid (C22:6). Particular mention may be made of palmitic acid and stearic acid.

[0024] As used herein, the terms "include," "comprise," "having," "contain," or "relating to," and other variations thereof, are inclusive or open-ended and do not exclude additional, unrecited elements or method steps, although these additional, unrecited elements or method steps are not necessarily present (i.e., these terms also encompass the terms "consisting essentially of" and "consisting of").

[0025] The term "about" means within ±10%, preferably within ±5%, and more preferably within ±2% of the stated value.

[0026] Numerical ranges recited herein are intended to include all subranges encompassed therein. For example, a range of "1 to 10" should be interpreted to include not only the explicitly stated values of 1 to 10, but also any single value within the range of 1 to 10 (e.g., 2, 3, 4, 5, 6, 7, 8, and 9) and any subrange (e.g., 1 to 2, 1.5 to 2.5, 1 to 3, 1.5 to 3.5, 2.5 to 4, and 3 to 4.5). This principle also applies to ranges using only one value as the minimum or maximum value.

[0027] All documents mentioned throughout this specification are incorporated herein by reference in their entirety.

[0028] In a first aspect, the present application provides a composition comprising SN-38, a lipid, albumin, and Span 20, wherein the composition comprises nanoparticles, wherein the albumin encapsulates at least a portion of the SN-38 and optionally at least a portion of the lipid; lipid:SN-38 is about (0.1-10):1 (w:w); albumin:SN-38 is about (1-100):1 (w:w); Span 20:SN-38 is about (3-60):100 (w:w); The composition is characterized in that the lipid is selected from cholesterol, cholesterol derivatives, cholesterol analogs, and fatty acid esters, and any combination of two or more thereof.

[0029] In some embodiments, the lipid:SN-38 ratio is about (0.5-6):1 (w:w), e.g., about (0.5-5):1 (w:w), about (0.5-3):1 (w:w), about (1-4):1 (w:w), about (1.2-4):1 (w:w), about (1.4-2):1 (w:w), about (1.5-2.5):1 (w:w), or about 1:1 (w:w).

[0030] In some embodiments, the ratio of albumin:SN-38 is about (1-50):1 (w:w), e.g., about (3-25):1 (w:w), about (5-25):1 (w:w), about (5-20):1 (w:w), or about (5-18):1 (w:w), about (6-15):1 (w:w), about (7-15):1 (w:w), about (6-12):1 (w:w), about (7-12):1 (w:w), about (9-11):1 (w:w), or about 10:1 (w:w).

[0031] In some embodiments, the Span 20:SN-38 ratio is about (4-60):100(w:w), e.g., about (5-60):100(w:w), about (6-60):100(w:w), about (7-55):100(w:w), about (8-50):100(w:w), about (10-45):100(w:w), about (12-40):100(w:w), about (14-35):100(w:w), about (15-30):100(w:w), about (16-25):100(w:w), or about (18-20):100(w:w).

[0032] In some embodiments, the albumin:lipid ratio is about (1-100):1 (w:w), such as about (2-20):1 (w:w), about (3-15):1 (w:w), or about (5-10):1 (w:w), such as about 7:1 (w:w) or about 10:1 (w:w).

[0033] In some embodiments, the SN-38 content is about 1% to about 25% w / w of the total amount of SN-38, lipid, and albumin in the composition. In some embodiments, the lipid content is about 1% to about 35% w / w of the total amount of SN-38, lipid, and albumin in the composition. In some embodiments, the albumin content is about 50% to about 98% w / w of the total amount of SN-38, lipid, and albumin in the composition.

[0034] In some embodiments, the SN-38 content is about 3% to about 20% by weight of the total amount of SN-38, lipid, and albumin in the composition. In some embodiments, the lipid content is about 2% to about 30% by weight of the total amount of SN-38, lipid, and albumin in the composition. In some embodiments, the albumin content is about 55% to about 95% by weight of the total amount of SN-38, lipid, and albumin in the composition.

[0035] In some embodiments, the SN-38 content is about 3 w / w% to about 15 w / w%, e.g., about 4 w / w%, about 5 w / w%, about 6 w / w%, about 6.5 w / w%, about 7 w / w%, about 7.5 w / w%, about 8 w / w%, about 8.5 w / w%, about 9 w / w%, about 10 w / w%, about 11 w / w%, about 12 w / w%, about 13 w / w%, or about 14 w / w% of the total amount of SN-38, lipid, and albumin in the composition.

[0036] In some embodiments, the lipid content is about 3 w / w% to about 30 w / w%, e.g., about 4 w / w%, about 5 w / w%, about 6 w / w%, about 7 w / w%, about 8 w / w%, about 8.5 w / w%, about 9 w / w%, about 9.5 w / w%, about 10 w / w%, about 10.5 w / w%, or about 11 w / w%, about 11.5 w / w%, about 12 w / w%, about 12.5 w / w%, about 13 w / w%, about 13.5 w / w%, about 14 w / w%, about 15 w / w%, about 16 w / w%, about 17 w / w%, about 18 w / w%, about 19 w / w%, about 20 w / w%, about 21 w / w%, about 24 w / w%, about 26 w / w%, or about 28 w / w%.

[0037] In some embodiments, the albumin content is about 60 w / w% to about 94 w / w%, e.g., about 64 w / w% to about 93 w / w%, about 66 w / w% to about 92 w / w%, about 68 w / w% to about 91 w / w%, about 70 w / w% to about 90 w / w%, about 75 w / w% to about 90 w / w%, about 75 w / w%, about 76 w / w%, about 77 w / w%, about 78 w / w%, about 79 w / w%, about 80 w / w%, about 81 w / w%, about 82 w / w%, about 83 w / w%, about 84 w / w%, about 85 w / w%, about 86 w / w%, about 87 w / w%, about 88 w / w%, or about 89 w / w% of the total amount of SN-38, lipid, and albumin in the composition.

[0038] Without wishing to be bound by theory, the inventors have discovered that increasing the amount of lipid used tends to result in a composition that exhibits a decreased mean particle size, increased encapsulation efficiency, and increased availability of SN-38; increasing the amount of albumin used may decrease drug loading; and excessively high albumin content may increase nanoparticle size, affecting druggability. In one embodiment, the lipid content is about 5 wt% to about 24 wt% of the total amount of SN-38, lipid, and albumin in the composition. In one embodiment, the albumin content is about 60 wt% to about 90 wt% of the total amount of SN-38, lipid, and albumin in the composition.

[0039] On the other hand, excessively high content of lipid components in the composition may cause adverse effects to individuals.From the viewpoint of reducing lipid intake caused by administration of the composition, it is expected that the lipid content in the composition is preferably about 18 w / w% or less, for example, about 16 w / w% or less or about 14 w / w% or less, based on the total amount of SN-38, lipid, and albumin in the composition.However, this does not mean that compositions with higher lipid content are undesirable.

[0040] In some embodiments, the content of Span 20 relative to the total amount of SN-38, lipid, albumin, and Span 20 in the composition is about 0.03 w / w% to about 12 w / w%, e.g., about 0.06 w / w% to about 10 w / w%, 0.08 w / w% to about 9 w / w%, about 0.1 w / w% to about 8 w / w%, about 0.2 w / w% to about 7 w / w%, about 0.4 w / w% to about 6 w / w%, about 0.6 w / w% to about 5 w / w%, about 0.8 w / w% to about 4 w / w%, or about 1 w / w% to about 2 w / w%.

[0041] In some embodiments, the lipid:SN-38 ratio is about (1-10):1 (w:w), e.g., about (1-8):1 (w:w), about (1-6):1 (w:w), about (1-5):1 (w:w), about (1-4.5):1 (w:w), about (1-4):1 (w:w), about (1.2-3.8):1 (w:w), about 1:1 (w:w), about (1.4-3.6):1 (w:w), about (1.6-3.4):1 (w:w), about (1.8-3.2):1 (w:w), about (2-3):1 (w:w), about (2.2-2.8):1 (w:w), about (2.4-2.6):1 (w:w), about 1:1 (w:w), or about 2.5:1 (w:w). In some other embodiments, the lipid:SN-38 ratio is about (0.8-1.8):1 (w:w), about (0.9-1.7):1 (w:w), or about (1-1.4):1.

[0042] In some embodiments, the albumin:SN-38 ratio is about (5-40):1 (w:w), e.g., about (less than 5-40):1 (w:w), about (5-35):1 (w:w), about (5-30):1 (w:w), about (8-25):1 (w:w), about (10-22.5):1 (w:w), about (12.5-20):1 (w:w), about (15-17.5):1 (w:w), about (16-18):1 (w:w), or about 10:1 (w:w). In some other embodiments, the albumin:SN-38 ratio is about (9-21):1 (w:w), about (9-20):1 (w:w), about (11-18):1 (w:w), or about (11.1-17.3):1 (w:w).

[0043] In some embodiments, the Span 20:SN-38 ratio is about (5-60):100(w:w), e.g., about (6-60):100(w:w), about (7-55):100(w:w), about (8-50):100(w:w), about (10-45):100(w:w), about (12-40):100(w:w), about (14-35):100(w:w), about (15-30):100(w:w), about (16-25):100(w:w), or about (18-20):100(w:w). In some other embodiments, the Span 20:SN-38 ratio is about (5-10):100 (w:w), about (5-9):100 (w:w), about (6-8.6):100 (w:w), about (6-8):100 (w:w), or about (6.5-7):100 (w:w).

[0044] In some embodiments, the albumin:lipid ratio is about (1-40):1 (w:w), e.g., about (less than 1-40):1 (w:w), about (2-35):1 (w:w), about (3-15):1 (w:w), about (5-10):1 (w:w), or about (6-8):1 (w:w), e.g., about 7:1 (w:w) or about 10:1 (w:w). In some embodiments, the albumin:lipid ratio is about (6-21):1 (w:w), about (6.7-13):1 (w:w), about (7-13):1 (w:w), or about (11-12.7):1 (w:w).

[0045] In some embodiments, the SN-38 content is about 2 wt% to about 16 wt% of the total amount of SN-38, lipid, and albumin in the composition. In some embodiments, the lipid content is about 2 wt% to about 35 wt% of the total amount of SN-38, lipid, and albumin in the composition. In some embodiments, the albumin content is about 75 wt% to about 96 wt% of the total amount of SN-38, lipid, and albumin in the composition.

[0046] In some embodiments, the SN-38 content is about 2.5 w / w% to about 15 w / w%, e.g., about 4 w / w% to about 10 w / w%, about 4.5 w / w% to about 9.5 w / w%, about 5 w / w% to about 9 w / w%, or about 7.5 w / w% to about 8 w / w%, relative to the total amount of SN-38, lipid, and albumin in the composition. In some embodiments, the lipid content in the composition is about 2.5 w / w% to about 30 w / w%, e.g., about 4 w / w% to about 12.5 w / w%, about 4.5 w / w% to about 12 w / w%, about 7 w / w% to about 10 w / w%, or about 7.5 w / w% to about 8 w / w%, relative to the total amount of SN-38, lipid, and albumin in the composition. In some embodiments, the albumin content of the composition is about 76 w / w% to about 95 w / w%, e.g., about 78 w / w% to about 93 w / w%, about 79 w / w% to about 91.5 w / w%, about 80 w / w% to about 90 w / w%, about 82 w / w% to about 89 w / w%, about 84 w / w% to about 88 w / w%, or about 84.5 w / w% to about 87.5 w / w%, relative to the total amount of SN-38, lipid, and albumin in the composition.

[0047] In some embodiments, the SN-38 content is about 3 w / w% to about 14 w / w%, e.g., about 3.5 w / w% to about 12 w / w%, about 4 w / w%, about 4.2 w / w%, about 4.5 w / w%, about 4.6 w / w%, about 4.8 w / w%, about 5 w / w%, about 5.5 w / w%, about 6 w / w%, or about 7 w / w% of the total amount of SN-38, lipid, and albumin in the composition. w / w%, about 6.5 w / w%, about 7 w / w%, about 7.5 w / w%, about 7.6 w / w%, about 7.8 w / w%, about 8 w / w%, about 8.5 w / w%, about 9 w / w%, about 9.2 w / w%, about 9.5 w / w%, about 9.6 w / w%, about 9.8 w / w%, about 10 w / w%, about 10.5 w / w%, about 11 w / w%, or about 11.5 w / w%.

[0048] In some embodiments, the lipid content is about 4 w / w% to about 25 w / w%, e.g., about 4.5 w / w% to about 20 w / w%, about 4.3 w / w%, about 4.5 w / w%, about 4.7 w / w%, about 5 w / w%, about 5.5 w / w%, about 6 w / w%, about 6.5 w / w%, about 7 w / w%, about 7.5 w / w%, about 7.6 w / w%, about 7.8 w / w%, about 8 w / w%, 8.5 w / w%, about 9 w / w%, or about 9.5 w / w% of the total amount of SN-38, lipid, and albumin in the composition. , about 10 w / w%, about 10.5 w / w%, about 11 w / w%, about 11.5 w / w%, about 12 w / w%, about 12.1 w / w%, about 12.3 w / w%, about 12.5 w / w%, about 13 w / w%, about 13.5 w / w%, about 14 w / w%, about 14.5 w / w%, about 15 w / w%, about 15.5 w / w%, about 16 w / w%, about 16.5 w / w%, about 17 w / w%, about 17.5 w / w%, about 18 w / w%, about 18.5 w / w%, about 19 w / w%, or about 19.5 w / w%.

[0049] In some embodiments, the albumin content of the total amount of SN-38, lipid, and albumin in the composition is about 78 w / w% to about 92 w / w%, for example, about 79 w / w%, about 79.2 w / w%, about 79.4 w / w%, about 79.6 w / w%, about 79.8 w / w%, about 80 w / w%, about 81 w / w%, about 82 w / w%, about 83 w / w%, about 84 w / w%, or about 84.3 w / w%, about 84.5 w / w%, about 84.7 w / w%, about 84.9 w / w%, about 85 w / w%, about 86 w / w%, about 87 w / w%, about 87.3 w / w%, about 87.5 w / w%, about 87.7 w / w%, about 87.9 w / w%, about 88 w / w%, about 89 w / w%, about 90 w / w%, about 91 w / w%, about 91.3 w / w%, or about 91.5 w / w%.

[0050] In some embodiments, the content of Span 20 relative to the total amount of SN-38, lipid, albumin, and Span 20 in the composition is about 0.14 w / w% to about 5 w / w%, e.g., about 0.2 w / w% to about 2.5 w / w%, about 0.22 w / w% to about 2.0 w / w%, about 0.24 w / w% to about 2 w / w%, about 0.26 w / w% to about 1.5 w / w%, about 0.28 w / w% to about 1.0 w / w%, about 0.3 w / w% to about 0.9 w / w%, or about 0.32 w / w% to about 0.8 w / w%. w%, about 0.34 w / w% to about 0.7 w / w%, about 0.36 w / w% to about 0.6 w / w%, about 0.38 w / w% to about 0.58 w / w%, about 0.4 w / w% to about 0.56 w / w%, about 0.42 w / w% to about 0.54 w / w%, about 0.44 w / w% to about 0.52 w / w%, about 0.46 w / w%, about 0.48 w / w%, or about 0.5 w / w%. In some other embodiments, the Span 20 content is about 0.2 w / w% to about 0.8 w / w%, about 0.24 w / w% to about 0.7 w / w%, about 0.26 w / w% to about 0.7 w / w%, about 0.3 w / w% to about 0.65 w / w%, about 0.36 w / w% to about 0.6 w / w%, about 0.4 w / w% to about 0.58 w / w%, about 0.44 w / w% to about 0.56 w / w%, about 0.48 w / w% to about 0.54 w / w%, or about 0.5 w / w% to about 0.52 w / w%.

[0051] In some embodiments, the SN-38 present in the nanoparticles comprises at least about 1 wt% or at least about 2 wt% of the total amount of SN-38, lipid, and albumin in the composition, e.g., at least about 3 wt%, about 3 wt% to about 13 wt%, about 4 wt% to about 12 wt%, about 4 wt%, about 5 wt%, about 6 wt%, about 7 wt%, about 8 wt%, about 9 wt%, about 10 wt%, or about 11 wt%.

[0052] In some embodiments, the SN-38 present in the nanoparticles comprises about 80 w / w% to about 99 w / w% of the total amount of SN-38 in the composition, e.g., about 88 w / w% to about 98 w / w%, about 89 w / w%, about 90 w / w%, about 91 w / w%, about 92 w / w%, about 93 w / w%, about 94 w / w%, about 95 w / w%, about 96 w / w%, or about 97 w / w%.

[0053] In some embodiments, the cholesterol derivative is selected from esters formed by cholesterol and organic acids, preferably selected from cholesteryl palmitate, cholesteryl caprylate, and combinations thereof.

[0054] In some embodiments, the cholesterol analog is selected from vitamin D2, vitamin D3, and combinations thereof.

[0055] In some embodiments, the fatty acid ester is selected from fatty acid glycerides, preferably long chain fatty acid glycerides, preferably glyceryl stearate, more preferably glyceryl monostearate.

[0056] In some preferred embodiments, the lipid is selected from cholesterol, cholesteryl palmitate, cholesteryl caprylate, vitamin D2, vitamin D3, glyceryl monostearate, and any combination of two or more thereof.

[0057] In some preferred embodiments, the lipid is selected from cholesterol, cholesteryl palmitate, vitamin D3, glyceryl monostearate, and any combination of two or more thereof.

[0058] In some preferred embodiments, the lipid is cholesterol, cholesteryl palmitate, vitamin D3, or glyceryl monostearate; a mixture of cholesterol and cholesteryl palmitate; a mixture of cholesterol and vitamin D3; a mixture of cholesterol and glyceryl monostearate; or a mixture of cholesteryl palmitate and glyceryl monostearate.

[0059] In some more preferred embodiments, the lipid is cholesterol. Preferably, cholesterol is used as the only lipid. In some of these embodiments, the cholesterol:SN-38 ratio is about (1-6):1 (w:w), e.g., about (1.2-5):1 (w:w), e.g., about (1.4-4):1 (w:w), about 3:1 (w:w), about 2:1 (w:w), or about 1:1 (w:w). In some other embodiments, the cholesterol:SN-38 ratio is about (0.8-1.8):1 (w:w), about (0.9-1.7):1 (w:w), or about (1-1.4):1. In some of these embodiments, the albumin:SN-38 ratio is about (3-25):1 (w:w), e.g., about (4-20):1 (w:w), about (5-15):1 (w:w), about (6-12):1 (w:w), about (7-12):1 (w:w), about (9-11):1 (w:w), or about 10:1 (w:w). In some other embodiments, the albumin:SN-38 ratio is about (9-21):1 (w:w), about (9-20):1 (w:w), about (11-18):1 (w:w), or about (11.1-17.3):1 (w:w). In some of these embodiments, the albumin:cholesterol ratio is about (2-20):1 (w:w), e.g., about (3-15):1 (w:w), about (5-10):1 (w:w), or about 7:1 (w:w). In some other embodiments, the albumin:cholesterol ratio is about (6-21):1 (w:w), about (6.7-13):1 (w:w), about (7-13):1 (w:w), or about (11-12.7):1 (w:w).

[0060] In some of these embodiments, the SN-38 content is about 3 w / w% to about 15 w / w%, e.g., about 4 w / w% to about 15 w / w%, about 6 w / w% to about 10 w / w%, or about 8 w / w% to about 12 w / w%, relative to the total amount of SN-38, cholesterol, and albumin in the composition. In some other embodiments, the SN-38 content is about 4 w / w% to about 10 w / w%, about 4.5 w / w% to about 9.5 w / w%, about 5 w / w% to about 9 w / w%, or about 7.5 w / w% to about 8 w / w%. In some of these embodiments, the cholesterol content is about 5 w / w% to about 25 w / w%, e.g., about 6 w / w% to about 22 w / w%, or about 15 w / w% to about 20 w / w%, relative to the total amount of SN-38, cholesterol, and albumin in the composition. In some other embodiments, the cholesterol content is about 4% to about 12.5%, about 4.5% to about 12%, about 7% to about 10%, or about 7.5% to about 8% by weight. In some of these embodiments, the albumin content is about 64% to about 90%, e.g., about 70% to about 90% by weight, based on the total amount of SN-38, cholesterol, and albumin in the composition. In some other embodiments, the albumin content is about 78% to about 93%, about 79% to about 91.5%, about 80% to about 90%, about 82% to about 89%, about 84% to about 88%, or about 84.5% to about 87.5% by weight. In some of these embodiments, the SN-38 present in the nanoparticles comprises at least about 3% by weight, e.g., about 3% to about 13%, about 4% to about 12%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, or about 11% by weight of the total amount of SN-38, cholesterol, and albumin in the composition.

[0061] In some other embodiments, the lipid is cholesteryl palmitate. In some embodiments, the lipid is vitamin D3. In some embodiments, the lipid is glyceryl monostearate. In some embodiments, cholesteryl palmitate, vitamin D3, or glyceryl monostearate are used as the only lipid.

[0062] It may also be implied that the lipid is a mixture of cholesterol and glyceryl monostearate, for example, about (0.2-5):1 (w:w), about (0.5-3):1 (w:w), about (0.5-2):1 (w:w), or about 1:1 (w:w).

[0063] In some embodiments, the lipid is a mixture of cholesteryl palmitate and glyceryl monostearate, e.g., about (0.2-5):1 (w:w), about (0.5-3):1 (w:w), about (0.5-2):1 (w:w), or about 1:1 (w:w).

[0064] In some embodiments, the lipid is a mixture of cholesterol and cholesteryl palmitate, e.g., cholesterol:cholesteryl palmitate in a ratio of about (0.2-5):1 (w:w), about (0.5-3):1 (w:w), about (0.5-2):1 (w:w), or about 1:1 (w:w).

[0065] In embodiments including lipids other than cholesterol, or a combination of cholesterol and another lipid, the lipid:SN-38 ratio is, for example, (1-6):1 (w:w), about (1.2-5):1 (w:w), about (1.4-4.5):1 (w:w), about 4.3:1 (w:w), about 3:1 (w:w), about 1:1 (w:w), or about 2:1 (w:w). In some of these embodiments, the albumin:SN-38 ratio is, for example, about (5-25):1 (w:w), about (10-20):1 (w:w), about (6-15):1 (w:w), about (7-15):1 (w:w), about (9-12):1 (w:w), about (9-11):1 (w:w), about 9.5:1 (w:w), or about 10:1 (w:w). In some of these embodiments, the albumin:lipid ratio is, for example, about (2-10):1 (w:w), about (3-7):1 (w:w), or about (4-6):1 (w:w). In some of these embodiments, the SN-38 content is, for example, about 5 w / w% to about 15 w / w%, about 6 w / w% to about 12 w / w%, about 7 w / w% to about 10 w / w%, about 8 w / w%, or about 9 w / w% relative to the total amount of SN-38, lipid, and albumin in the composition. In some of these embodiments, the lipid content is, for example, about 5% to about 32%, about 10% to about 30%, about 18%, about 20%, about 22%, about 24%, about 26%, about 28%, or about 29% by weight of the total amount of SN-38, lipid, and albumin in the composition. In some of these embodiments, the albumin content is, for example, about 60% to about 90%, about 64% to about 85%, about 70% to about 80%, or about 75% by weight of the total amount of SN-38, lipid, and albumin in the composition. In some of these embodiments, the SN-38 present in the nanoparticles comprises at least about 3 wt% of the total amount of SN-38, lipid, and albumin in the composition, e.g., about 3 wt% to about 10 wt%, about 4 wt% to about 9 wt%, about 4 wt%, about 5 wt%, about 6 wt%, about 7 wt%, or about 8 wt%.

[0066] In some of the above embodiments, the composition is in liquid, semi-solid, or solid form.

[0067] In some embodiments, the composition is in solid form, preferably in powder form. More preferably, the composition is a lyophilized powder.

[0068] In some of these embodiments, the SN-38 is present in the composition in amorphous and / or nanocrystalline form, preferably as determined by electron microscopy or X-ray diffraction (Cu-Kα) analysis. The nanocrystals may have a particle size ranging from about 30 to 500 nm, preferably about 50 to 200 nm. The nanocrystalline form of SN-38 comprises 75% or more of the total SN-38, e.g., 80% or more, 85% or more, or 90% or more.

[0069] Without wishing to be bound by theory, in the compositions of the present application, Span 20 plays a role in improving the stability of the composition, adjusting the particle size of the nanoparticles, and increasing the dispersibility of the nanoparticles.

[0070] In some embodiments, the composition does not contain an additional stabilizer. In some other embodiments, the composition preferably further comprises an additional stabilizer, such as a lyophilization stabilizer, present in an amount such that when the composition is reconstituted to form an aqueous composition (including solutions and emulsions), the content of the additional stabilizer is at least about 2 w / v%, e.g., at least about 3 w / v%, e.g., at least about 5 w / v%, about 5 w / v% to about 30 w / v%, about 10 w / v% to about 25 w / v%, or about 15 w / v% to about 20 w / v%.

[0071] In some of these embodiments, the composition is present in an amount of about 60 w / w% to about 98 w / w%, e.g., about 65 w / w% to about 97 w / w%, about 68 w / w% to about 96 w / w%, about 69 w / w% to about 95 w / w%, about 70 w / w% to about 94 w / w%, about 71 w / w% to about 93 w / w%, about 72 w / w% to about 92 w / w%, about 73 w / w%, or about 74 w / w%, about 75 w / w%, about 76 w / w%, about 77 w / w%, about 78 w / w%, about 79 w / w%, about 80 w / w%, about 81 w / w%, about 82 w / w%, about 83 w / w%, about 84 w / w%, about 85 w / w%, about 86 w / w%, about 87 w / w%, about 88 w / w%, about 89 w / w%, about 90 w / w%, or about 91 w / w% of an additional stabilizer.

[0072] In some embodiments, when the composition is reconstituted to form an aqueous composition (including solutions and emulsions) having an SN-38 content of about 0.1 μg / mL to about 30.0 mg / mL (or the SN-38 content values set forth below for aqueous compositions), the nanoparticles have an average particle size of about 50-200 nm, e.g., about 90-150 nm, about 95-140 nm, about 100-130 nm, about 105-125 nm, or about 110-120 nm.

[0073] In some other embodiments, the composition is an aqueous composition in liquid form, including solutions and emulsions.

[0074] In some embodiments, the liquid form of the composition comprises SN-38 in the form of nanocrystals and / or vesicles. The nanocrystals may have a particle size range of about 30-500 nm, preferably about 50-200 nm.

[0075] In some of these embodiments, the composition is in the form of a solution; in some of these embodiments, the composition is in the form of an emulsion.

[0076] The present inventors have discovered that the aqueous compositions of the present application exhibit excellent dilution stability. In some embodiments, the nanoparticles do not disintegrate when the composition is diluted (e.g., using 1xPBS at a pH of about 7.4) to produce an SN-38 content of about 4 μg / mL or less, e.g., about 2 μg / mL or less, about 1 μg / mL or less, or about 0.4 μg / mL or less, e.g., about 0.1 μg / mL or 0.04 μg / mL, in the diluted composition. Due to their excellent stability, the aqueous compositions can exist as concentrated or diluted solutions.

[0077] Thus, the various components of the aqueous composition may have a wide range of content. In some of these embodiments, the SN-38 content, relative to the total volume of the composition, is about 0.1 μg / mL to about 30.0 mg / mL, about 0.2 μg / mL to about 27.0 mg / mL, about 0.5 μg / mL to about 24.0 mg / mL, about 1.0 μg / mL to about 21.0 mg / mL, about 5.0 μg / mL to about 18.0 mg / mL, about 10.0 μg / mL to about 15.0 mg / mL, about 20.0 μg / mL to about 12 mg / mL, about 25.0 μg / mL to about 9 mg / mL, about 50.0 μg / mL to about 6.0 mg / mL, or about 100.0 μg / mL to about 3.0 mg / mL.

[0078] In some embodiments, the lipid content, relative to the total volume of the composition, is about 0.05 μg / mL to about 100.0 mg / mL, about 0.1 μg / mL to about 90.0 mg / mL, about 0.25 μg / mL to about 80.0 mg / mL, about 0.5 μg / mL to about 70.0 mg / mL, about 2.5 μg / mL to about 60.0 mg / mL, about 5.0 μg / mL to about 50.0 mg / mL, about 10.0 μg / mL to about 40.0 mg / mL, about 12.5 μg / mL to about 30.0 mg / mL, about 25.0 μg / mL to about 20.0 mg / mL, or about 50.0 μg / mL to about 10.0 mg / mL.

[0079] In some embodiments, the albumin content, relative to the total volume of the composition, is about 3.0 μg / mL to about 300.0 mg / mL, about 6.0 μg / mL to about 270.0 mg / mL, about 15.0 μg / mL to about 240.0 mg / mL, about 30.0 μg / mL to about 210.0 mg / mL, about 150.0 μg / mL to about 180.0 mg / mL, about 300.0 μg / mL to about 150.0 mg / mL, about 600.0 μg / mL to about 120.0 mg / mL, about 750.0 μg / mL to about 90.0 mg / mL, about 1500.0 μg / mL to about 60.0 mg / mL, or about 3.0 mg / mL to about 30.0 mg / mL.

[0080] Generally, the content of SN-38 relative to the total volume of the composition may be about 100.0 μg / mL to about 3.0 mg / mL, for example, about 200.0 μg / mL to about 2.5 mg / mL, about 300.0 μg / mL to about 2.0 mg / mL, about 400.0 μg / mL to about 1.5 mg / mL, about 500.0 μg / mL to about 1.0 mg / mL, or about 600 μg / mL to about 800 μg / mL; and / or the lipid content may be from about 50.0 μg / mL to about 10.0 mg / mL, e.g., from about 100.0 μg / mL to about 8.0 mg / mL, from about 200.0 μg / mL to about 6.0 mg / mL, from about 300.0 μg / mL to about 4.0 mg / mL, from about 400.0 μg / mL to about 3.0 mg / mL, from about 500.0 μg / mL to about 2.5 mg / mL, from about 600.0 μg / mL to about 2.0 mg / mL, from about 700.0 μg / mL to about 1.5 mg / mL, from about 800 μg / mL to about 1.0 mg / mL, or from about 200 μg / mL to about 1.5 mg / mL; and / or The albumin content can be about 3.0 mg / mL to about 30.0 mg / mL, for example, about 4.0 mg / mL to about 25.0 mg / mL, about 5.0 mg / mL to about 20.0 mg / mL, about 6.0 mg / mL to about 15.0 mg / mL, about 7.0 mg / mL to about 12.0 mg / mL, or about 8.0 mg / mL to about 10.0 mg / mL.

[0081] Ideally, in aqueous compositions (including solutions and emulsions), after a period of storage prior to administration, the nanoparticles have an average particle size of about 200 nm or less, e.g., about 150 nm or less, and preferably the average particle size of the nanoparticles still meets the above requirements. The inventors have discovered that the aqueous compositions of the present application exhibit these excellent properties.

[0082] In some embodiments, the nanoparticles have an average particle size of about 50-200 nm, such as about 90-150 nm or about 100-130 nm.

[0083] In some embodiments, after storage at 4° C. for 24 hours, the nanoparticles have an average particle size of about 50-200 nm, eg, about 90-150 nm or about 100-130 nm.

[0084] In some embodiments, the nanoparticles have a particle size distribution index (PDI) of about 0.30 or less, eg, about 0.2 or less, about 0.10 or less, or about 0.01 or less.

[0085] In some embodiments, the composition has a zeta potential of about −35 mV to about −20 mV, for example, about −31 mV.

[0086] In some of these embodiments, the composition does not contain an additional stabilizer. In some of these embodiments, the composition further contains an additional stabilizer in an amount of at least about 2 w / v%, preferably at least about 3 w / v%, e.g., at least about 5 w / v%, about 5 w / v% to about 30 w / v%, about 10 w / v% to about 25 w / v%, or about 15 w / v% to about 20 w / v%, based on the total amount of the composition.

[0087] The additional stabilizer may be selected from albumin (e.g., human serum albumin, recombinant human albumin, bovine serum albumin, and skim milk powder), monosaccharides, disaccharides, polysaccharides, and any combination thereof, and preferably from glucose and sucrose, preferably sucrose.

[0088] The use of an additional stabilizer helps maintain the average particle size of the nanoparticles. The inventors have discovered that the presence of an additional stabilizer reduces the degree of increase in the average particle size of nanoparticles in an aqueous composition after 24 hours of storage at 4°C compared to when the additional stabilizer is absent. At the same time, for compositions ultimately obtained in the form of lyophilized powder, the use of an additional stabilizer, particularly a saccharide stabilizer, provides additional advantages. This is because the additional stabilizer can simultaneously act as a lyophilization excipient during the lyophilization of the aqueous composition, thereby avoiding the use of other lyophilization excipients used in the prior art, particularly albumin (e.g., HSA), which is cost-effective and helps reduce the risk of drug anaphylaxis. Therefore, in some embodiments, the composition preferably does not contain an additional lyophilization stabilizer. Of course, the composition may contain additional lyophilization excipients, such as one or more of sucrose, mannitol, lactose, maltose, trehalose, and dextran, if necessary.

[0089] In a first subset of the first aspect, the present application provides a composition comprising SN-38, a lipid, albumin, and Span 20, wherein the composition comprises nanoparticles, wherein the albumin encapsulates at least a portion of the SN-38 and optionally at least a portion of the lipid, and wherein the lipid is cholesterol; where: cholesterol:SN-38, in some embodiments, is about (1-3):1(w:w), e.g., about (1.2-2.5):1(w:w), about (1.4-2):1(w:w), about (1.5-2):1(w:w), about (1.3-1.8):1(w:w), about (1.4-1.6):1(w:w), about (1.5-1.7):1(w:w), about (1.2-1.5):1(w:w), about 1:1(w:w), or about (1.4-1.5):1(w:w), or in some other embodiments, about (0.8-1.8):1(w:w), about (0.9-1.7):1(w:w), or about (1-1.4):1; albumin:SN-38, in some embodiments, is about (5-15):1(w:w), e.g., about (5-12):1(w:w), about (6-12):1(w:w), or about (7-12):1(w:w), about (9-11):1(w:w), about (10-12):1(w:w), or about 11:1(w:w), or in some other embodiments, about (9-21):1(w:w), about (9-20):1(w:w), about (11-18):1(w:w), or about (11.1-17.3):1(w:w); albumin:cholesterol, in some embodiments, about (3-10):1(w:w), about (4-8):1(w:w), or about (5-7):1(w:w), or in some other embodiments, about (6-21):1(w:w), about (6.7-13):1(w:w), about (7-13):1(w:w), or about (11-12.7):1(w:w); A composition is provided, characterized in that Span 20:SN-38 is as described above.

[0090] In some embodiments, the SN-38 content is about 6% to about 14% by weight, e.g., about 6.5% to about 13%, about 7% to about 12%, about 7.5% to about 12%, about 8% to about 11%, about 8.5% to about 10%, or about 9% by weight, based on the total amount of SN-38, cholesterol, and albumin in the composition. In some other embodiments, the SN-38 content is about 4% to about 10%, about 4.5% to about 9.5%, about 5% to about 9%, or about 7.5% to about 8% by weight. In some embodiments, the cholesterol content is about 8 w / w% to about 18 w / w%, e.g., 8.5 w / w% to about 17 w / w%, about 9 w / w% to about 16 w / w%, about 9.5 w / w% to about 16 w / w%, about 10 w / w% to about 16 w / w%, about 10.5 w / w% to about 16 w / w%, about 11 w / w% to about 15 w / w%, about 11.5 w / w% to about 15 w / w%, about 12 w / w% to about 15 w / w%, about 12.5 w / w% to about 14 w / w%, or about 13 w / w% to about 13.5 w / w% relative to the total amount of SN-38, cholesterol, and albumin in the composition. In some other embodiments, the cholesterol content is about 4 w / w% to about 12.5 w / w%, about 4.5 w / w% to about 12 w / w%, about 7 w / w% to about 10 w / w%, or about 7.5 w / w% to about 8 w / w%. In some embodiments, the albumin content of the composition is about 66 w / w% to about 90 w / w%, e.g., about 68 w / w% to about 89 w / w, about 70 w / w% to about 88 w / w%, about 70 w / w% to about 87 w / w%, about 70 w / w% to about 86 w / w%, about 70 w / w% to about 85 w / w%, about 75 w / w% to about 85 w / w%, about 76 w / w%, about 77 w / w%, about 78 w / w%, about 79 w / w%, about 80 w / w%, about 81 w / w%, about 82 w / w%, about 83 w / w%, or about 84 w / w% of the total amount of SN-38, cholesterol, and albumin in the composition. In some other embodiments, the albumin content is about 78 w / w% to about 93 w / w%, about 79 w / w% to about 91.5 w / w%, about 80 w / w% to about 90 w / w%, about 82 w / w% to about 89 w / w%, about 84 w / w% to about 88 w / w%, or about 84.5 w / w% to about 87.5 w / w%.

[0091] In a second subset of the first aspect, the present application provides a composition comprising SN-38, a lipid, albumin, and Span 20, wherein the composition comprises nanoparticles, wherein the albumin encapsulates at least a portion of the SN-38 and optionally at least a portion of the lipid, and wherein the lipid is cholesterol; where: In some embodiments, cholesterol:SN-38 is about (1 to 5):1 (w:w), for example, about (1 to 4.5):1 (w:w), about (1 to 4):1 (w:w), about (1.2 to 3.8):1 (w:w), about (1.4 to 3.6):1 (w:w), about (1.6 to 3.4):1 (w:w), about (1.8 to 3.2):1 (w:w), about (2 to 3):1 (w:w), about (2.2-2.8):1 (w:w), about (2.4-2.6):1 (w:w), about 2.5:1 (w:w), or about 1:1 (w:w), or in some other embodiments about (0.8-1.8):1 (w:w), about (0.9-1.7):1 (w:w), or about (1-1.4):1; and / or and / or, the albumin:SN-38 ratio in some embodiments is about (5-25):1 (w:w), e.g., about (5-20):1 (w:w), about (6-19):1 (w:w), about (7-18):1 (w:w), about (8-16):1 (w:w), about (9-14):1 (w:w), or about (10-12):1 (w:w), or in some other embodiments, about (9-21):1 (w:w), about (9-20):1 (w:w), about (11-18):1 (w:w), or about (11.1-17.3):1 (w:w); and / or and / or, the albumin:cholesterol ratio in some embodiments is about (5-25):1(w:w), e.g., about (6-20):1(w:w), about (7-18):1(w:w), about (8-16):1(w:w), about (9-14):1(w:w), or about (10-12):1(w:w), or in some other embodiments, about (6-21):1(w:w), about (6.7-13):1(w:w), about (7-13):1(w:w), or about (11-12.7):1(w:w); and / or Compositions are provided in which the Span 20:SN-38 ratio is, in some embodiments, about (5-15):100 (w:w), e.g., about (6-12):100 (w:w), about (7-10):100 (w:w), or about 7.5:100 (w:w), or in some other embodiments, about (5-10):100 (w:w), about (5-9):100 (w:w), about (6-8.6):100 (w:w), about (6-8):100 (w:w), or about (6.5-7):100 (w:w).

[0092] In some of these embodiments, the SN-38 content is about 3 w / w% to about 10 w / w%, e.g., about 3.5 w / w% to about 9.5 w / w%, about 4 w / w%, about 4.5 w / w%, about 5 w / w%, about 5.5 w / w%, about 6 w / w%, about 6.5 w / w%, about 7 w / w%, about 7.5 w / w%, about 8 w / w%, about 8.5 w / w%, or about 9 w / w% of the total amount of SN-38, cholesterol, and albumin in the composition. In some other embodiments, the SN-38 content is about 4 w / w% to about 10 w / w%, about 4.5 w / w% to about 9.5 w / w%, about 5 w / w% to about 9 w / w%, or about 7.5 w / w% to about 8 w / w%.

[0093] In some of these embodiments, the cholesterol content is about 4 w / w% to about 18 w / w%, e.g., about 4.5 w / w% to about 17.5 w / w%, about 5 w / w%, about 5.5 w / w%, about 6 w / w%, about 6.5 w / w%, about 7 w / w%, about 7.5 w / w%, about 8 w / w%, 8.5 w / w%, or the like, relative to the total amount of SN-38, cholesterol, and albumin in the composition. In some other embodiments, the cholesterol content is about 4 w / w% to about 12.5 w / w%, about 4.5 w / w% to about 12 w / w%, about 7 w / w% to about 10 w / w%, or about 7.5 w / w% to about 8 w / w%.

[0094] In some of these embodiments, the albumin content is about 78 w / w% to about 92 w / w%, e.g., about 79 w / w%, about 80 w / w%, about 81 w / w%, about 82 w / w%, about 83 w / w%, about 84 w / w%, about 85 w / w%, about 86 w / w%, about 87 w / w%, about 88 w / w%, about 89 w / w%, about 90 w / w%, or about 91 w / w%, relative to the total amount of SN-38, cholesterol, and albumin in the composition. In some other embodiments, the albumin content is about 78 w / w% to about 93 w / w%, about 79 w / w% to about 91.5 w / w%, about 80 w / w% to about 90 w / w%, about 82 w / w% to about 89 w / w%, about 84 w / w% to about 88 w / w%, or about 84.5 w / w% to about 87.5 w / w%.

[0095] In some further embodiments, the present invention provides cholesterol:SN-38 is about (1-2.5):1 (w:w), for example about 1.25:1 (w:w) or about 1:1 (w:w); and / or albumin:SN-38 is about (8-25):1 (w:w), e.g., about (10-20):1 (w:w), about (12-18):1 (w:w), about (12.5-17):1 (w:w), about (15-16):1 (w:w), about 10:1 (w:w), or about 16.7:1 (w:w); and / or albumin:cholesterol is about (5-10):1 (w:w), e.g., about (6-9):1 (w:w), about (7-8):1 (w:w), about 10:1 (w:w), or about 6.7:1 (w:w); and / or The above composition is provided, wherein the ratio of Span 20:SN-38 is about (5-40):100 (w:w), for example, about (6-30):100 (w:w), about (7-25):100 (w:w), about (8-20):100 (w:w), about (9-15):100 (w:w), or about (10-12):100 (w:w).

[0096] In some embodiments, the SN-38 content is about 3 w / w% to about 9 w / w%, e.g., about 3.5 w / w% to about 8.5 w / w%, about 4 w / w%, about 4.5 w / w%, about 5 w / w%, about 5.5 w / w%, about 6 w / w%, about 6.5 w / w%, about 7 w / w%, about 7.5 w / w%, or about 8 w / w%, relative to the total amount of SN-38, cholesterol, and albumin in the composition.

[0097] In some embodiments, the cholesterol content is about 8 w / w% to about 18 w / w%, e.g., about 8.5 w / w% to about 17.5 w / w%, about 9 w / w%, about 9.5 w / w%, about 10 w / w%, about 10.5 w / w%, about 11 w / w%, about 11.5 w / w%, about 12 w / w%, about 12.5 w / w%, about 13 w / w%, about 13.5 w / w%, about 14 w / w%, about 14.5 w / w%, about 15 w / w%, about 15.5 w / w%, about 16 w / w%, about 16.5 w / w%, or about 17 w / w% of the total amount of SN-38, cholesterol, and albumin in the composition.

[0098] In some embodiments, the content of Span 20 relative to the total amount of SN-38, lipid, albumin, and Span 20 in the composition is about 0.2 w / w% to about 0.6 w / w%, e.g., about 0.22 w / w% to about 0.58 w / w%, about 0.24 w / w% to about 0.56 w / w%, about 0.26 w / w% to about 0.54 w / w%, about 0.28 w / w% to about 0.52 w / w%, about 0.3 w / w% to about 0.5 w / w%, about 0.32 w / w% to about 0.48 w / w%, about 0.34 w / w% to about 0.46 w / w%, about 0.36 w / w% to about 0.44 w / w%, about 0.38 w / w% to about 0.42 w / w%, or about 0.4 w / w%.

[0099] In some other further embodiments, the present invention provides cholesterol:SN-38 is about (0.8-1.8):1 (w:w), about (0.9-1.7):1 (w:w), or about (1-1.4):1; and / or albumin:SN-38 is about (9-21):1 (w:w), about (9-20):1 (w:w), about (11-18):1 (w:w), or about (11.1-17.3):1 (w:w); and / or albumin:cholesterol is about (6-21):1 (w:w), about (6.7-13):1 (w:w), about (7-13):1 (w:w), or about (11-12.7):1 (w:w); and / or Span 20:SN-38 is about (5-10):100(w:w), about (5-9):100(w:w), about (6-8.6):100(w:w), about (6-8):100(w:w), or about (6.5-7):100(w:w); and / or With respect to the total amount of SN-38, cholesterol, and albumin in the composition, the SN-38 content is about 4 w / w% to about 10 w / w%, about 4.5 w / w% to about 9.5 w / w%, about 5 w / w% to about 9 w / w%, or about 7.5 w / w% to about 8 w / w%; and / or a cholesterol content of about 4 w / w% to about 12.5 w / w%, about 4.5 w / w% to about 12 w / w%, about 7 w / w% to about 10 w / w%, or about 7.5 w / w% to about 8 w / w%; and / or an albumin content of about 78 w / w% to about 93 w / w%, about 79 w / w% to about 91.5 w / w%, about 80 w / w% to about 90 w / w%, about 82 w / w% to about 89 w / w%, about 84 w / w% to about 88 w / w%, or about 84.5 w / w% to about 87.5 w / w%; and / or The above composition is provided, wherein the content of Span 20 is about 0.2 w / w% to about 0.8 w / w%, about 0.24 w / w% to about 0.7 w / w%, about 0.26 w / w% to about 0.7 w / w%, about 0.3 w / w% to about 0.65 w / w%, about 0.36 w / w% to about 0.6 w / w%, about 0.4 w / w% to about 0.58 w / w%, about 0.44 w / w% to about 0.56 w / w%, about 0.48 w / w% to about 0.54 w / w%, or about 0.5 w / w% to about 0.52 w / w%.

[0100] Compositions according to the second subset exhibit advantages in scaled-up processes, e.g., in relatively large-scale preparations (e.g., hundreds of milligrams or more of SN-38 raw material), including pilot-scale preparations, such as reduced high-pressure homogenization during preparation, substantially reduced particle size of the nanoparticles in the composition, increased filtration flux, controllable particle size after disintegration under physiological conditions, reduced raw material loss and costs, and at the same time, suitable albumin content so that the particle size of the nanoparticles can be controlled to approach a particle size suitable for drug formulation.

[0101] In some embodiments of the first and second subsets, the SN-38 present in the nanoparticles comprises at least about 6 wt% to about 12 wt% of the total amount of SN-38, cholesterol, and albumin in the composition, e.g., about 7 wt% to about 11 wt%; about 8 wt% to about 10 wt%; about 8.3%; or about 9 wt%.

[0102] In some embodiments, the SN-38 present in the nanoparticles comprises about 95% to about 99% w / w of the total amount of SN-38 in the composition, e.g., about 96% to about 99% w / w, about 97% to about 99% w / w, about 98% to about 99% w / w, or greater than about 99% w / w.

[0103] In some of the above embodiments, the composition is in liquid, semi-solid, or solid form.

[0104] In some embodiments, the composition is in solid form, preferably in powder form, more preferably a lyophilized powder.

[0105] In some of these embodiments, the SN-38 is present in the composition preferably in amorphous and / or nanocrystalline form as determined by electron microscopy and X-ray diffraction (Cu-Kα) analysis.

[0106] In some embodiments, the composition does not contain an additional stabilizer. In some embodiments, the composition further comprises an additional stabilizer, wherein the additional stabilizer is present in an amount such that when the composition is reconstituted to form an aqueous composition (including solutions and emulsions), the content of the additional stabilizer is at least about 2 w / v%, e.g., at least about 3 w / v%, e.g., at least about 5 w / v%, about 5 w / v% to about 30 w / v%, about 10 w / v% to about 25 w / v%, or about 15 w / v% to about 20 w / v%.

[0107] In some of these embodiments, the composition further comprises an additional stabilizer in an amount of about 70% to about 96% by weight of the total composition, e.g., about 70% to about 90% by weight, about 72% to about 89% by weight, about 74% to about 88% by weight, about 76% to about 87% by weight, about 80% to about 86% by weight, about 81% to about 86% by weight, about 82% to about 85% by weight, or about 83% to about 84% by weight, based on the total amount of the composition. In some other embodiments, the composition further comprises an additional stabilizer in an amount of about 80% to about 96% by weight or about 84% to about 95% by weight of the total amount of the composition.

[0108] The additional stabilizer may be selected from mannitol, lactose, maltose, trehalose, dextran, glucose, and sucrose, and any compositions thereof, preferably sucrose.

[0109] In some other embodiments, the composition is an aqueous composition in liquid form, including solutions and emulsions.

[0110] In some of these embodiments, the composition in liquid form comprises SN-38 present in the form of nanocrystals and / or vesicles.

[0111] In some of these embodiments, the composition is in the form of a solution; in some embodiments, the composition is in the form of an emulsion.

[0112] In some of these embodiments, the amount of SN-38 is about 500.0 μg / mL to about 1.0 mg / mL, for example, about 600 μg / mL to about 800 μg / mL, relative to the total volume of the composition.

[0113] In some of these embodiments, the albumin content is about 5.0 mg / mL to about 10.0 mg / mL, for example, about 6.0 mg / mL to about 10 mg / mL, or about 7.0 mg / mL to about 8.0 mg / mL, relative to the total volume of the composition.

[0114] In some embodiments, the nanoparticles in the composition have an average particle size of about 90-160 nm, e.g., about 95-150 nm, about 100-140 nm, about 105-130 nm, about 110-125 nm, about 110 nm, about 115 nm, about 120 nm, about 125 nm, about 130 nm, 135 nm, about 140 nm, or about 145 nm.

[0115] In some embodiments, the nanoparticles have a particle size distribution index (PDI) of about 0.30 or less, eg, about 0.2 or less, about 0.10 or less, or about 0.01 or less.

[0116] In some embodiments, the composition has a zeta potential of about −35 mV to about −20 mV, for example, about −31 mV.

[0117] In some embodiments, the nanoparticles do not disintegrate when the composition is diluted (e.g., using 1x PBS at a pH of about 7.4) to result in an SN-38 content of about 4 μg / mL or less, e.g., about 2 μg / mL or less, about 1 μg / mL or less, about 0.4 μg / mL or less, about 0.1 μg / mL or less, about 0.04 μg / mL or less, about 0.02 μg / mL or less, or about 0.01 μg / mL or less in the diluted composition.

[0118] In some of these embodiments, the composition does not include an additional stabilizer. In some embodiments, the composition further includes an additional stabilizer, and the content of the additional stabilizer relative to the total weight of the composition is at least about 2 w / v%, e.g., at least about 3 w / v%, at least about 5 w / v%, about 5 w / v% to about 30 w / v%, about 10 w / v% to about 25 w / v%, or about 15 w / v% to about 20 w / v%.

[0119] The additional stabilizer may be selected from mannitol, lactose, maltose, trehalose, dextran, glucose, and sucrose, and any compositions thereof, preferably sucrose.

[0120] Ring-opened SN-38 is an inactive form of SN-38. The present inventors have surprisingly discovered that only small amounts of ring-opened SN-38 are present in the compositions of the present application. In some embodiments, the ring-opened SN-38 in the composition accounts for about 2 wt% or less, preferably about 1.8 wt% or less of the total amount of SN-38.

[0121] Albumin acting as a carrier may form multimers, including dimers, trimers, and multimers. The presence of albumin multimers increases the risk of immunogenicity of drugs, especially parenterally administered drugs. Therefore, it is advantageous to minimize the amount of albumin multimers present. The inventors surprisingly discovered that albumin multimers are absent or substantially absent from the compositions of the present application. Preferably, the monomeric form of albumin in the composition accounts for at least about 95% w / w, preferably at least about 96%, more preferably at least about 98%, more preferably at least about 99%, at least about 99.2%, at least about 99.4%, or at least about 99.5% of the total amount of albumin. The compositions of the present application are expected to exhibit the advantage of low immunogenicity and therefore high safety.

[0122] The albumin that can be used in the present application is selected from human serum albumin (HSA), recombinant human albumin (rHA), bovine serum albumin, and porcine serum albumin. For example, the albumin comprises the amino acid sequence shown in SEQ ID NO: 1. Preferably, the albumin is selected from human serum albumin (HSA) and recombinant human albumin (rHA).

[0123] In a second aspect, the present application provides a method for manufacturing a semiconductor device comprising: (1) dissolving SN-38, lipid, and Span 20 in an organic solvent to form an organic phase; and preparing an aqueous solution of albumin as an aqueous phase; (2) mixing an organic phase with an aqueous phase to form an emulsion, the emulsion comprising nanoparticles in which albumin encapsulates at least a portion of the SN-38 and optionally at least a portion of the lipid; and (3) Removing the organic solvent from the emulsion to obtain a product containing nanoparticles. The present invention provides a method for preparing a composition according to the first aspect, comprising:

[0124] In some embodiments, the method comprises: (1) SN-38, lipids, and Span 20 were mixed in DMSO and C 1~3 dissolving the albumin using a mixed organic solvent, the mixed organic solvent comprising a first organic solvent selected from an alcohol and a second organic solvent selected from CHCl3 and a mixture of CH2Cl2 and CHCl3, to form an organic phase; preparing an aqueous solution of albumin as an aqueous phase; (2) mixing an organic phase with an aqueous phase to prepare an emulsion, the emulsion comprising nanoparticles in which albumin encapsulates at least a portion of the SN-38 and optionally at least a portion of the lipid; (3) removing the organic solvent; and (4) Optionally, sterilizing the product obtained in step (3).

[0125] In some preferred embodiments, in the mixed organic solvent in step (1), the second organic solvent is DMSO or C 1~3 The volume ratio of alcohols is about 1:20 (v / v) to about 20:1 (v / v), for example, about 1:5 to about 5:1 (v / v), about 1:2 to about 4:1 (v / v), about 1:1 to about 4:1 (v / v), about 1.5:1 (v / v) to about 3:1 (v / v), or about 2:1 (v / v) to 7:3 (v / v).

[0126] In some preferred embodiments, in step (2), the ratio of organic phase to aqueous phase is about 1:2 (v / v) to about 1:50 (v / v), for example, about 1:5 (v / v) to about 1:20 (v / v), about 1:7 (v / v) to about 1:15 (v / v), or 1:10 (v / v) to about 1:12 (v / v), for example, about 1:5 (v / v) to about 1:12 (v / v), about 1:5 (v / v) to about 1:12 (v / v), about 1:6 (v / v), about 1:7 (v / v), or about 1:10 (v / v).

[0127] In some preferred embodiments, step (2) (2-1) dispersing an organic phase in an aqueous phase under shear to obtain a crude emulsion; and (2-2) Homogenizing the coarse emulsion under high pressure to obtain a fine emulsion containing nanoparticles Includes:

[0128] C 1~3 Alcohols include methanol, ethanol, and isopropanol, and any combination thereof, for example, ethanol (EtOH).

[0129] In some embodiments, the mixed organic solvent comprises the second organic solvent and EtOH at about 7:3 (v / v), hi some embodiments, the mixed organic solvent comprises the second organic solvent and DMSO at about 1:1 (v / v).

[0130] In some further embodiments, the present invention provides (1) dissolving SN-38, lipid, and Span 20 in a 1:1 (v / v) mixed organic solvent of the second organic solvent / DMSO or a 7:3 (v / v) mixed organic solvent of the second organic solvent / EtOH to form an organic phase; preparing an aqueous solution of albumin as an aqueous phase; (2) mixing an organic phase and an aqueous phase in a ratio of about 1:10 (v / v) to about 1:15 (v / v), for example about 1:12 (v / v), to prepare an emulsion, the emulsion comprising nanoparticles in which albumin encapsulates at least a portion of the SN-38 and, optionally, at least a portion of the lipid; (3) removing the organic solvent; and (4) optionally, providing the above method, which comprises a step of sterilizing the product obtained in step (3).

[0131] In some embodiments, the second organic solvent is CHCl. In some other embodiments, the second organic solvent is a mixture of CHCl and CHCl, preferably in a volume ratio of about 2:5 to 1:1, preferably about 2:5, of CHCl to CHCl in the mixture. The use of a mixture of CHCl and CHCl as the second organic solvent has the advantage of reducing the residual CHCl level in the final product compared to CHCl alone, thus reducing residual solvent limitations during clinical administration.

[0132] In some embodiments, the aqueous phase does not contain any additional stabilizers.

[0133] In some other embodiments, the aqueous phase already contains an additional stabilizer. In some other embodiments, the method further comprises adding an additional stabilizer in step (2). For example, the additional stabilizer is present in an amount such that the content of the additional stabilizer in the product obtained in step (3) or (4) is at least about 2 w / v%, e.g., at least about 3 w / v%, at least about 5 w / v%, about 5 w / v% to about 30 w / v%, about 10 w / v% to about 25 w / v%, or about 15 w / v% to about 20 w / v%. Preferably, the additional stabilizer can be selected from albumin (e.g., human serum albumin, recombinant human albumin, bovine serum albumin, and skim milk powder), monosaccharides, disaccharides, polysaccharides, mannitol, and any combination thereof, preferably mannitol, lactose, maltose, trehalose, dextran, glucose, and sucrose, and any composition thereof, preferably sucrose.

[0134] In some embodiments, the organic solvent mixture in step (1) is added to the aqueous phase before mixing the organic phase and the aqueous phase in step (2). For example, the volume of the added organic solvent mixture is equal to or less than the volume of the organic phase. For example, the volume ratio of the added organic solvent mixture to the organic phase is about 1:1 (v / v) to about 1:5 (v / v), e.g., about 1:2 (v / v) to about 1:4 (v / v), or about 1:3 (v / v).

[0135] In some embodiments, the concentration of SN-38 in the organic phase in step (1) is about 5 to 17 mg / mL, for example, about 5.25 to 12 mg / mL or about 7 to 12 mg / mL, for example, about 10 mg / mL.

[0136] In some other embodiments, the concentration of SN-38 in the organic phase in step (1) is 4-10 mg / mL, for example, about 6-8 mg / mL.

[0137] In some embodiments, the concentration of lipid in the organic phase in step (1) is about 3-50 mg / mL, e.g., about 5-45 mg / mL, or about 7.5-30 mg / mL, about 10-25 mg / mL, or about 15-20 mg / mL.

[0138] In some other embodiments, the concentration of lipid in the organic phase in step (1) is about 10-20 mg / mL, for example, about 15 mg / mL.

[0139] In some embodiments, the concentration of Span 20 in the organic phase in step (1) is about 0.3 to 6 mg / mL, for example, about 0.3 to 2 mg / mL or about 0.6 to 1 mg / mL.

[0140] In some embodiments, the concentration of albumin in the aqueous phase is about 5 to 15 mg / mL, for example, about 6 to 12 mg / mL, preferably about 6 to 10 mg / mL.

[0141] In some other embodiments, the concentration of albumin in the aqueous phase is about 8-30 mg / mL, such as about 12-20 mg / mL or about 16-18 mg / mL.

[0142] In some embodiments, the method includes a step (4) of sterilizing the product obtained in step (3). The sterilization method is not particularly limited. In a preferred embodiment, the product obtained in step (3) is sterilized by filtration through a filter membrane of about 0.2 μm.

[0143] In some embodiments, the method comprises: (5) The method further comprises the step of drying the product obtained in step (3) or (4), preferably by spray drying or freeze drying, to obtain a solid form of the composition, preferably a powder, more preferably a freeze-dried powder.

[0144] Preferably, the SN-38 is present in the composition in amorphous and / or nanocrystalline form as determined by electron microscopy or X-ray diffraction (Cu-Kα) analysis.

[0145] In some further embodiments, step (5) further comprises adding an additional stabilizer to the product obtained in step (3) or (4) prior to drying, wherein the additional stabilizer is present in an amount such that when the solid form obtained in step (5) is reconstituted to form an aqueous composition (including solutions and emulsions), the additional stabilizer content is at least about 2 w / v%, e.g., at least about 3 w / v%, at least about 5 w / v%, between about 5 w / v% and about 30 w / v%, between about 10 w / v% and about 25 w / v%, or between about 15 w / v% and about 20 w / v%.

[0146] The additional stabilizer may be selected from albumin (e.g., human serum albumin, recombinant human albumin, bovine serum albumin, and skim milk powder), monosaccharides, disaccharides, polysaccharides, mannitol, and any combination thereof, preferably mannitol, lactose, maltose, trehalose, dextran, glucose, and sucrose, and any composition thereof, preferably sucrose.

[0147] In a third aspect, the present application provides a composition comprising SN-38, a lipid, albumin, and Span 20, (1) dissolving SN-38, lipid, and Span 20 in an organic solvent to form an organic phase; and preparing an aqueous solution of albumin as an aqueous phase; (2) mixing an organic phase with an aqueous phase to form an emulsion, the emulsion comprising nanoparticles in which albumin encapsulates at least a portion of the SN-38 and optionally at least a portion of the lipid; and (3) Removing the organic solvent from the emulsion to obtain a product containing nanoparticles. The present invention provides a method for preparing a composition, comprising:

[0148] Embodiments of the method are as described above for the method according to the second aspect.

[0149] In the methods of the second and third aspects, the portion of albumin may encapsulate a portion of SN-38 to form a nanoparticle, or may encapsulate a portion of lipid to form a nanoparticle. Thus, in some embodiments, the compositions of the first and fourth aspects below may comprise nanoparticles formed by a portion of albumin encapsulating a portion of SN-38 and / or a portion of albumin encapsulating a portion of lipid.

[0150] In a fourth aspect, the present application provides a composition preparable by the method according to the third aspect above.

[0151] In a fifth aspect, the present application further provides a composition comprising SN-38, a lipid, and albumin, wherein the albumin encapsulates at least a portion of the SN-38 and optionally at least a portion of the lipid to form nanoparticles;

[0152] A method is provided for preparing a composition with improved properties, characterized in that Span 20 is added during the preparation of the composition.

[0153] In some embodiments, the composition does not include an added stabilizer.

[0154] In some embodiments, the improved properties include improved stability.

[0155] In some embodiments, when the composition is in liquid form, improved stability comprises a reduction in the formation or content of albumin multimers and / or a reduction in nanoparticle size during preparation, storage, and / or use of the composition. Preferably, albumin multimers are absent or substantially absent from the composition, or albumin multimers account for at most 5% w / w of the total amount of albumin, e.g., at most about 4%, at most about 2%, at most about 1.5%, at most about 1.2%, at most about 1.1%, at most about 1%, or at most about 0.8%.

[0156] In some embodiments, the composition is as described in the first aspect above.

[0157] In some embodiments, the method comprises: (1) dissolving SN-38, lipid, and Span 20 in an organic solvent to form an organic phase; and preparing an aqueous solution of albumin as an aqueous phase; (2) mixing an organic phase with an aqueous phase to form an emulsion, the emulsion comprising nanoparticles in which albumin encapsulates at least a portion of the SN-38 and optionally at least a portion of the lipid; and (3) Removing the organic solvent from the emulsion to obtain a product containing nanoparticles. Includes:

[0158] Further embodiments of the method are as described above for the method according to the second aspect.

[0159] In a sixth aspect, the present application provides a pharmaceutical composition comprising the above composition and, optionally, a pharmaceutically acceptable carrier.

[0160] In a seventh aspect, the present application provides a pharmaceutical composition comprising the above-described composition in a dried state and, optionally, a pharmaceutically acceptable carrier. The drying is preferably lyophilization or spray drying, more preferably lyophilization. Preferably, the pharmaceutical composition is in a solid form, preferably a lyophilized powder. Preferably, SN-38 is present in the composition in an amorphous and / or nanocrystalline form.

[0161] There is no particular limitation on the route of administration of the compositions or pharmaceutical compositions of the present application. Possible routes of administration include, but are not limited to, oral administration, intranasal administration, topical administration, and parenteral administration. Preferably, the pharmaceutical composition is used for parenteral administration, including, but not limited to, intravenous administration, intraarterial administration, subcutaneous administration, intradermal administration, and intramuscular administration, more preferably, administration by intravenous injection (e.g., bolus or infusion).

[0162] The selection of a pharmaceutically acceptable carrier depends on the dosage form of the drug or pharmaceutical composition, primarily on the route of administration of the dosage form (e.g., oral, intranasal, intradermal, subcutaneous, topical, intramuscular, or intravenous administration), and secondarily on the formulation of the dosage form. For example, pharmaceutically acceptable carriers include water (e.g., water for injection), buffer solutions, isotonic saline (e.g., phosphate buffered saline (PBS)), glucose, mannitol, dextrose, lactose, starch, magnesium stearate, cellulose, magnesium carbonate, 0.3% glycerin, hyaluronic acid, ascorbic acid, lactic acid, ethanol, polyalkylene glycols, such as polyethylene glycol (e.g., polyethylene glycol 4000) or polypropylene glycol, triglycerides, etc.

[0163] In an eighth aspect, the present application provides the use of the composition or pharmaceutical composition described above in the manufacture of a medicament for treating an SN-38-sensitive tumor in a subject.

[0164] In a ninth aspect, the present application provides a composition or pharmaceutical composition as described above for use in treating an SN-38-sensitive tumor in a subject.

[0165] In a tenth aspect, the present application provides a method for treating an SN-38-sensitive tumor in a subject, comprising administering to the subject a therapeutically effective amount of the composition or pharmaceutical composition described above.

[0166] In some embodiments, the subject is a mammal, including but not limited to, a mouse, rat, rabbit, guinea pig, dog, cat, sheep, cow, goat, and horse, hi some embodiments, the individual is a human.

[0167] "SN-38-sensitive tumor" refers to a tumor that is responsive to administration of SN-38, and responses include a reduction in tumor cells, a reduction in tumor diameter, a elimination of tumor metastasis, an inhibition of tumor growth, etc. Preferably, the SN-38-sensitive tumor is selected from colorectal cancer, small cell lung cancer, lymphoma, breast cancer (preferably triple-negative breast cancer), esophageal cancer, gastric cancer, liver cancer, renal cancer, pancreatic cancer, uterine cancer, and ovarian cancer.

[0168] Experimental studies have shown that the antitumor effects of antitumor drugs can be significantly improved after they are bound to albumin. For example, nanoalbumin-bound paclitaxel has tumor tissue selectivity and a unique transport mechanism, increasing intratumoral paclitaxel concentrations by 33% compared to paclitaxel in solution (Desai N, Trieu V, Yao Z, et al., Increased antitumor activity, intratumor paclitaxel concentrations, and endothelial cell transport of cremophor-free, albumin-bound paclitaxel, ABI-007, compared with cremophor-based paclitaxel, [J]. Clinical cancer research, 2006, 12(4):1317-1324). The above results are due to the active tumor targeting effect achieved by the binding of the drug to albumin followed by the gp60 receptor in tumor tissue, and the passive tumor targeting effect achieved by the EPR effect of the drug-albumin conjugate nanoparticles. Therefore, drug distribution in the tumor is significantly increased, improving therapeutic efficacy and reducing toxic and side effects. The compositions or pharmaceutical compositions of the present application are expected to exhibit advantageous antitumor efficacy. Through preliminary experiments, the inventors have found that the compositions of the present application exhibit superior antitumor activity compared to that of commercially available irinotecan hydrochloride injection.

[0169] In an eleventh aspect, the present application provides a kit comprising the composition or pharmaceutical composition described above. If necessary, the kit may further comprise instructions, packaging, and a container for housing the composition or pharmaceutical composition.

[0170] Although the embodiments of the present invention have been described in the above first to eleventh aspects using SN-38 as the active ingredient, these are merely one aspect of the inventive concept. The inventive concept further contemplates technical solutions using other camptothecin drugs as the active ingredient. It is expected that the above embodiments will still be applicable when other camptothecin drugs are used instead of SN-38. In other words, the present application further includes, unless the context requires otherwise, any embodiment as described in any one of the above first to eleventh aspects, except that other camptothecin drugs are used instead of SN-38. Other camptothecin drugs include irinotecan (CPT-11), 10-hydroxycamptothecin (HCPT), topotecan (TPT), rubitecan (9-NC), 9-aminocamptothecin (9-AC), belotecan (Cas. No.: 256411-32-2), Dxd (Cas. No.: 1599440-33-1), and DX-8951 (exa The agonist may be selected from the group consisting of CKD602 (belotecan), lurtotecan, namitecan (Cas. No.: 372105-27-6), ST1481 (gimatecan, Cas. No.: 292618-32-7), BNP-1350 (Cas. No. 203923-89-1), and BN80915 (diflomotecan). Those skilled in the art will recognize that these embodiments can also achieve the beneficial effects described below.

[0171] Embodiments of the invention described in the "Summary of the Invention" section of this application include: Embodiment 1 A composition comprising SN-38, a lipid, albumin, and Span 20, wherein the composition comprises nanoparticles, wherein the albumin encapsulates at least a portion of the SN-38 and optionally at least a portion of the lipid; lipid:SN-38 is about (0.1-10):1 (w:w), about (0.5-6):1 (w:w), about (0.5-5):1 (w:w), about (0.5-3):1 (w:w), about (1-4):1 (w:w), about (1.2-4):1 (w:w), about (1.4-2):1 (w:w), about (1.5-2.5):1 (w:w), or about 1:1; albumin:SN-38 is about (1-100):1 (w:w), about (1-50):1 (w:w), about (3-25):1 (w:w), about (5-25):1 (w:w), about (5-20):1 (w:w), about (5-18):1 (w:w), about (6-15):1 (w:w), about (7-15):1 (w:w), about (6-12):1 (w:w), about (7-12):1 (w:w), about (9-11):1 (w:w), or about 10:1 (w:w); Span 20:SN-38 is about (3-60):100(w:w), about (4-60):100(w:w), about (5-60):100(w:w), about (6-60):100(w:w), about (7-55):100(w:w), about (8-50):100(w:w), about (10-45):100(w:w), about (12-40):100(w:w), about (14-35):100(w:w), about (15-30):100(w:w), about (16-25):100(w:w), or about (18-20):100(w:w); A composition characterized in that the lipid is selected from cholesterol, cholesterol derivatives, cholesterol analogs, and fatty acid esters, and any combination of two or more thereof.

[0172] Embodiment 2 2. The composition of embodiment 1, wherein the albumin:lipid ratio is about (1-100):1 (w:w), such as about (2-20):1 (w:w), about (3-15):1 (w:w), about (5-10):1 (w:w), about 7:1 (w:w), or about 10:1 (w:w).

[0173] Embodiment 3 With respect to the total amount of SN-38, lipid, and albumin in the composition, an SN-38 content of about 1% w / w to about 25% w / w; and / or a lipid content of about 1% w / w to about 35% w / w; and / or The albumin content is about 50 w / w% to about 98 w / w%; Or, The content of SN-38 is about 3% w / w to about 20% w / w; and / or a lipid content of about 2% w / w to about 30% w / w; and / or The albumin content is about 55 w / w% to about 95 w / w%; Or, an SN-38 content of about 3 w / w% to about 15 w / w%, about 4 w / w%, about 5 w / w%, about 6 w / w%, about 6.5 w / w%, about 7 w / w%, about 7.5 w / w%, about 8 w / w%, about 8.5 w / w%, about 9 w / w%, about 10 w / w%, about 11 w / w%, about 12 w / w%, about 13 w / w%, or about 14 w / w%; and / or a lipid content of about 3 w / w% to about 30 w / w%, about 4 w / w%, about 5 w / w%, about 6 w / w%, about 7 w / w%, about 8 w / w%, about 8.5 w / w%, about 9 w / w%, about 9.5 w / w%, about 10 w / w%, about 10.5 w / w%, about 11 w / w%, about 11.5 w / w%, about 12 w / w%, about 12.5 w / w%, about 13 w / w%, about 13.5 w / w%, about 14 w / w%, about 15 w / w%, about 16 w / w%, about 17 w / w%, about 18 w / w%, about 19 w / w%, about 20 w / w%, about 21 w / w%, about 24 w / w%, about 26 w / w%, or about 28 w / w%; and / or The composition according to embodiment 1 or 2, characterized in that the albumin content is about 60 w / w% to about 94 w / w%, about 64 w / w% to about 93 w / w%, about 66 w / w% to about 92 w / w%, about 68 w / w% to about 91 w / w%, about 70 w / w% to about 90 w / w%, about 75 w / w% to about 90 w / w%, about 76 w / w%, about 77 w / w%, about 78 w / w%, about 79 w / w%, about 80 w / w%, about 81 w / w%, about 82 w / w%, about 83 w / w%, about 84 w / w%, about 85 w / w%, about 86 w / w%, about 87 w / w%, about 88 w / w%, or about 89 w / w%.

[0174] Embodiment 4 4. The composition according to any one of embodiments 1 to 3, wherein the content of Span 20 is about 0.03 w / w% to about 12 w / w%, about 0.06 w / w% to about 10 w / w%, 0.08 w / w% to about 9 w / w%, about 0.1 w / w% to about 8 w / w%, about 0.2 w / w% to about 7 w / w%, about 0.4 w / w% to about 6 w / w%, about 0.6 w / w% to about 5 w / w%, about 0.8 w / w% to about 4 w / w%, or about 1 w / w% to about 2 w / w% relative to the total amount of SN-38, lipid, albumin, and Span 20 in the composition.

[0175] Embodiment 5 Lipid:SN-38 is approximately (1-10):1(w:w), approximately (1-8):1(w:w), approximately (1-6):1(w:w), approximately (1-5):1(w:w), approximately (1-4.5):1(w:w), approximately (1-4):1(w:w), approximately (1.2-3.8):1(w:w), approximately (1.4-3.6):1(w:w), approximately (1.6-3.4):1(w:w), approximately (1.8-3.2):1(w:w), about (2-3):1(w:w), about (2.2-2.8):1(w:w), about (2.4-2.6):1(w:w), about 1:1(w:w), about 2.5:1(w:w), or about (0.8-1.8):1(w:w), about (0.9-1.7):1(w:w), or about (1-1.4):1; and / or albumin:SN-38 is about (5-40):1(w:w), about (less than 5-40):1(w:w), about (5-35):1(w:w), about (5-30):1(w:w), about (8-25):1(w:w), about (10-22.5):1(w:w), about (12.5-20):1(w:w), about (15-17.5):1(w:w), about (16-18):1(w:w), about 10:1(w:w), or about (9-21):1(w:w), about (9-20):1(w:w), about (11-18):1(w:w), or about (11.1-17.3):1(w:w); and / or Span 20:SN-38 is approximately (5-60): 100 (w:w), approximately (6-60): 100 (w:w), approximately (7-55): 100 (w:w), approximately (8-50): 100 (w:w), approximately (10-45): 100 (w:w), approximately (12-40): 100 (w:w), approximately (14-35): 100 (w:w), approximately (15-30): 100 (w:w), about (16-25):100(w:w), about (18-20):100(w:w), or about (5-10):100(w:w), about (5-9):100(w:w), about (6-8.6):100(w:w), about (6-8):100(w:w), or about (6.5-7):100(w:w); and / or 5. The composition according to any one of embodiments 1 to 4, wherein the albumin:lipid ratio is about (1 to 40):1 (w:w), about (less than 1 to 40):1 (w:w), about (2 to 35):1 (w:w), about (3 to 15):1 (w:w), about (5 to 10):1 (w:w), about (6 to 8):1 (w:w), or about 7:1 (w:w), or about (6 to 21):1 (w:w), about (6.7 to 13):1 (w:w), about (7 to 13):1 (w:w), or about (11 to 12.7):1 (w:w).

[0176] Embodiment 6 With respect to the total amount of SN-38, lipid, and albumin in the composition, an SN-38 content of about 2 wt% to about 16 wt%; and / or a lipid content of about 2% w / w to about 35% w / w; and / or The albumin content is about 75 w / w% to about 96 w / w%; Or, the SN-38 content is about 2.5 w / w% to about 15 w / w%, about 4 w / w% to about 10 w / w%, about 4.5 w / w% to about 9.5 w / w%, about 5 w / w% to about 9 w / w%, or about 7.5 w / w% to about 8 w / w%; and / or The lipid content in the composition is about 2.5 w / w% to about 30 w / w%, about 4 w / w% to about 12.5 w / w%, about 4.5 w / w% to about 12 w / w%, about 7 w / w% to about 10 w / w%, or about 7.5 w / w% to about 8 w / w%; and / or the albumin content in the composition is about 76 w / w% to about 95 w / w%, about 78 w / w% to about 93 w / w%, about 79 w / w% to about 91.5 w / w%, about 80 w / w% to about 90 w / w%, about 82 w / w% to about 89 w / w%, about 84 w / w% to about 88 w / w%, or about 84.5 w / w% to about 87.5 w / w%; Or, The SN-38 content is approximately 3 w / w% to approximately 14 w / w%, approximately 3.5 w / w% to approximately 12 w / w%, approximately 4 w / w%, approximately 4.2 w / w%, approximately 4.5 w / w%, approximately 4.6 w / w%, approximately 4.8 w / w%, approximately 5 w / w%, approximately 5.5 w / w%, approximately 6 w / w%, approximately 6.5 w / w%, approximately 7 w / w%, approximately 7.5 w / w / w%, about 7.6 w / w%, about 7.8 w / w%, about 8 w / w%, about 8.5 w / w%, about 9 w / w%, about 9.2 w / w%, about 9.5 w / w%, about 9.6 w / w%, about 9.8 w / w%, about 10 w / w%, about 10.5 w / w%, about 11 w / w%, or about 11.5 w / w%; and / or Lipid content is about 4 w / w% to about 25 w / w%, about 4.5 w / w% to about 20 w / w%, about 4.3 w / w%, about 4.5 w / w%, about 4.7 w / w%, about 5 w / w%, about 5.5 w / w%, about 6 w / w%, about 6.5 w / w%, about 6.7 w / w%, about 6.9 w / w%, about 7 w / w%, about 7.5 w / w%, about 7.6 w / w%, about 7.8 w / w%, about 8 w / w%, about 8.5 w / w%, about 9 w / w%, about 9.5 w / w%, about 10 w / w%, about 10.5 w / %, about 11 w / w%, about 11.5 w / w%, about 12 w / w%, about 12.1 w / w%, about 12.3 w / w%, about 12.5 w / w%, about 13 w / w%, about 13.5 w / w%, about 14 w / w%, about 14.5 w / w%, about 15 w / w%, about 15.5 w / w%, about 16 w / w%, about 16.5 w / w%, about 17 w / w%, about 17.5 w / w%, about 18 w / w%, about 18.5 w / w%, about 19 w / w%, or about 19.5 w / w%; and / or Albumin content: approximately 78 w / w% to approximately 92 w / w%, approximately 79 w / w%, approximately 79.2 w / w%, approximately 79.4 w / w%, approximately 79.6 w / w%, approximately 79.8 w / w%, approximately 80 w / w%, approximately 81 w / w%, approximately 82 w / w%, approximately 83 w / w%, approximately 84 w / w%, approximately 84.3 w / w%, approximately 84.5 w / w%, approximately 84.7 w / w%, approximately 84.9 w / w% 85 w / w%, about 86 w / w%, about 87 w / w%, about 87.3 w / w%, about 87.5 w / w%, about 87.7 w / w%, about 87.9 w / w%, about 88 w / w%, about 89 w / w%, about 90 w / w%, about 91 w / w%, about 91.3 w / w%, or about 91.5 w / w%.

[0177] Embodiment 7 The content of Span 20 relative to the total amount of SN-38, lipid, albumin, and Span 20 in the composition is about 0.14 w / w% to about 5 w / w%, about 0.2 w / w% to about 2.5 w / w%, about 0.22 w / w% to about 2.0 w / w%, about 0.24 w / w% to about 2 w / w%, about 0.26 w / w% to about 1.5 w / w%, about 0.28 w / w% to about 1.0 w / w%, about 0.3 w / w% to about 0 .9w / w%, about 0.32w / w% to about 0.8w / w%, about 0.34w / w% to about 0.7w / w%, about 0.36w / w% to about 0.6w / w% , about 0.38w / w%~about 0.58w / w%, about 0.4w / w%~about 0.56w / w%, about 0.42w / w%~about 0.54w / w%, about 0.4 7. The composition according to any one of embodiments 1 to 6, wherein the concentration is 4 w / w% to about 0.52 w / w%, about 0.46 w / w%, about 0.48 w / w%, about 0.5 w / w%, about 0.2 w / w% to about 0.8 w / w%, about 0.24 w / w% to about 0.7 w / w%, about 0.26 w / w% to about 0.7 w / w%, about 0.3 w / w% to about 0.65 w / w%, about 0.36 w / w% to about 0.6 w / w%, about 0.4 w / w% to about 0.58 w / w%, about 0.44 w / w% to about 0.56 w / w%, about 0.48 w / w% to about 0.54 w / w%, or about 0.5 w / w% to about 0.52 w / w%.

[0178] Embodiment 8 the SN-38 present in the nanoparticles comprises at least about 1 wt.% or at least about 2 wt.% of the total amount of SN-38, lipid, and albumin in the composition, e.g., at least about 3 wt.%, about 3 wt.% to about 13 wt.%, about 4 wt.% to about 12 wt.%, about 4 wt.%, about 5 wt.%, about 6 wt.%, about 7 wt.%, about 8 wt.%, about 9 wt.%, about 10 wt.%, or about 11 wt.%; and / or 8. The composition of any one of embodiments 1 to 7, wherein the SN-38 present in the nanoparticles accounts for about 80 w / w% to about 99 w / w% of the total amount of SN-38 in the composition, e.g., about 88 w / w% to about 98 w / w%, about 89 w / w%, about 90 w / w%, about 91 w / w%, about 92 w / w%, about 93 w / w%, about 94 w / w%, about 95 w / w%, about 96 w / w%, or about 97 w / w%.

[0179] Embodiment 9 the cholesterol derivative is selected from esters formed by cholesterol and organic acids, including cholesteryl palmitate, cholesteryl caprylate, and combinations thereof; and / or The cholesterol analog is selected from vitamin D2, vitamin D3, and combinations thereof; and / or 9. The composition according to any one of the preceding embodiments, wherein the fatty acid ester is selected from fatty acid glycerides, such as long chain fatty acid glycerides, including glyceryl stearate, such as glyceryl monostearate.

[0180] Embodiment 10 10. The composition according to any one of embodiments 1 to 9, wherein the lipid is cholesterol.

[0181] Embodiment 11 cholesterol:SN-38 is about (1-6):1 (w:w), about (1.2-5):1 (w:w), about (1.4-4):1 (w:w), about 3:1 (w:w), about 2:1 (w:w), about 1:1 (w:w), or about (0.8-1.8):1 (w:w), about (0.9-1.7):1 (w:w), or about (1-1.4):1; and / or albumin:SN-38 is about (3-25):1 (w:w), about (4-20):1 (w:w), about (5-15):1 (w:w), about (6-12):1 (w:w), about (7-12):1 (w:w), about (9-11):1 (w:w), or about 10:1 (w:w), or about (9-21):1 (w:w), about (9-20):1 (w:w), about (11-18):1 (w:w), or about (11.1-17.3):1 (w:w); and / or albumin:cholesterol is about (2-20):1 (w:w), about (3-15):1 (w:w), about (5-10):1 (w:w), or about 7:1 (w:w), or about (6-21):1 (w:w), about (6.7-13):1 (w:w), about (7-13):1 (w:w), or about (11-12.7):1 (w:w); and / or With respect to the total amount of SN-38, cholesterol, and albumin in the composition, the SN-38 content is about 3 w / w% to about 15 w / w%, about 4 w / w% to about 15 w / w%, about 6 w / w% to about 10 w / w%, or about 8 w / w% to about 12 w / w%, or about 4 w / w% to about 10 w / w%, about 4.5 w / w% to about 9.5 w / w%, about 5 w / w% to about 9 w / w%, or about 7.5 w / w% to about 8 w / w%; and / or a cholesterol content of about 5 w / w% to about 25 w / w%, about 6 w / w% to about 22 w / w%, about 15 w / w% to about 20 w / w%, or about 4 w / w% to about 12.5 w / w%, about 4.5 w / w% to about 12 w / w%, about 7 w / w% to about 10 w / w%, or about 7.5 w / w% to about 8 w / w%; and / or an albumin content of about 64 w / w% to about 90 w / w%, about 70 w / w% to about 90 w / w%, or about 78 w / w% to about 93 w / w%, about 79 w / w% to about 91.5 w / w%, about 80 w / w% to about 90 w / w%, about 82 w / w% to about 89 w / w%, about 84 w / w% to about 88 w / w%, or about 84.5 w / w% to about 87.5 w / w%; and / or 11. The composition of embodiment 10, wherein the SN-38 present in the nanoparticles comprises at least about 3 wt% of the total amount of SN-38, cholesterol, and albumin in the composition, e.g., about 3 wt% to about 13 wt%, about 4 wt% to about 12 wt%, about 4 wt%, about 5 wt%, about 6 wt%, about 7 wt%, about 8 wt%, about 9 wt%, about 10 wt%, or about 11 wt%.

[0182] Embodiment 12 cholesterol:SN-38 is about (1-3):1(w:w), about (1.2-2.5):1(w:w), about (1.4-2):1(w:w), about (1.5-2):1(w:w), about (1.3-1.8):1(w:w), about (1.4-1.6):1(w:w), about (1.5-1.7):1(w:w), about (1.2-1.5):1(w:w), about 1:1(w:w), about (1.4-1.5):1(w:w), or about (0.8-1.8):1(w:w), about (0.9-1.7):1(w:w), or about (1-1.4):1; albumin:SN-38 is about (5-15):1(w:w), about (5-12):1(w:w), about (6-12):1(w:w), or about (7-12):1(w:w), about (9-11):1(w:w), about (10-12):1(w:w), about 11:1(w:w), or about (9-21):1(w:w), about (9-20):1(w:w), about (11-18):1(w:w), or about (11.1-17.3):1(w:w); the albumin:cholesterol ratio is about (3-10):1 (w:w), about (4-8):1 (w:w), about (5-7):1 (w:w), or about (6-21):1 (w:w), about (6.7-13):1 (w:w), about (7-13):1 (w:w), or about (11-12.7):1 (w:w); 11. The composition of embodiment 10.

[0183] Embodiment 13 With respect to the total amount of SN-38, cholesterol, and albumin in the composition, the SN-38 content is about 6 w / w% to about 14 w / w%, about 6.5 w / w% to about 13 w / w%, about 7 w / w% to about 12 w / w%, about 7.5 w / w% to about 12 w / w%, about 8 w / w% to about 11 w / w%, about 8.5 w / w% to about 10 w / w%, about 9 w / w%, or about 4 w / w% to about 10 w / w%, about 4.5 w / w% to about 9.5 w / w%, about 5 w / w% to about 9 w / w%, or about 7.5 w / w% to about 8 w / w%; and / or Cholesterol content is about 8 w / w% to about 18 w / w%, about 8.5 w / w% to about 17 w / w%, about 9 w / w% to about 16 w / w%, about 9.5 w / w% to about 16 w / w%, about 10 w / w% to about 16 w / w%, about 10.5 w / w% to about 16 w / w%, about 11 w / w% to about 15 w / w%, about 11.5 w / w% to about 1 5 w / w%, about 12 w / w% to about 15 w / w%, about 12.5 w / w% to about 14 w / w%, about 13 w / w% to about 13.5 w / w%, or about 4 w / w% to about 12.5 w / w%, about 4.5 w / w% to about 12 w / w%, about 7 w / w% to about 10 w / w%, or about 7.5 w / w% to about 8 w / w%; and / or Albumin content is about 66 w / w% to about 90 w / w%, about 68 w / w% to about 89 w / w, about 70 w / w% to about 88 w / w%, about 70 w / w% to about 87 w / w%, about 70 w / w% to about 86 w / w%, about 70 w / w% to about 85 w / w%, about 75 w / w% to about 85 w / w%, about 76 w / w%, about 77 w / w%, about 78 w / w%, about 79 w / w%, 80 w / w%, 81 w / w%, about 82 w / w%, about 83 w / w%, about 84 w / w%, about 78 w / w% to about 93 w / w%, about 79 w / w% to about 91.5 w / w%, about 80 w / w% to about 90 w / w%, about 82 w / w% to about 89 w / w%, about 84 w / w% to about 88 w / w%, or about 84.5 w / w% to about 87.5 w / w%.

[0184] Embodiment 14 cholesterol:SN-38 is about (1-5):1(w:w), about (1-4.5):1(w:w), about (1-4):1(w:w), about (1.2-3.8):1(w:w), about (1.4-3.6):1(w:w), about (1.6-3.4):1(w:w), about (1.8-3.2):1(w:w), about (2-3):1(w:w), about (2.2-2.8):1(w:w), about (2.4-2.6):1(w:w), about 2.5:1(w:w), about 1:1(w:w), or about (0.8-1.8):1(w:w), about (0.9-1.7):1(w:w), or about (1-1.4):1; and / or albumin:SN-38 is about (5-25):1(w:w), about (5-20):1(w:w), about (6-19):1(w:w), about (7-18):1(w:w), about (8-16):1(w:w), about (9-14):1(w:w), or about (10-12):1(w:w), or about (9-21):1(w:w), about (9-20):1(w:w), about (11-18):1(w:w), or about (11.1-17.3):1(w:w); and / or albumin:cholesterol is about (5-25):1(w:w), about (6-20):1(w:w), about (7-18):1(w:w), about (8-16):1(w:w), about (9-14):1(w:w), about (10-12):1(w:w), or about (6-21):1(w:w), about (6.7-13):1(w:w), about (7-13):1(w:w), or about (11-12.7):1(w:w); and / or Span 20:SN-38 is about (5-40):100(w:w), about (6-30):100(w:w), about (7-25):100(w:w), about (8-20):100(w:w), about (9-15):100(w:w), about (10-12):100(w:w), or about (5-10):100(w:w), about (5-9):100(w:w), about (6-8.6):100(w:w), about (6-8):100(w:w), or about (6.5-7):100(w:w); and / or With respect to the total amount of SN-38, cholesterol, and albumin in the composition, an SN-38 content of about 3 w / w% to about 10 w / w%, about 3.5 w / w% to about 9.5 w / w%, about 4 w / w%, about 4.5 w / w%, about 5 w / w%, about 5.5 w / w%, about 6 w / w%, about 6.5 w / w%, about 7 w / w%, about 7.5 w / w%, about 8 w / w%, about 8.5 w / w%, about 9 w / w%, or about 4 w / w% to about 10 w / w%, about 4.5 w / w% to about 9.5 w / w%, about 5 w / w% to about 9 w / w%, or about 7.5 w / w% to about 8 w / w%; and / or Cholesterol content is about 4 w / w% to about 18 w / w%, about 4.5 w / w% to about 17.5 w / w%, about 5 w / w%, about 5.5 w / w%, about 6 w / w%, about 6.5 w / w%, about 7 w / w%, about 7.5 w / w%, about 8 w / w%, 8.5 w / w%, about 9 w / w%, about 9.5 w / w%, about 10 w / w%, about 10.5 w / w%, about 11 w / w%, about 11.5 w / w%, about 12 w / w%, about 12.5 w / w%, about 13 w / w%, about 13.5 w / w%, about 14 w / w%, about 14.5 w / w%, about 15 w / w%, about 15.5 w / w%, about 16 w / w%, about 16.5 w / w%, about 17 w / w%, or about 4 w / w% to about 12.5 w / w%, about 4.5 w / w% to about 12 w / w%, about 7 w / w% to about 10 w / w%, or about 7.5 w / w% to about 8 w / w%; and / or The composition according to embodiment 10, characterized in that the albumin content is about 78 w / w% to about 92 w / w%, about 79 w / w%, about 80 w / w%, about 81 w / w%, about 82 w / w%, about 83 w / w%, about 84 w / w%, about 85 w / w%, about 86 w / w%, about 87 w / w%, about 88 w / w%, about 89 w / w%, about 90 w / w%, about 91 w / w%, or about 78 w / w% to about 93 w / w%, about 79 w / w% to about 91.5 w / w%, about 80 w / w% to about 90 w / w%, about 82 w / w% to about 89 w / w%, about 84 w / w% to about 88 w / w%, or about 84.5 w / w% to about 87.5 w / w%.

[0185] Embodiment 15 the SN-38 present in the nanoparticles comprises at least about 6% to about 12% by weight, e.g., about 7% to about 11%, about 8% to about 10%, about 8.3%, or about 9% by weight of the total amount of SN-38, cholesterol, and albumin in the composition; and / or 15. The composition of any one of embodiments 12 to 14, wherein the SN-38 present in the nanoparticles accounts for about 95 w / w% to about 99 w / w%, e.g., about 96 w / w% to about 99 w / w%, about 97 w / w% to about 99 w / w%, about 98 w / w% to about 99 w / w%, or about 99 w / w% or more of the total amount of SN-38 in the composition.

[0186] Embodiment 16 16. The composition according to any one of embodiments 1 to 15, wherein the composition is in liquid, semi-solid or solid form.

[0187] Embodiment 17 The composition is in a solid form, preferably a powder form, more preferably a lyophilized powder; Preferably, the composition according to any one of embodiments 1 to 16, characterized in that SN-38 is present in the composition in amorphous and / or nanocrystalline form.

[0188] Embodiment 18 the composition does not contain an additional stabilizer; or Preferably, the composition according to any one of embodiments 1 to 17 further comprises an additional stabilizer, such as a lyophilization stabilizer, wherein the additional stabilizer is present in an amount such that when the composition is reconstituted to form an aqueous composition (including solutions and emulsions), the content of the additional stabilizer is at least about 2 w / v%, such as at least about 3 w / v%, for example at least about 5 w / v%, about 5 w / v% to about 30 w / v%, about 10 w / v% to about 25 w / v%, or about 15 w / v% to about 20 w / v%.

[0189] Embodiment 19 The composition further comprises an additional stabilizer, based on the total amount of the composition: The content of the additional stabilizer is about 60 w / w% to about 98 w / w%, for example, about 65 w / w% to about 97 w / w%, about 68 w / w% to about 96 w / w%, about 69 w / w% to about 95 w / w%, about 70 w / w% to about 94 w / w%, about 71 w / w% to about 93 w / w%, about 72 w / w% to about 92 w / w%, about 73 w / w%, about 74 w / w% , about 75 w / w%, about 76 w / w%, about 77 w / w%, about 78 w / w%, about 79 w / w%, about 80 w / w%, about 81 w / w%, about 82 w / w%, about 83 w / w%, about 84 w / w%, about 85 w / w%, about 86 w / w%, about 87 w / w%, about 88 w / w%, about 89 w / w%, about 90 w / w%, or about 91 w / w%; For example, the composition according to embodiment 17 or 18, wherein the content of the additional stabilizer is about 70 w / w% to about 96 w / w%, about 70 w / w% to about 90 w / w%, about 72 w / w% to about 89 w / w%, about 74 w / w% to about 88 w / w%, about 76 w / w% to about 87 w / w%, or about 80 w / w% to about 96 w / w%, about 80 w / w% to about 86 w / w%, about 81 w / w% to about 86 w / w%, about 82 w / w% to about 85 w / w%, about 83 w / w% to about 84 w / w%, or about 84 w / w% to about 95 w / w%.

[0190] Embodiment 20 20. The composition of embodiment 18 or 19, wherein the additional stabilizer is selected from albumin (e.g., human serum albumin, recombinant human albumin, bovine serum albumin, and skim milk powder), monosaccharides, disaccharides, polysaccharides, mannitol, and any combination thereof, preferably mannitol, lactose, maltose, trehalose, dextran, glucose, and sucrose, and any composition thereof, preferably sucrose.

[0191] Embodiment 21 21. The composition of any one of embodiments 17-20, wherein the nanoparticles have an average particle size of about 50-200 nm, e.g., about 90-150 nm, about 95-140 nm, about 100-130 nm, about 105-125 nm, or about 110-120 nm, when the composition is reconstituted to form an aqueous composition (including solutions and emulsions) at about 0.1 μg / mL to about 30.0 mg / mL.

[0192] Embodiment 22 the composition is an aqueous composition in liquid form, including solutions and emulsions; The composition according to any one of the preceding embodiments, in particular in liquid form, comprising SN-38 in the form of nanocrystals and / or vesicles.

[0193] Embodiment 23 With respect to the total amount of the composition, the content of SN-38 is about 0.1 μg / mL to about 30.0 mg / mL, about 0.2 μg / mL to about 27.0 mg / mL, about 0.5 μg / mL to about 24.0 mg / mL, about 1.0 μg / mL to about 21.0 mg / mL, about 5.0 μg / mL to about 18.0 mg / mL, about 10.0 μg / mL to about 15.0 mg / mL, about 20.0 μg / mL to about 12 mg / mL, about 25.0 μg / mL to about 9 mg / mL, about 50.0 μg / mL to about 6.0 mg / mL, or about 100.0 μg / mL to about 3.0 mg / mL; and / or a lipid content of about 0.05 μg / mL to about 100.0 mg / mL, about 0.1 μg / mL to about 90.0 mg / mL, about 0.25 μg / mL to about 80.0 mg / mL, about 0.5 μg / mL to about 70.0 mg / mL, about 2.5 μg / mL to about 60.0 mg / mL, about 5.0 μg / mL to about 50.0 mg / mL, about 10.0 μg / mL to about 40.0 mg / mL, about 12.5 μg / mL to about 30.0 mg / mL, about 25.0 μg / mL to about 20.0 mg / mL, or about 50.0 μg / mL to about 10.0 mg / mL; and / or 23. The composition according to embodiment 22, characterized in that the albumin content is about 3.0 μg / mL to about 300.0 mg / mL, about 6.0 μg / mL to about 270.0 mg / mL, about 15.0 μg / mL to about 240.0 mg / mL, about 30.0 μg / mL to about 210.0 mg / mL, about 150.0 μg / mL to about 180.0 mg / mL, about 300.0 μg / mL to about 150.0 mg / mL, about 600.0 μg / mL to about 120.0 mg / mL, about 750.0 μg / mL to about 90.0 mg / mL, about 1500.0 μg / mL to about 60.0 mg / mL, or about 3.0 mg / mL to about 30.0 mg / mL.

[0194] Embodiment 24 With respect to the total amount of the composition, the content of SN-38 is about 100.0 μg / mL to about 3.0 mg / mL, for example, about 200.0 μg / mL to about 2.5 mg / mL, about 300.0 μg / mL to about 2.0 mg / mL, about 400.0 μg / mL to about 1.5 mg / mL, about 500.0 μg / mL to about 1.0 mg / mL, or about 600 μg / mL to about 800 μg / mL; and / or the lipid content is about 50.0 μg / mL to about 10.0 mg / mL, for example, about 100.0 μg / mL to about 8.0 mg / mL, about 200.0 μg / mL to about 6.0 mg / mL, about 300.0 μg / mL to about 4.0 mg / mL, about 400.0 μg / mL to about 3.0 mg / mL, about 500.0 μg / mL to about 2.5 mg / mL, about 600.0 μg / mL to about 2.0 mg / mL, about 700.0 μg / mL to about 1.5 mg / mL, about 800 μg / mL to about 1.0 mg / mL, or about 200 μg / mL to about 1.5 mg / mL; and / or The composition according to embodiment 22 or 23, characterized in that the albumin content is about 3.0 mg / mL to about 30.0 mg / mL, for example, about 4.0 mg / mL to about 25.0 mg / mL, about 5.0 mg / mL to about 20.0 mg / mL, about 6.0 mg / mL to about 15.0 mg / mL, about 7.0 mg / mL to about 12.0 mg / mL, or about 8.0 mg / mL to about 10.0 mg / mL.

[0195] Embodiment 25 25. The composition of any one of embodiments 22 to 24, wherein the nanoparticles have an average particle size of about 50 to 200 nm, e.g., about 90 to 150 nm, about 95 to 140 nm, about 100 to 130 nm, about 105 to 125 nm, or about 110 to 120 nm.

[0196] Embodiment 26 26. The composition of any one of embodiments 22 to 25, wherein after storage at 4°C for 24 hours, the nanoparticles have an average particle size of about 50 to 200 nm, such as about 90 to 150 nm or about 100 to 130 nm.

[0197] Embodiment 27 27. The composition of any one of embodiments 22 to 26, wherein the nanoparticles have a particle size distribution index (PDI) of about 0.30 or less, such as about 0.2 or less, about 0.10 or less, or about 0.01 or less.

[0198] Embodiment 28 28. The composition according to any one of embodiments 22 to 27, wherein the composition has a zeta potential of about −35 mV to about −20 mV, such as about −31 mV.

[0199] Embodiment 29 29. The composition of any one of embodiments 22-28, wherein the nanoparticles do not disintegrate when the composition is diluted (e.g., with 1xPBS at a pH of about 7.4) to yield an SN-38 content of about 4 μg / mL or less, e.g., about 2 μg / mL or less, about 1 μg / mL or less, about 0.4 μg / mL or less, about 0.1 μg / mL or less, about 0.04 μg / mL or less, about 0.02 μg / mL or less, or about 0.01 μg / mL or less in the diluted composition.

[0200] Embodiment 30 the composition does not contain an additional stabilizer; or Preferably, the composition according to any one of embodiments 22 to 29 further comprises an additional stabilizer, and the content of the additional stabilizer relative to the total amount of the composition is at least about 2 w / v%, for example, at least about 3 w / v%, at least about 5 w / v%, about 5 w / v% to about 30 w / v%, about 10 w / v% to about 25 w / v%, or about 15 w / v% to about 20 w / v%.

[0201] Embodiment 31 The composition of embodiment 30, wherein the additional stabilizer is selected from albumin (e.g., human serum albumin, recombinant human albumin, bovine serum albumin, and skim milk powder), monosaccharides, disaccharides, polysaccharides, mannitol, and any combination thereof, preferably mannitol, lactose, maltose, trehalose, dextran, glucose, and sucrose, and any composition thereof, preferably sucrose.

[0202] Embodiment 32 the ring-opened SN-38 in the composition accounts for about 2 wt. % or less, e.g., about 1.8 wt. % or less, of the total amount of SN-38; and / or 32. The composition of any one of embodiments 1 to 31, wherein albumin multimers are absent or substantially absent from the composition; e.g., wherein the monomeric form of albumin in the composition accounts for at least about 95% w / w of the total amount of albumin, e.g., at least about 96%, at least about 98%, at least about 99%, at least about 99.2%, at least about 99.4%, or at least about 99.5%.

[0203] Embodiment 33 The albumin is selected from human serum albumin (HSA), recombinant human albumin (rHA), bovine serum albumin, and porcine serum albumin; for example, the albumin comprises the amino acid sequence set forth in SEQ ID NO: 1; The composition according to any one of embodiments 1 to 32, characterized in that the albumin is preferably selected from human serum albumin (HSA) and recombinant human albumin (rHA).

[0204] Embodiment 34 (1) dissolving SN-38, lipid, and Span 20 in an organic solvent to form an organic phase; and preparing an aqueous solution of albumin as an aqueous phase; (2) mixing an organic phase with an aqueous phase to form an emulsion, the emulsion comprising nanoparticles in which albumin encapsulates at least a portion of the SN-38 and optionally at least a portion of the lipid; and (3) Removing the organic solvent from the emulsion to obtain a product containing nanoparticles. 34. A method for preparing a composition according to any one of embodiments 1 to 33, comprising:

[0205] Embodiment 35 (1) SN-38, lipids, and Span 20 were mixed in DMSO and C 1~3 A first organic solvent selected from alcohols (including methanol, ethanol, and isopropanol, and any combination thereof, preferably ethanol (EtOH)), and a second organic solvent selected from CHCl3 and a mixture of CH2Cl2 and CHCl3, wherein the second organic solvent is DMSO or C 1~3 dissolving the albumin in a mixed organic solvent having a volume ratio of about 1:20 (v / v) to about 20:1 (v / v), for example, about 1:5 to about 5:1 (v / v), about 1:2 to about 4:1 (v / v), about 1:1 to about 4:1 (v / v), about 1.5:1 (v / v) to about 3:1 (v / v), or about 2:1 (v / v) to 7:3 (v / v) to form an organic phase; preparing an aqueous solution of albumin as an aqueous phase; (2) mixing an organic phase with an aqueous phase to prepare an emulsion, the emulsion comprising nanoparticles in which albumin encapsulates at least a portion of the SN-38 and optionally at least a portion of the lipid; (3) removing the organic solvent; and (4) Optionally, sterilizing the product obtained in step (3) by filtering it, preferably through a filter membrane of about 0.2 μm. Includes; 35. The method of embodiment 34, optionally, wherein the second organic solvent is CHCl3 or a mixture of CH2Cl2 and CHCl3, and optionally, the volume ratio of CH2Cl2 to CHCl3 in the mixture is about 2:5 to 1:1, preferably about 2:5.

[0206] Embodiment 36 36. The method according to embodiment 34 or 35, wherein in step (2), the ratio of organic phase to aqueous phase is from about 1:2 (v / v) to about 1:50 (v / v), for example, from about 1:5 (v / v) to about 1:20 (v / v), from about 1:7 (v / v) to about 1:15 (v / v), from about 1:10 (v / v) to about 1:12 (v / v), for example, from about 1:5 (v / v) to about 1:12 (v / v), from about 1:5 (v / v) to about 1:12 (v / v), about 1:6 (v / v), about 1:7 (v / v), or about 1:10 (v / v).

[0207] Embodiment 37 Step (2) (2-1) dispersing an organic phase in an aqueous phase under shear to obtain a crude emulsion; and (2-2) Homogenizing the coarse emulsion under high pressure to obtain a fine emulsion containing nanoparticles 37. The method according to any one of embodiments 34 to 36, comprising:

[0208] Embodiment 38 the aqueous phase does not contain additional stabilizers; or the aqueous phase already contains additional stabilizer; or The method further comprises adding an additional stabilizer in step (2); 38. The method of any one of embodiments 34 to 37, wherein the additional stabilizer is present in an amount such that the content of the additional stabilizer in the product obtained in step (3) or (4) is at least about 2 w / v%, e.g., at least about 3 w / v%, at least about 5 w / v%, about 5 w / v% to about 30 w / v%, about 10 w / v% to about 25 w / v%, or about 15 w / v% to about 20 w / v%.

[0209] Embodiment 39 39. The method of embodiment 38, wherein the additional stabilizer is selected from albumin (e.g., human serum albumin, recombinant human albumin, bovine serum albumin, and skim milk powder), monosaccharides, disaccharides, polysaccharides, mannitol, and any combination thereof, preferably mannitol, lactose, maltose, trehalose, dextran, glucose, and sucrose, and any composition thereof, preferably sucrose.

[0210] Embodiment 40 40. The method of any one of embodiments 34 to 39, wherein the mixed organic solvents in step (1) are added to the aqueous phase before mixing the organic phase and the aqueous phase in step (2).

[0211] Embodiment 41 41. The method according to embodiment 40, wherein the volume of the added mixed solvent is equal to or less than the volume of the organic phase; for example, the volume ratio of the added mixed solvent to the organic phase is from about 1:1 (v / v) to about 1:5 (v / v), for example from about 1:2 (v / v) to about 1:4 (v / v), or about 1:3 (v / v).

[0212] Embodiment 42 In the organic phase in step (1), the concentration of SN-38 is about 5-17 mg / mL, e.g., about 5.25-12 mg / mL, about 7-12 mg / mL, or about 10 mg / mL; and / or The lipid concentration is about 3 to 50 mg / mL, for example, about 5 to 45 mg / mL, or about 7.5 to 30 mg / mL, about 10 to 25 mg / mL, or about 15 to 20 mg / mL; and / or 42. The method according to any one of embodiments 34 to 41, wherein the concentration of albumin in the aqueous phase is about 5 to 15 mg / mL, such as about 6 to 12 mg / mL or about 6 to 10 mg / mL.

[0213] Embodiment 43 In the organic phase in step (1), the concentration of SN-38 is about 4-10 mg / mL, for example, about 6-8 mg / mL; and / or The lipid concentration is about 10-20 mg / mL, for example about 15 mg / mL; and / or The concentration of Span 20 is about 0.3 to 6 mg / mL, for example about 0.3 to 2 mg / mL or about 0.6 to 1 mg / mL; and / or 42. The method according to any one of embodiments 34 to 41, wherein the concentration of albumin in the aqueous phase is about 8 to 30 mg / mL, such as about 12 to 20 mg / mL or about 16 to 18 mg / mL.

[0214] EMBODIMENT 44 Step (4): Optionally, the method according to any one of embodiments 34 to 43, further comprising sterilizing the product obtained in step (3), preferably by filtering through a filter membrane of about 0.2 μm.

[0215] Embodiment 45 (5) drying the product obtained in step (3) or (4), preferably by spray drying or freeze drying, to obtain a composition in solid form, preferably a powder, more preferably a freeze-dried powder. further including; Preferably, the method according to any one of embodiments 34 to 44, characterized in that SN-38 is present in the composition in amorphous form.

[0216] Embodiment 46 The method of embodiment 45, wherein step (5) further comprises adding an additional stabilizer according to embodiment 39 to the product obtained in step (3) or (4) before drying, wherein the additional stabilizer is present in an amount such that when the solid form obtained in step (5) is reconstituted to form an aqueous composition (including solutions and emulsions), the content of the additional stabilizer is at least about 2 w / v%, for example, at least about 3 w / v%, at least about 5 w / v%, between about 5 w / v% and about 30 w / v%, between about 10 w / v% and about 25 w / v%, or between about 15 w / v% and about 20 w / v%.

[0217] Embodiment 47 A pharmaceutical composition comprising the composition of any one of embodiments 1 to 33, and optionally a pharmaceutically acceptable carrier.

[0218] Embodiment 48 A pharmaceutical composition comprising the composition of any one of embodiments 1-33 in a dry state, and optionally a pharmaceutically acceptable carrier.

[0219] Embodiment 49 49. The pharmaceutical composition according to embodiment 48, wherein the drying is freeze-drying or spray-drying, preferably freeze-drying.

[0220] Embodiment 50 50. The pharmaceutical composition according to embodiment 48 or 49, which is in solid form, preferably a lyophilized powder, and which is preferably used for parenteral administration, more preferably for administration by intravenous injection.

[0221] Embodiment 51 Use of the composition according to any one of embodiments 1 to 33 or the pharmaceutical composition according to any one of embodiments 47 to 50 in the manufacture of a medicament for treating an SN-38-sensitive tumor in a subject, preferably selected from colorectal cancer, small cell lung cancer, lymphatic cancer, breast cancer (preferably triple-negative breast cancer), esophageal cancer, gastric cancer, liver cancer, renal cancer, pancreatic cancer, uterine cancer, and ovarian cancer.

[0222] Embodiment 52 The composition of any one of embodiments 1 to 33 or the pharmaceutical composition of any one of embodiments 47 to 50 for use in treating an SN-38-sensitive tumor in a subject, preferably selected from colorectal cancer, small cell lung cancer, lymphatic cancer, breast cancer (preferably triple-negative breast cancer), esophageal cancer, gastric cancer, liver cancer, renal cancer, pancreatic cancer, uterine cancer, and ovarian cancer.

[0223] Embodiment 53 A method for treating an SN-38-sensitive tumor in a subject, preferably selected from colorectal cancer, small cell lung cancer, lymphatic cancer, breast cancer (preferably triple-negative breast cancer), esophageal cancer, gastric cancer, liver cancer, renal cancer, pancreatic cancer, uterine cancer, and ovarian cancer, comprising the step of administering a therapeutically effective amount of the composition of any one of embodiments 1 to 33 or the pharmaceutical composition of any one of embodiments 47 to 50 to the subject.

[0224] EMBODIMENT 54 A kit comprising the composition according to any one of embodiments 1 to 33 or the pharmaceutical composition according to any one of embodiments 47 to 50.

[0225] Embodiment 55 the composition comprises SN-38, a lipid, and albumin, wherein the albumin encapsulates at least a portion of the SN-38 and optionally at least a portion of the lipid to form nanoparticles; The method is characterized in that Span 20 is added during the preparation of the composition; Optionally, the composition does not contain additional stabilizers; and / or Optionally, the improved properties include improved stability; for example, when the composition is in liquid form, the improved stability includes a reduction in the formation or content of albumin multimers (e.g., no or substantially no albumin multimers are present in the composition, or albumin multimers account for at most 5 w / w%, e.g., at most about 4%, at most about 2%, at most about 1.5%, at most about 1.2%, at most about 1.1%, at most about 1%, or at most about 0.8% of the total amount of albumin), and / or a reduction in particle size of the nanoparticles during preparation, storage, and / or use of the composition; and / or Optionally, the composition is as defined in any one of embodiments 1 to 33. Methods for preparing compositions with improved properties.

[0226] Embodiment 56 (1) dissolving SN-38, lipid, and Span 20 in an organic solvent to form an organic phase; and preparing an aqueous solution of albumin as an aqueous phase; (2) mixing an organic phase with an aqueous phase to form an emulsion, the emulsion comprising nanoparticles in which albumin encapsulates at least a portion of the SN-38 and optionally at least a portion of the lipid; and (3) Removing the organic solvent from the emulsion to obtain a product containing nanoparticles. 56. The method of embodiment 55, comprising:

[0227] Embodiment 57 (1) SN-38, lipids, and Span 20 were mixed in DMSO and C 1~3 A first organic solvent selected from alcohols (including methanol, ethanol, and isopropanol, and any combination thereof, preferably ethanol (EtOH)), and a second organic solvent selected from CHCl3 and a mixture of CH2Cl2 and CHCl3, wherein the second organic solvent is DMSO or C 1~3 dissolving the albumin in a mixed organic solvent having a volume ratio of about 1:20 (v / v) to about 20:1 (v / v), for example, about 1:5 to about 5:1 (v / v), about 1:2 to about 4:1 (v / v), about 1:1 to about 4:1 (v / v), about 1.5:1 (v / v) to about 3:1 (v / v), or about 2:1 (v / v) to 7:3 (v / v) to form an organic phase; preparing an aqueous solution of albumin as an aqueous phase; (2) mixing an organic phase with an aqueous phase to prepare an emulsion, the emulsion comprising nanoparticles in which albumin encapsulates at least a portion of the SN-38 and optionally at least a portion of the lipid; (3) removing the organic solvent; and (4) Optionally, sterilizing the product obtained in step (3) by filtering it, preferably through a filter membrane of about 0.2 μm. Includes; 57. The method of embodiment 56, optionally wherein the second organic solvent is CHCl3 or a mixture of CH2Cl2 and CHCl3, and optionally wherein the volume ratio of CH2Cl2 to CHCl3 in the mixture is about 2:5 to 1:1, preferably about 2:5.

[0228] The inventors have discovered that the present invention reduces the number of high-pressure homogenization steps during preparation, particularly in scaled-up processes, such as pilot-scale preparations, and substantially reduces the particle size of nanoparticles in the composition, increases filtration flux, stabilizes the particle size of nanoparticles after disintegration, and reduces raw material loss and costs. Furthermore, the present invention further controls the albumin content in the composition, thereby controlling the particle size of nanoparticles. Therefore, the particle size of nanoparticles in the composition of the present invention approaches a particle size suitable for drug preparation. Furthermore, since the presence of Span 20 prevents the formation of albumin multimers during storage, the composition of the present invention has low immunogenicity, high safety, and excellent storage stability. [Example]

[0229] The present invention is further described in the following examples, which are used only to illustrate the present invention and are not intended to limit the present invention in any way.

[0230] The abbreviations used in the examples have the following meanings:

[0231] [Table 1]

[0232] Methods for measuring various parameters of the products prepared in the examples 1. Measurement of particle size and particle size distribution The particle size and size distribution of nanoparticles in the samples were measured using a Malvern Nano ZSE particle size potentiometer. The laser light emitted by this instrument had a wavelength of 633 nm, and the included angle between the incident light and the scattered light was 173°. The parameters were set as follows: protein as sample material; water as dispersant; measurement temperature 25°C; and automatic scanning detection. Each sample was measured three times in parallel, and the results were averaged.

[0233] 2. Zeta potential measurement The zeta potential of nanoparticles in the samples was measured using a Malvern Nano ZSE particle size potentiometer. The parameters were set as follows: protein as sample material; and water as dispersant. The DTS1070 sample pool was selected; the measurement temperature was set to 25°C; and automatic scanning detection was employed. Before detection, the test samples were diluted 10 times by volume with deionized water. Each sample was measured three times in parallel, and the results were averaged. 3. Measurement of SN-38 content in the system 10 mg of the prepared sample was diluted 5-fold with deionized water, then 10-fold with isopropanol, and extracted by ultrasound for 15 minutes, followed by centrifugation at 10,000 rpm for 12 minutes. The supernatant was collected for HPLC analysis to determine the SN-38 content in the system. The peak area was fitted to a standard curve to calculate the SN-38 content in the system.

[0234] The HPLC conditions were as shown in Table 1.

[0235] [Table 2]

[0236] The gradient elution conditions were as shown in Table 2.

[0237] [Table 3]

[0238] A typical chromatogram obtained by measuring the content of SN-38 is shown in FIG. 1 (taking Example 1 as an example).

[0239] 4. Determination of albumin content in the system The albumin content in the system was measured using the BCA method. BSA was used as the standard substance, and the samples were diluted 10-fold. 25 μL of the diluted sample was taken, 200 μL of detection solution was added, and the mixture was mixed homogeneously by shaking on a shaker. The microwell plate was then sealed and incubated at 37°C for 120 minutes. The absorbance was measured at 562 nm on a microplate reader, and the albumin concentration in the sample was calculated according to the standard curve.

[0240] 5. Determination of cholesterol content in the system The cholesterol content in the sample was measured by HPLC. The method for diluting the sample was the same as that for measuring the SN-38 content above. The chromatographic conditions were as shown in Table 3.

[0241] [Table 4]

[0242] A typical chromatogram obtained by measuring the cholesterol content is shown in FIG. 2 (taking Example 1 as an example).

[0243] 6. Calculation of Drug Loading Capacity (LD) and Encapsulation Efficiency (EE)

number

[0244] In the formula, the total SN-38 content in the system is measured by the method described in "3. Measurement of SN-38 content in the system."

[0245] The content of free SN-38 in the system was measured by HPLC after extraction by solid phase extraction. The specific method of solid phase extraction was as follows: 1) SPE plug (Select Core™ HLB, 1 mL) activation: Activated first with 3 mL of methanol, then with 3 mL. 2) 200 μL of the sample to be separated was loaded and passed through the column under gravity. 3) Elution was carried out under gravity with 2 mL of water. 4) Elution was carried out under gravity with 2 mL of methanol. The methanol phase was collected to obtain the detection solution of free SN-38.

[0246] 7. Discovery of two structures of SN-38 The two structures of SN-38, which differ in their activity, are shown schematically below: [ka]

[0247] The method for treating the samples was the same as that for the above measurement of SN-38 content, and the HPLC chromatographic conditions were as shown in Table 4.

[0248] [Table 5]

[0249] The gradient elution conditions were as shown in Table 5.

[0250] [Table 6]

[0251] A typical chromatogram obtained by measuring the content of SN-38 with various structures is shown in FIG. 3 (taking Example 1 as an example).

[0252] The proportion of ring-opened SN-38 in a sample can be calculated according to the peak area ratio of the ring-open structure to the lactone ring structure in the chromatogram. The proportion of ring-opened SN-38 in the samples prepared in the Examples according to the present invention was all less than 2.0%.

[0253] 8. Measuring Albumin Aggregates in Samples SEC-HPLC was used to measure the aggregation of albumin in the samples. 5 μl of the prepared sample was directly taken for detection, and the chromatographic conditions were as shown in Table 6.

[0254] [Table 7]

[0255] A typical chromatogram obtained by measuring albumin aggregates is shown in Figure 4 (taking Example 1 as an example). The results showed that there were no albumin multimers in the sample, and only a small amount of dimers, indicating that this product did not cause immunogenicity due to albumin multimers.

[0256] 9. Experiments on stability and collapse The stability of the samples was mainly measured by storing the prepared samples separately at room temperature and 4°C and observing the samples intermittently to see if there was any obvious precipitation or precipitate in the samples. At the same time, samples were taken for particle size and particle size distribution detection to investigate the size change of nanoparticles in the samples.

[0257] Disintegration experiments were conducted to investigate the binding stability of albumin and SN-38 in the samples. The samples were diluted with 1x PBS at pH 7.4, and the particle size and particle size distribution of the samples at various dilution ratios were measured to investigate at what dilution ratio the nanoparticles would disintegrate and the SN-38 raw material would precipitate. The greater the dilution ratio, the better the stability of the nanoparticles.

[0258] 10.XRD detection method The crystalline form of the active pharmaceutical ingredient (API) in the samples was evaluated using X-ray diffraction (Bruker, D8 ADVANCE). The lyophilized albumin powder, lyophilized albumin-SN-38 powder, and SN-38 crystalline forms were detected separately. Cu-Kα radiation was used to scan the 2θ range from 2° to 40° at a rate of 2° / min.

[0259] 11. Electron Microscopy Detection Sample preparation conditions: The API concentration of each sample was adjusted to 2 mg / mL with water for injection. The temperature was 4°C, the humidity was 100%, the blotting time was 9 seconds, and the blotting force was 3. The morphology of vesicles and crystals in the samples was observed under 120 Kv using a cryogenic transmission electron microscope (Talos L120C).

[0260] According to the specific situation, the above measurement methods were selected to measure the samples prepared in the following examples.

[0261] Test materials: Unless otherwise specified, SN-38 used in the following examples was provided by Sichuanxieli Pharmaceutical Co., Ltd.; cholesterol was provided by Jiangsu Southeast Nanomaterials Co., Ltd.; rHA was provided by North China Pharmaceutical Company Ltd.; HSA was provided by Guang Dong Shuang Lin Bio-Pharmacy Co., Ltd.; and irinotecan hydrochloride injection (CPT-11, 60 mg / kg) was provided by Jiangsu Hengrui Pharmaceuticals Co., Ltd.

[0262] Unless otherwise stated, in the following animal experimental studies, the doses of rHA-SN-38 or HSA-SN-38 products and CPT-11 were based on the active ingredient; the solvent was water for injection, which was used as the blank control.

[0263] Example 1: Preparation of rHA-SN-38 Product 1 1. Preparation Process 1) Prepare an organic solvent of EtOH / CHCl3 with a volume ratio of 2 / 3; 2) Take 21 mg of SN-38 and 30 mg of cholesterol, add 3 mL of the organic solvent from step 1), and dissolve completely to obtain a drug solution; 3) A total volume of approximately 21 mL of an rHA aqueous solution was prepared using deionized water as the aqueous phase so that the total content of rHA in the aqueous phase was 200 mg; 4) Shear dispersion: After mixing the drug solution from step 2) with the aqueous phase from step 3), shear dispersion is carried out for 10-15 minutes to obtain a crude emulsion; 5) The crude emulsion is transferred to a high-pressure homogenizer and homogenized under a pressure of 1300-1500 bar for 2-7 minutes; 6) Rotary evaporation at 40°C to 45°C for 4 to 8 minutes; 7) Filtration was performed through a 0.2 μm PES syringe filter membrane (Sartorius Pharm.) Parameters such as particle size, encapsulation efficiency, and drug loading of product samples were measured before and after filtration.

[0264] 2.Measurement results The measurement results of the sample prepared in Example 1 are shown in Table 7.

[0265] [Table 8]

[0266] Example 2: Preparation of lyophilized formulation of HSA-SN-38 Product 1 and its reconstitution solution 1. Preparation Process 1) Prepare an organic solvent of EtOH / CHCl3 with a volume ratio of 2 / 3; 2) Take 21 mg of SN-38 and 30 mg of cholesterol, add 3 mL of the organic solvent from step 1), and dissolve completely to obtain a drug solution; 3) A total volume of about 32 mL of an aqueous HSA solution was prepared using deionized water as the aqueous phase, so that the total content of HSA in the aqueous phase was 200 mg; 4) Shear dispersion: After mixing the drug solution from step 2) with the aqueous phase from step 3), shear dispersion is carried out for 10-15 minutes to obtain a crude emulsion; 5) The crude emulsion is transferred to a high-pressure homogenizer and homogenized under a pressure of 1300-1500 bar for 2-7 minutes; 6) Rotary evaporation at 40°C to 45°C for 4 to 8 minutes; 7) Add sucrose to the product obtained in step 6) and stir to completely dissolve the sucrose to obtain a sucrose concentration of 30 mg / mL; 8) The solution was filtered through a 0.2 μm PES syringe filter membrane, filled into vials, and freeze-dried under reduced pressure to obtain a freeze-dried formulation of HSA-SN-38 nanoparticles. The freeze-dried product was subjected to XRD analysis. Two lyophilized samples were taken, one of which was diluted with deionized water to the same concentration of SN-38 as before lyophilization (reconstituted solution 1), and the other was diluted with deionized water to the same concentration of SN-38 as before lyophilization (reconstituted solution 2). Parameters such as particle size and encapsulation efficiency of the two reconstituted solutions were measured.

[0267] 2.Measurement results (1) Measurement results of parameters of the reconstituted solution The results of measuring the parameters of the reconstituted solution of the lyophilized product prepared in Example 2 are shown in Table 8.

[0268] [Table 9]

[0269] The results show that when the concentration of the active ingredient is increased by up to 6 times after reconstitution, the particle size only increases slightly and remains below 200 nm. Other properties of the sample are not significantly affected. Therefore, the freeze-dried formulation of the present application can be diluted to various concentrations according to the needs of use.

[0270] (2)XRD analysis results The freeze-dried product, SN-38 crystals, and HSA were subjected to XRD detection, and the results are shown in Figure 5.

[0271] The results, based on a comparison of the measurement results for HSA-SN-38 and SN-38, indicate that the SN-38 in the lyophilized powder was in an amorphous state. It has been reported that amorphous active pharmaceutical ingredients are more soluble and have higher bioavailability than crystalline forms (e.g., Wang, D., Liang, N., Kawashima, Y. et al. Biotin-modified bovine serum albumin nanoparticles as a potential drug delivery system for paclitaxel. J Mater Sci 54, 8613-8626 (2019)). The formulation of the present application exhibits the advantage of a faster dissolution rate and higher bioavailability of SN-38 compared to crystalline forms of SN-38.

[0272] Example 3: Preparation of rHA-SN-38 Product 2 1. Preparation Process 1) Prepare an organic solvent of EtOH / CHCl3 with a volume ratio of 2 / 3; 2) Take 42 mg of SN-38 and 60 mg of cholesterol, add 3 mL of the organic solvent from step 1), and dissolve completely to obtain a drug solution; 3) A total volume of approximately 21 mL of an rHA aqueous solution was prepared using deionized water as the aqueous phase so that the total content of rHA in the aqueous phase was 500 mg; 4) Shear dispersion: After mixing the drug solution from step 2) with the aqueous phase from step 3), shear dispersion is carried out for 10-15 minutes to obtain a crude emulsion; 5) The crude emulsion is transferred to a high-pressure homogenizer and homogenized under a pressure of 1300-1500 bar for 2-7 minutes; 6) Rotary evaporation at 40°C to 45°C for 4 to 8 minutes; 7) Filtration was performed through a 0.2 μm PES syringe filter membrane. Before and after filtration, the product samples were measured for parameters such as particle size, encapsulation efficiency, and drug loading.

[0273] 2.Measurement results The measurement results for Product 2 are shown in Table 9.

[0274] [Table 10]

[0275] Example 4: Preparation of rHA-SN-38 Product 3 1. Preparation Process 1) Prepare an organic solvent of EtOH / CHCl3 with a volume ratio of 2 / 3; 2) Take 42 mg of SN-38 and 60 mg of cholesterol, add 3 mL of the organic solvent from step 1), and dissolve completely to obtain a drug solution; 3) A total volume of approximately 21 mL of an rHA aqueous solution was prepared using deionized water as the aqueous phase so that the total content of rHA in the aqueous phase was 200 mg; 4) Shear dispersion: After mixing the drug solution from step 2) with the aqueous phase from step 3), shear dispersion is carried out for 10-15 minutes to obtain a crude emulsion; 5) The crude emulsion is transferred to a high-pressure homogenizer and homogenized under a pressure of 1300-1500 bar for 2-7 minutes; 6) Rotary evaporation at 40°C to 45°C for 4 to 8 minutes; 7) Filtration was performed through a 0.2 μm PES syringe filter membrane. Before and after filtration, the product samples were measured for parameters such as particle size, encapsulation efficiency, and drug loading.

[0276] 2.Measurement results The measurement results for Product 3 are shown in Table 10.

[0277] [Table 11]

[0278] Example 5: Preparation of rHA-SN-38 Product 4 1. Preparation Process 1) Prepare an organic solvent of EtOH / CHCl3 with a volume ratio of 2 / 3; 2) Take 10 mg of SN-38 and 60 mg of cholesterol, add 3 mL of the organic solvent from step 1), and dissolve completely to obtain a drug solution; 3) A total volume of approximately 21 mL of an rHA aqueous solution was prepared using deionized water as the aqueous phase so that the total content of rHA in the aqueous phase was 150 mg; 4) Shear dispersion: After mixing the drug solution from step 2) with the aqueous phase from step 3), shear dispersion is carried out for 10-15 minutes to obtain a crude emulsion; 5) The crude emulsion is transferred to a high-pressure homogenizer and homogenized under a pressure of 1300-1500 bar for 2-7 minutes; 6) Rotary evaporation at 40°C to 45°C for 4 to 8 minutes; 7) Filtration was performed through a 0.2 μm PES syringe filter membrane. Before and after filtration, the product samples were measured for parameters such as particle size, encapsulation efficiency, and drug loading.

[0279] 2.Measurement results The measurement results for Product 4 are shown in Table 11.

[0280] [Table 12]

[0281] Example 6: Preparation of rHA-SN-38 product in the absence of lipids The rHA-SN-38 product of Example 6 was prepared and tested according to the preparation process of Example 1, except that no lipid (e.g., cholesterol) was added in step 2) of the preparation process of Example 1. The measurement results of the parameters of the rHA-SN-38 product are shown in Table 12.

[0282] [Table 13]

[0283] The results show that the sample prepared without adding lipid had poor stability, with the particle size of the sample increasing by 69.12% after 24 hours of storage in a refrigerator at 4°C. In contrast, the particle size of the sample prepared in Example 1 (with added lipid) increased by only 10.91% after 24 hours of storage in a refrigerator at 4°C. This indicates that the addition of cholesterol significantly improved the stability of the sample. Furthermore, the drug loading and encapsulation efficiency without adding lipid were lower than those with added cholesterol.

[0284] Example 7: Preparation of rHA-SN-38 products with the addition of other lipids Two rHA-SN-38 products were prepared and tested according to the preparation process of Example 1, except that cholic acid or palmitic acid was used instead of cholesterol in step 2) of the preparation process of Example 1. The measurement results of the parameters of the rHA-SN-38 products are shown in Table 13.

[0285] [Table 14]

[0286] The results show that the samples prepared by the same preparation process using cholic acid or palmitic acid as lipids had larger particle sizes, and the sample with palmitic acid added had larger particle sizes and a wider particle size distribution than the sample with cholic acid added. After processing through the membrane, the content of nanoparticles in the sample was very low and the particle size distribution was very large, so the desired results regarding particle size and PDI could not be obtained.

[0287] Example 8: Investigation of various organic solvent systems An rHA-SN-38 product was prepared and tested according to the preparation process of Example 1, except that the organic solvent system shown in Table 14 was used instead of the organic solvent in step 1) of the preparation process of Example 1. The particle size of the resulting product was also measured after overnight storage in a refrigerator at 4°C. The measurement results for the parameters of the resulting rHA-SN-38 product are shown in Table 14.

[0288] [Table 15]

[0289] The results show that the sample prepared using chloroform / ethanol as the organic solvent had the smallest particle size and the highest drug loading.

[0290] Example 9: Investigation of various ratios of organic solvents The rHA-SN-38 products were prepared and tested according to the preparation process of Example 1, except that various ratios of EtOH / CHCl3 shown in Table 15 were used instead of the organic solvent in step 1) of the preparation process of Example 1, and the amount of rHA used in the system was adjusted to 300 mg. The measurement results of the parameters of the obtained rHA-SN-38 products are shown in Table 15.

[0291] [Table 16]

[0292] It can be seen that rHA-SN-38 products with sufficient particle size, drug loading, and encapsulation efficiency can be obtained with various ratios of EtOH / CHCl3 organic solutions.

[0293] Example 10: Investigation of various cholesterol concentrations An rHA-SN-38 product was prepared and tested according to the preparation process of Example 1, except that the amount of cholesterol used in step 2) of the preparation process of Example 1 was adjusted to the amount shown in Table 16. The measurement results of the parameters of the obtained rHA-SN-38 product are shown in Table 16.

[0294] [Table 17]

[0295] The results show that when the amount of cholesterol used in the formulation was increased, the particle size of the sample after passing through the membrane became smaller, the encapsulation efficiency of SN-38 in the sample became higher, and the drug loading amount became higher.

[0296] Example 11: Investigation of various concentrations of SN-38 An rHA-SN-38 product was prepared and tested according to the preparation process of Example 1, except that the amount of SN-38 used in step 2) of the preparation process of Example 1 was adjusted to the amount shown in Table 17. The measurement results of the parameters of the obtained rHA-SN-38 product are shown in Table 17.

[0297] [Table 18]

[0298] The results show that increasing the concentration of SN-38 in the formulation had little effect on particle size, but increased drug loading.

[0299] Example 12: Investigation of different rHA concentrations in the aqueous phase An rHA-SN-38 product was prepared and tested according to the preparation process of Example 1, except that the amount of rHA or the volume of the aqueous phase in step 3) of the preparation process of Example 1 was adjusted to the values shown in Table 18. The measurement results of the parameters of the obtained rHA-SN-38 product are shown in Table 18.

[0300] [Table 19]

[0301] The results show that increasing the amount of albumin slightly decreased the drug loading, and the encapsulation efficiency was little affected.

[0302] Example 13: Investigation of various volume ratios of organic solvent to aqueous phase An rHA-SN-38 product was prepared and tested according to the preparation process of Example 1, except that the volume of the organic solvent in step 2) or the volume of the aqueous phase in step 3) of the preparation process of Example 1 was adjusted to the value shown in Table 19. The measurement results of the parameters of the obtained rHA-SN-38 product are shown in Table 19.

[0303] [Table 20]

[0304] The results show that the smaller the volume ratio of organic solvent:aqueous phase, the higher the drug loading of the product and the higher the encapsulation efficiency of SN-38.

[0305] Example 14: Preparation of rHA-SN-38 product containing additional stabilizers 1. Preparation Process 1) Prepare an organic solvent of EtOH / CHCl3 with a volume ratio of 2 / 3; 2) Take 21 mg of SN-38 and 30 mg of cholesterol, add 3 mL of the organic solvent from step 1), and dissolve completely to obtain a drug solution; 3) Prepare an HSA solution with deionized water and add sucrose or glucose as a stabilizer to form an aqueous phase (approximately 32 mL) with a total HSA content of 200 mg and a sucrose or glucose concentration in the final product as shown in Table 20; 4) Shear dispersion: After mixing the drug solution from step 2) with the aqueous phase from step 3), shear dispersion is carried out for 10-15 minutes to obtain a crude emulsion; 5) The crude emulsion is transferred to a high-pressure homogenizer and homogenized under a pressure of 1300-1500 bar for 2-7 minutes; 6) Rotary evaporation at 40°C to 45°C for 4 to 8 minutes; 7) Filtration was performed through a 0.2 μm PES syringe filter membrane. Before and after filtration, the product samples were measured for parameters such as particle size, encapsulation efficiency, and drug loading. 8) The particle size of the resulting product was measured again after overnight storage in a refrigerator at 4°C.

[0306] 2.Measurement results The measurement results of the parameters of the prepared HSA-SN-38 product are shown in Table 20.

[0307] [Table 21]

[0308] The results show that the products prepared by adding sucrose to the aqueous phase had relatively smaller particle sizes and better stability than those prepared with glucose, but lower drug loading than those prepared with glucose. For the same stabilizer, increasing its concentration further improved these parameters.

[0309] Furthermore, the inventors measured the effect of cyclodextrin (5%, 10%, and 15%) as a stabilizer and found that the prepared products were either cloudy due to precipitation or had particle sizes greater than 250 nm after 24 hours at room temperature. Therefore, cyclodextrin may not be suitable for use as a stabilizer.

[0310] Example 15: Preparation of HSA-SN-38 product by different preparation processes 1. Preparation Process 1) Prepare an organic solvent of EtOH / CHCl3 with a volume ratio of 2 / 3; 2) Take 21 mg of SN-38 and 30 mg of cholesterol, add 3 mL of the organic solvent from step 1), and dissolve completely to obtain a drug solution; 3) HSA solution was prepared by adding sucrose to deionized water to form an aqueous phase (approximately 32 mL) with a total content of 200 mg of HSA in the aqueous phase and a sucrose concentration of 10% (w / v) in the final product; 4) Shear dispersion: 1 mL of the organic solvent from step 1) was added to the aqueous phase from step 3) and dispersed under shear for 5 minutes, and then the drug solution from step 2) was added and continuously dispersed under shear for 5 minutes to obtain a crude emulsion; 5) The crude emulsion is transferred to a high-pressure homogenizer and homogenized under a pressure of 1300-1500 bar for 2-7 minutes; 6) Rotary evaporation at 40°C to 45°C for 4 to 8 minutes; 7) Filtration was performed through a 0.2 μm PES syringe filter membrane. Before and after filtration, the product samples were measured for parameters such as particle size, encapsulation efficiency, and drug loading. 8) The particle size of the resulting product was measured again after overnight storage in a refrigerator at 4°C.

[0311] 2.Measurement results The measurement results of the parameters of the prepared HSA-SN-38 product are shown in Table 21.

[0312] [Table 22]

[0313] The results show that the HSA-SN-38 product, which was prepared by first treating the albumin solution with a small amount of organic solvent and then adding the drug solution, exhibited superior parameters, especially increased drug loading.

[0314] Example 16: Preparation of rHA-SN-38 products prepared using other lipids An rHA-SN-38 product was prepared and tested according to the preparation process of Example 1, except that the lipids in Table 22 below (cholesteryl palmitate (Chol-PA, TCI Co., Ltd.); glyceryl monostearate (SA-Gly, damas-beta Co., Ltd.); and vitamin D3 (Aladdin Reagent Co., Ltd.)) were used instead of cholesterol in step 2) of the preparation process of Example 1. The particle size of the resulting product was also measured after overnight storage in a refrigerator at 4°C. The measurement results of the parameters of the rHA-SN-38 product are shown in Table 22.

[0315] [Table 23]

[0316] The results show that Chol-PA and SA-Gly can produce products with uniform particle size and high drug loading. The higher the amount of Chol-PA, the smaller the product particle size, the higher the absolute SN-38 recovery, and the higher the drug loading. However, the particle size distribution became broader. Products prepared with the addition of SA-Gly produced larger particle sizes, higher absolute SN-38 recovery, and higher drug loading, but were less stable.

[0317] Example 17: Preparation of rHA-SN-38 products prepared using lipid combinations An rHA-SN-38 product was prepared and tested according to the preparation process of Example 1, except that the lipid combinations shown in Table 23 below were used in place of cholesterol in step 2) of the preparation process of Example 1. The particle size of the resulting product was also measured after overnight storage in a refrigerator at 4°C. The measured parameters of the rHA-SN-38 product are shown in Table 23.

[0318] [Table 24]

[0319] The results show that adding Chol-PA to the formulation can reduce the particle size of the product, and the higher the addition amount, the smaller the particle size of the product and the better the stability. Products prepared by adding SA-Gly to the formulation had larger particle sizes but a narrower particle size distribution. Products prepared by adding both Chol-PA and SA-Gly had a narrower particle size distribution and good storage stability.

[0320] Example 18: Preparation of rHA-SN-38 products prepared with increasing proportions of lipids Except for the amount of cholesterol, which was 60 mg, the other raw materials in this example were the same as those in Example 1. To obtain the liquid and lyophilized powder formulations, the raw materials were divided into two groups: one group for preparing the liquid formulation by the method in Example 1, and the other group for preparing the lyophilized powder formulation by the method in Example 2 (without step 7). The measurement results of the parameters of the obtained liquid and lyophilized powder formulations of rHA-SN-38 are shown in the table below:

[0321] [Table 25]

[0322] Example 19: Large-scale preparation of HSA-SN-38 product A pilot-scale preparation process was investigated based on the small-scale preparation described above, and the HSA-SN-38 product of this example was prepared under large-scale conditions.

[0323] 1. Preparation Process 1) Prepare a mixed organic solvent of EtOH (152 mL) and CHCl3 (228 mL); 2) Take 3.36 g of SN-38 and 4.8 g of cholesterol, completely dissolve them in the organic solvent of step 1), and incubate them at 50°C for more than 30 minutes to obtain a drug solution; 3) Prepare an aqueous solution (3360 mL) of HSA (32 g) using deionized water as the aqueous phase; 4) Shear dispersion: After mixing the drug solution from step 2) with the aqueous phase from step 3), shear dispersion is carried out for 10-15 minutes to obtain a crude emulsion; 5) The crude emulsion was transferred to a high-pressure homogenizer and homogenized under a pressure of 900-1200 bar for 10 cycles; 6) Adding sucrose solution (100g / L, 2.16L) to the product obtained in step 5) and mixing; 7) The mixture obtained in step 6) was evaporated by rotary evaporation under 60-70 mbar, and the sample was concentrated using tangential flow ultrafiltration (Suzhou Saiensi Instrument Co., Ltd.); 8) A bag filter (Sartorius, SARTOBRAN P) was used for filtration. Samples were taken before and after filtration to measure particle size and encapsulation efficiency. The filtrate was filled into vials and freeze-dried under reduced pressure to obtain a freeze-dried formulation of HSA-SN-38 nanoparticles.

[0324] 2.Measurement results (1) Particle size results The particle size of the filtrate sample prepared in step 8) was 124.6 nm (PDI=0.187). (2) Encapsulation effect

[0325] [Table 26]

[0326] 3. Collapse Experiment The prepared lyophilized formulation was reconstituted with 1x PBS at pH 7.4 to an SN-38 concentration of 1 mg / mL, and then diluted to 100 μg / mL, 10 μg / mL, 1 μg / mL, 0.1 μg / mL, and 0.01 μg / mL, and the particle size and particle size distribution at various concentrations were measured.

[0327] The results are shown in Figure 6. The results show that the HSA-SN-38 nanoparticles began to disintegrate when the SN-38 concentration was less than 1 µg / mL and slowly disintegrated when the SN-38 concentration reached 0.01 µg / mL. This indicates that the lyophilized HSA-SN-38 product prepared in Example 19 exhibited excellent stability, which was significantly superior to other albumin nanoparticles currently on the market.

[0328] Example 20: Large-scale preparation of HSA-SN-38 product A pilot-scale preparation process was investigated based on the small-scale preparation described above, and the HSA-SN-38 product of this example was prepared under large-scale conditions.

[0329] 1. Preparation Process 1) Prepare a mixed organic solvent of EtOH (222 mL) and CHCl3 (333 mL); 2) Take 4.41 g of SN-38 and 6.30 g of cholesterol, dissolve them completely in 480 mL of the organic solvent from step 1), incubate them at 50°C for more than 30 minutes, and then mix them with the remaining organic solvent to obtain a drug solution; 3) Prepare an aqueous solution (3360 mL) of HSA (32 g) using deionized water as the aqueous phase; 4) Shear dispersion: After mixing the drug solution from step 2) with the aqueous phase from step 3), shear dispersion is carried out for 10-15 minutes to obtain a crude emulsion; 5) The crude emulsion was transferred to a high-pressure homogenizer and homogenized under a pressure of 900-1200 bar for 10 cycles; 6) adding sucrose solution (36 g / L, 6 L) to the product obtained in step 5) and mixing; 7) The mixture obtained in step 6) was evaporated by rotary evaporation under 60-70 mbar, and the sample was concentrated using tangential flow ultrafiltration (Suzhou Saiensi Instrument Co., Ltd.); 8) A bag filter (Sartorius, SARTOBRAN P) was used for filtration. Samples were taken before and after filtration to measure particle size and encapsulation efficiency. The filtrate was filled into vials and freeze-dried under reduced pressure to obtain a freeze-dried formulation of HSA-SN-38 nanoparticles.

[0330] 2.Measurement results (1) Particle size results The particle size of the filtrate sample prepared in step 8) was 146.9 nm (PDI=0.208). (2) Encapsulation effect

[0331] [Table 27]

[0332] 3. Collapse Experiment The prepared lyophilized formulation was reconstituted with water for injection to an SN-38 concentration of 1 mg / mL, and then gradient diluted to 100 μg / mL, 10 μg / mL, 1 μg / mL, 0.1 μg / mL, and 0.01 μg / mL, and the particle size and particle size distribution at various concentrations were measured. The results are shown in Figure 7. The results show that, similar to the product of Example 19, the HSA-SN-38 product of this example began to disintegrate when the SN-38 concentration was below about 1 µg / mL, with a significant broadening of the particle size distribution and rapid disintegration when the SN-38 concentration reached 0.1 µg / mL. The HSA-SN-38 product of this example still exhibited good stability, clearly superior to other commercially available albumin nanoformulations.

[0333] Example 21: Toxicity study of the rHA-SN-38 product of Example 1 For the rHA-SN-38 product prepared in Example 1, a pharmacodynamic experiment was conducted in a subcutaneously transplanted tumor model of human Hep 3B cells (ATCC HB-8064 cells) in BALB / c nude mice, and safety was evaluated at the same time.

[0334] Selected eligible tumor-bearing BALB / c nude mice (5 mice per group) were administered the rHA-SN-38 product (30 mg / kg) of Example 1 once a week for 6 consecutive weeks via tail vein injection. The animals' body weights were measured on days 13, 16, 20, 23, 27, 30, 34, 37, and 41 after administration. The results are shown in Figure 8.

[0335] The results show that the product of Example 1 (30 mg / kg) had no effect on the body weight of the animals, and had good safety and tumor-inhibiting efficacy.

[0336] Example 22: Investigation of the anti-breast tumor activity of rHA-SN-38 products For the rHA-SN-38 product prepared in Example 1, pharmacodynamic experiments were conducted in a subcutaneously transplanted tumor model of human triple-negative breast cancer MDA-MB-231 (ATCC: CRM-HTB-26™) in BALB / c nude mice to investigate the use of the composition of the present invention in cancer treatment.

[0337] The study was conducted according to two protocols. Protocol I: Twenty-one eligible tumor-bearing BALB / c nude mice were selected and randomly divided into three groups with seven mice per group. They were administered sterile water for injection, commercially available irinotecan hydrochloride injection (60 mg / kg), and rHA-SN-38 product (15 mg / kg), respectively. The mice were administered the drugs via tail vein injection three times a week for three consecutive weeks. The first day of administration was designated as day 0, and the tumor burden of each animal was measured on this day. During the administration period, the animals were observed daily for general clinical signs, and their body weights and tumor burdens were measured twice a week.

[0338] The experimental results are shown in Table 26. The rHA-SN-38 product of the present invention showed a highly significant tumor-inhibiting effect in a subcutaneously transplanted tumor model of human triple-negative breast cancer MDA-MB-231 in BALB / c nude mice, and was significantly superior to commercially available irinotecan hydrochloride injection. After three doses, the tumor inhibition rate of the rHA-SN-38 product (15 mg / kg) was 95%, while the tumor inhibition rate of the commercially available irinotecan hydrochloride injection (60 mg / kg) was 71%.

[0339] [Table 28]

[0340] Protocol II: Forty-two eligible tumor-bearing BALB / c nude mice were selected and randomly divided into six groups with seven mice per group. They were administered sterile water for injection, commercially available irinotecan hydrochloride injection (60 mg / kg, 80 mg / kg), rHA (220 mg / kg), and rHA-SN-38 products (1.67 mg / kg, 5 mg / kg, 15 mg / kg), respectively. The mice were administered the drugs via tail vein intravenous injection once weekly for three consecutive weeks. The first day of administration was designated as day 0, and tumor burden for each animal was measured on this day. During the treatment period, the animals were observed daily for general clinical signs, and body weight and tumor burden were measured twice weekly.

[0341] The experimental results are shown in Figure 9. As shown in Figure 9, the various doses of rHA-SN-38 preparation used showed a dose-dependent significant benefit in tumor inhibition.

[0342] As shown by the above experimental results, the rHA-SN-38 product of the present invention has a low administration dose, a high tumor inhibition rate, and a significantly better therapeutic effect than the commercially available irinotecan hydrochloride injection, and has an excellent therapeutic effect against human triple-negative breast cancer.

[0343] Example 23: Investigation of the anti-colon cancer activity (HT-29) of rHA-SN-38 product The rHA-SN-38 lyophilized powder product prepared in Example 2 was reconstituted and subjected to pharmacodynamic experiments in a subcutaneously transplanted tumor model of human colon carcinoma HT-29 (ATCC:HTB 3B™) in BALB / c nude mice to investigate the use of the composition of the present invention in cancer treatment.

[0344] The study was conducted according to two protocols. Protocol I: Twenty-one eligible tumor-bearing BALB / c nude mice were selected and randomly divided into three groups with seven mice per group. They were administered sterile water for injection, commercially available irinotecan hydrochloride injection (60 mg / kg), and rHA-SN-38 product (30 mg / kg), respectively. The mice were administered the drugs via tail vein injection once a week for three consecutive weeks. The first day of administration was designated as day 0, and the tumor burden of each animal was measured on this day. During the administration period, the animals were observed daily for general clinical signs, and their body weights and tumor burdens were measured twice a week.

[0345] The experimental results are shown in Table 27. The rHA-SN-38 product of the present invention (30 mg / kg) showed a highly significant tumor-inhibiting effect in a subcutaneously transplanted human colon cancer HT-29 tumor model in BALB / c nude mice, and was significantly superior to commercially available irinotecan hydrochloride injection (60 mg / kg). After four doses, the tumor inhibition rate of the rHA-SN-38 product (30 mg / kg) was 72%, while the tumor inhibition rate of the commercially available irinotecan hydrochloride injection (60 mg / kg) was 47%.

[0346] [Table 29]

[0347] Protocol II: Fifteen eligible tumor-bearing BALB / c nude mice were selected and randomly divided into three groups with five mice per group. They were administered sterile water for injection, commercially available irinotecan hydrochloride injection (60 mg / kg, 80 mg / kg), and rHA-SN-38 product (3.3 mg / kg, 10 mg / kg, 30 mg / kg), respectively. The mice were administered the drugs via tail vein intravenous injection once weekly for four consecutive weeks. The first day of administration was designated as day 0, and tumor burden for each animal was measured on this day. During the administration period, the animals were observed daily for general clinical signs, and body weight and tumor burden were measured twice weekly.

[0348] As shown in Figure 10, various doses of the rHA-SN-38 product demonstrated a significant dose-dependent benefit in tumor inhibition.

[0349] As shown by the above experimental results, the rHA-SN-38 product of the present invention had a low administration dose, a high tumor inhibition rate, and showed significantly better therapeutic effects than commercially available irinotecan hydrochloride injection, demonstrating excellent therapeutic effects against human colon cancer.

[0350] Example 24: In vivo pharmacodynamic testing on MDA-MB-231 tumors the purpose: The antitumor activity of various doses of the HSA-SN-38 product of Example 19 (administered after reconstitution in vehicle) in a subcutaneous xenograft model of the human triple-negative breast cancer cell line MDA-MB-231 (ATCC: CRM-HTB-26™) in BALB / c nude mice was evaluated and compared with the antitumor activity of the commercially available injectable irinotecan hydrochloride (CPT-11).

[0351] 1. Experimental Design

[0352] [Table 30]

[0353] 2. Experimental Method Forty-nine Balb / c female nude mice (6-8 weeks old) were selected and inoculated with MDA-MB-231 tumor masses into the right scapula of the nude mice. Sixteen days after tumor mass inoculation, the animals were randomly divided into groups (7 animals per group) and administered intravenous injection of the drug via the tail vein. One week after the final administration, tumors were removed from all mice and weighed.

[0354] 3. Experimental observation and data collection After tumor cell inoculation, in addition to observing tumor growth, the effects of treatment on the behavior of the animals were also detected, including the activity of the test animals, food and water intake, body weight changes (body weight was measured twice a week), and any abnormalities in the eyes, fur, and other conditions. Clinical symptoms observed during the experiment were recorded in raw data. Tumor volume was calculated using the following formula: Tumor volume (mm 3 )=1 / 2×(a×b 2 ) (a represents the major axis and b represents the minor axis). Dosing was stopped when animals lost more than 15% of their body weight (BWL > 15%) and resumed when body weight loss returned to less than 10%. When animals lost more than 20% of their body weight, they were euthanized in accordance with animal welfare regulations.

[0355] 4. Evaluation criteria for treatment effect The relative tumor growth rate (T / C (%)) represents the ratio of the relative tumor burden or tumor weight in percent between the treatment group and the control group at a single time point and was calculated by the following formula: T / C(%)=T RTV / C RTV ×100% In the formula, T RTV is the mean RTV of the treatment group; C RTV is the mean RTV of the vehicle control group; where: RTV=V t / V0, where V0 is the tumor burden of the animal at the time of grouping, and V t was the tumor burden of the animals after treatment; Or, T / C%=T TW / C TW ×100% In the formula, T TW is the mean tumor weight of the treatment group at the end of the experiment; C TW is the mean tumor weight of the vehicle control group at the end of the experiment. The relative tumor inhibition rate (TGI (%)) was calculated using the following formula: TGI(%)=(1-T / C)×100% where T and C were the relative tumor volume (RTV) or tumor weight (TW) of the treated and control groups, respectively, at a particular time point.

[0356] 5.Statistical analysis In the experiment, tumor mean values from various groups were compared using one-way analysis of variance. Homogeneous variance analysis indicated significant differences in F values, and multiple comparisons were performed using one-way analysis of variance followed by Dunnett's T3 (heteroscedasticity) method. All data were analyzed using SPSS 17.0. p<0.05 was considered to indicate a significant difference.

[0357] 6. Experimental Results 6.1 Weight change: As shown in Figure 11. 6.2 Tumor burden change: As shown in Figure 12. 6.3 Evaluation index of antitumor efficacy Table 29 below shows the endpoints of tumor-inhibitory efficacy of agents such as HSA-SN-38 and CPT-11 in the MDA-MB-231 xenograft model.

[0358] [Table 31]

[0359] 7. Conclusion High-dose HSA-SN-38 (15 mg / kg) showed a clear tumor-inhibiting effect, which was superior to that of CPT-11. At the end of the experiment (day 33 after tumor inoculation), after three doses, the high-dose HSA-SN-38 group had a tumor volume of 128 mm 3 The results showed an RTV of 0.89, a T / C value of 5%, and p=0.015. The intermediate dose of HSA-SN-38 (5 mg / kg) also showed a certain inhibitory effect on tumor growth, but this was not significantly different from the vehicle group (p=0.130). No significant antitumor activity was observed in the low-dose (1.67 mg / kg) HSA-SN-38-treated group. The tumor weight analysis results were substantially consistent with the tumor volume analysis results.

[0360] The effects on body weight changes of tumor-bearing mice in each group are shown in Figure 11. During the experiment, one animal died in the high-dose (80 mg / kg) CPT-11 treatment group, and the remaining six animals showed no significant weight loss after the second administration. At the end of the experiment, body weight increased by 1.95% compared to pre-administration weight. In the low-dose (60 mg / kg) CPT-11 treatment group, no animals died, and after the final administration, body weight increased by 2.41% compared to pre-administration weight. In the three HSA-SN-38 dose groups, no animals died, and no other abnormal toxic reactions were observed. After the final administration, body weight increased to various degrees compared to pre-administration weight, with the weight gains in the high-, medium-, and low-dose groups being 3.70%, 7.63%, and 4.85%, respectively. The vehicle and A01S control groups showed the most significant weight gain, with body weights increasing by 10.84% and 9.04%, respectively, compared to pre-treatment weights at the end of the experiment.

[0361] In summary, HSA-SN-38 at a dose of 15 mg / kg showed significant effects in inhibiting tumor growth in the MDA-MB-231 human breast cancer model. Similarly, CPT-11 at a dose of 80 mg / kg showed significant effects in inhibiting tumor growth, but the effects were weaker than those of the high-dose HSA-SN-38 group, with one animal dying after the second drug administration. Overall, HSA-SN-38 exhibited more potent antitumor activity than CPT-11 and was well tolerated; no animals died during the experiment, and no other toxic reactions were observed.

[0362] Example 25: In vivo pharmacodynamic testing on HCT116 tumors the purpose: The antitumor activity of HSA-SN-38 (administered after reconstitution in a vehicle) prepared in Example 20 was examined in a subcutaneous xenograft model of the human colon cancer cell line HCT116 (ATCC CCL-247) in BALB / c nude mice, and compared with that of commercially available injectable irinotecan hydrochloride (CPT-11).

[0363] 1. Experimental Design

[0364] [Table 32]

[0365] 2. Experimental Method A well-preserved HCT116 tumor mass of 20-30 mm 3 The tumor masses were cut into small tumor masses of 121 mm and inoculated into the right scapula of a total of 70 nude mice. 15 days after tumor mass inoculation, the average tumor volume was approximately 121 mm. 3 When the tumor burden reached 100 mg / kg, mice with excessively small or large tumor burdens were screened out. The remaining 49 mice were randomly divided into seven groups (7 mice per group) based on tumor burden and administered once a week (via tail vein injection) for a total of 4 weeks. The therapeutic effect was evaluated based on the relative tumor inhibition rate (TGI), and safety was evaluated based on the weight change and death of the animals. One week after the final administration, tumors were removed from all mice, weighed, and photographed.

[0366] 3.Statistical analysis In the experiment, tumor mean values from various groups were compared using one-way analysis of variance. Homogeneous variance analysis indicated significant differences in F values, and multiple comparisons were performed using one-way analysis of variance followed by Dunnett's T3 (heteroscedasticity) method. All data were analyzed using SPSS 17.0. p<0.05 was considered to indicate a significant difference.

[0367] 4. Experimental Results 4.1 Weight change: As shown in Figure 13. 4.2 Changes in tumor burden The changes in tumor burden in the various groups are shown in Table 31 and FIG.

[0368] [Table 33]

[0369] 4.3 Evaluation index of antitumor efficacy Table 32 shows the endpoints of anti-tumor efficacy of agents such as HSA-SN-38 and CPT-11 in the HCT116 xenograft model.

[0370] [Table 34]

[0371] 5. Conclusions and Discussion In the experiment, it was observed that high-dose HSA-SN-38 (30 mg / kg) showed significant tumor-inhibitory effects, which were superior to those of CPT-11. At the end of the experiment (43 days after tumor inoculation (D43)), after four doses, the high-dose HSA-SN-38 group had a tumor volume of 113 mm 3 The intermediate dose of HSA-SN-38 (10 mg / kg) showed antitumor activity equivalent to that of the low dose (60 mg / kg) CPT-11 group, with a tumor volume of 433 mm. 3 The RTV was 3.75, and the T / C value was 30%, which was significantly different from the vehicle control group (p=0.018). The results of tumor weight analysis were substantially consistent with the results of tumor volume analysis.

[0372] During the experiment, no animals died in any of the treatment groups. However, the body weights of animals in various groups, including the vehicle group, decreased to different degrees. At the end of the experiment, the weight loss of animals compared to pre-administration weights was 1.83% in the vehicle group, 5.41% in the A01S group, 16.00% and 14.41% in the high-dose and low-dose (80 mg / kg and 60 mg / kg) CPT-11 groups, respectively, and 11.84%, 13.85%, and 12.57% in the three dose (30 mg / kg, 10 mg / kg, and 3.3 mg / kg) HSA-SN-38 groups, respectively. Considering that the body weight of animals in the vehicle and A01S groups in the experiment decreased to various degrees, and that no obvious weight loss was observed in animals in the HSA-SN-38-treated groups in pharmacological experiments in other models, it was thought that the weight loss of animals in each HSA-SN-38-treated group in the experiment might be related to the cachexia characteristics of the HCT116 model.

[0373] In summary, HSA-SN-38 (at doses of 30 mg / kg and 10 mg / kg) showed significant efficacy in inhibiting tumor growth in a human colon cancer model, was superior to the positive control CPT-11, and was well tolerated; no animals died during administration and no other toxic reactions were observed in the study.

[0374] Example 26: In vivo pharmacodynamic testing in the SKOV-3 human ovarian cancer model the purpose: The antitumor activity of various doses of HSA-SN-38 (administered after reconstitution in vehicle) of Example 19 was examined in a subcutaneous xenograft model of the human ovarian cancer cell line SKOV-3 (ATCC HTB 77) in NU / NU nude mice, and compared with that of the commercially available injectable irinotecan hydrochloride (CPT-11).

[0375] 1. Experimental Design

[0376] [Table 35]

[0377] 2. Experimental Method SKOV-3 tumor masses were subcutaneously inoculated into 80 BALB / c nude mice. On day 14 after inoculation, 56 tumor-bearing mice were selected and evenly divided into seven groups with eight mice per group. Mice were administered the drug via tail vein injection once a week for a total of four weeks. The therapeutic efficacy was evaluated based on the relative tumor inhibition rate (TGI), and safety was assessed based on the weight change and death of the animals.

[0378] 3.Statistical analysis In the experiment, tumor mean values from various groups were compared using one-way analysis of variance. Homogeneous variance analysis indicated significant differences in F values, and multiple comparisons were performed using one-way analysis of variance followed by Dunnett's T3 (heteroscedasticity) method. All data were analyzed using SPSS 17.0. p<0.05 was considered to indicate a significant difference.

[0379] 4. Experimental Results 4.1 Weight change: As shown in Figure 15. 4.2 Changes in tumor burden The changes in tumor burden in the various groups are shown in Table 34 and FIG.

[0380] [Table 36]

[0381] 4.3 Evaluation index of antitumor efficacy Table 35 shows the evaluation indexes of the antitumor efficacy of HSA-SN-38 and CPT-11 in the SKOV-3 xenograft model.

[0382] [Table 37]

[0383] High-dose HSA-SN-38 (30 mg / kg) showed significant tumor-inhibitory effects, superior to low-dose CPT-11 (60 mg / kg). At the end of the experiment (day 42 after tumor inoculation (D42)), after four doses, the high-dose HSA-SN-38 group had a tumor volume of 445 mm. 3 The tumor weight analysis was substantially consistent with the tumor volume analysis.

[0384] During the experiment, six animals died in the high-dose (80 mg / kg) CPT-11 treatment group, and none died in the other groups. At the end of the experiment, the animals' body weights increased to various degrees. The body weights of animals in the high-, medium-, and low-dose HSA-SN-38 groups increased by 5.60%, 5.65%, and 7.03%, respectively. The body weights of animals in the low-dose CPT-11 group increased by 6.97% compared to pre-treatment weights, and the body weights of the two surviving animals in the high-dose group increased by 11.94%. The weight gain of animals in the vehicle and A01S control groups was most significant; at the end of the experiment, body weights increased by 8.86% and 11.99%, respectively, compared to pre-treatment weights.

[0385] Conclusion: HSA-SN-38 at a dose of 30 mg / kg showed significant efficacy in inhibiting tumor growth in the SKOV-3 human ovarian cancer model, was superior to CPT-11, and was well tolerated, with no animal deaths and no other toxic reactions observed during the experiment. Example 27: In vivo pharmacodynamic testing in the SW620 human colon cancer model

[0386] the purpose: The antitumor activity of various doses of rHA-SN-38 prepared in Example 4 was examined in a subcutaneous xenograft model of human colon cancer cell line SW620 (ATCC: CCL-227) in BALB / c nude mice, and compared with that of commercially available injectable irinotecan hydrochloride (CPT-11).

[0387] 1. Experimental Design

[0388] [Table 38]

[0389] 2. Experimental Method SW620 tumor masses were subcutaneously inoculated into 75 BALB / c nude mice. On day 13 after inoculation (D13), 49 tumor-bearing mice were selected and evenly divided into seven groups with seven mice per group. Mice were administered the drug via tail vein injection once a week for a total of 4 weeks. The therapeutic efficacy was evaluated based on the relative tumor inhibition rate (TGI), and safety was assessed based on the weight change and death of the animals.

[0390] 3.Statistical analysis In the experiment, tumor mean values from various groups were compared using one-way analysis of variance. Homogeneous variance analysis indicated significant differences in F values, and multiple comparisons were performed using one-way analysis of variance followed by Dunnett's T3 (heteroscedasticity) method. All data were analyzed using SPSS 17.0. p<0.05 was considered to indicate a significant difference.

[0391] 4. Experimental Results 4.1 Weight change: As shown in Figure 17. 4.2 Changes in tumor burden The changes in tumor burden in the various groups are shown in Table 37 and FIG.

[0392] [Table 39]

[0393] 4.3 Evaluation index of antitumor efficacy Table 38 shows the endpoints of antitumor efficacy of rHA-SN-38 and CPT-11 in the SW620 xenograft model.

[0394] [Table 40]

[0395] High- and intermediate-dose HSA-SN-38 (30 mg / kg and 10 mg / kg) showed significant tumor-inhibitory effects, superior to those of CPT-11. At the end of the experiment (day 41 after tumor inoculation (D41)), after four doses, the high- and intermediate-dose rHA-SN-38 groups each had tumor volumes of 21 mm 3and 101mm 3 The RTV values were 0.17 and 0.81, and T / C values were 1% and 5%, which were significantly different from the vehicle group (p values were 0.040 and 0.047, respectively). The high-dose and low-dose CPT-11 groups (80 mg / kg and 60 mg / kg) showed similar efficacy, no clear dose-effect relationship, and no significant difference from the vehicle group (p values were 0.202 and 0.133, respectively). At D41, the tumor burden in the low-dose (3.3 mg / kg) rHA-SN-38 group was not significantly different from that in the vehicle group (p=0.231). The tumor weight analysis results were substantially consistent with the tumor volume analysis results.

[0396] No animals died in any group, and no other abnormal toxic reactions were observed. At the end of the experiment, the body weights of animals in the high-dose and medium-dose rHA-SN-38 groups had increased by 2.03% and 4.01%, respectively, compared to pre-treatment body weights. At the end of the experiment, the body weights of animals in the other groups had decreased to various degrees compared to pre-treatment body weights. The decreases were 5.81% and 0.99% in the high-dose and low-dose CPT-11 groups, respectively, 0.33% in the low-dose rHA-SN-38 group, and 5.83% and 4.39% in the vehicle and A01S control groups, respectively.

[0397] In summary, rHA-SN-38 at doses of 30 mg / kg and 10 mg / kg significantly inhibited tumor growth in the SW620 human colon cancer model, and was superior to CPT-11 at doses of 80 mg / kg and 60 mg / kg. Furthermore, the body weight of animals in the two rHA-SN-38-treated groups increased significantly, while the body weight of animals in the two CPT-11-treated groups decreased or increased only slightly. Overall, rHA-SN-38 exhibited significantly more potent antitumor activity than CPT-11 and was well tolerated; no animals died during the experiment, and no other toxic reactions were observed.

[0398] Example 28: In vivo pharmacodynamic testing on Hep3B human liver cancer model the purpose: The antitumor activity of rHA-SN-38 prepared in Example 5 was examined in a subcutaneous xenograft model of human hepatocellular carcinoma cell line Hep3B (ATCC-8064) in BALB / c nude mice, and compared with that of commercially available injectable irinotecan hydrochloride (CPT-11).

[0399] 1. Experimental Design

[0400] [Table 41]

[0401] 2. Experimental Method Hep3B tumor masses were subcutaneously inoculated into 75 BALB / c nude mice. On day 13 after inoculation (D13), 49 tumor-bearing mice were selected and evenly divided into seven groups with seven mice per group. Mice were administered the drug via tail vein injection once a week for a total of 4 weeks. The therapeutic efficacy was evaluated based on the relative tumor inhibition rate (TGI), and safety was assessed based on the weight change and death of the animals.

[0402] 3.Statistical analysis In the experiment, tumor mean values from various groups were compared using one-way analysis of variance. Homogeneous variance analysis indicated significant differences in F values, and multiple comparisons were performed using one-way analysis of variance followed by Dunnett's T3 (heteroscedasticity) method. All data were analyzed using SPSS 17.0. p<0.05 was considered to indicate a significant difference.

[0403] 4. Experimental Results 4.1 Weight change: As shown in Figure 19. 4.2 Tumor burden change: As shown in Figure 20.

[0404] conclusion The test article rHA-SN-38 at doses of 30 mg / kg and 10 mg / kg significantly inhibited tumor growth in a Hep3B human liver cancer model, with the same antitumor levels as CPT-11 at a dose of 80 mg / kg. CPT-11 at a dose of 60 mg / kg exhibited antitumor activity similar to the tumor-inhibitory effect of HSA-SN-38 at a dose of 3.33 mg / mL. rHA-SN-38 was well tolerated, with no animal deaths and no other toxic reactions observed during the study.

[0405] Example 29: Effect of Addition of Various Surfactants to SN-38 Formulations Preparation process: 1) Prepare a mixed solvent of EtOH / CHCl3 with a volume ratio of 3 / 7; 2) Take 300 mg of SN-38, 300 mg of cholesterol, and the amount of surfactant shown in Table 40 below, add 30 mL of the mixed solvent from step 1), and dissolve completely under heating to obtain a drug solution; 3) A total volume of 370 mL of an aqueous HSA solution was prepared using deionized water as the aqueous phase, so that the total content of HSA in the aqueous phase was 3 g; 4) Shear dispersion: Mix the drug solution from step 2) with the aqueous phase from step 3) and perform shear dispersion for 10 minutes to obtain a crude emulsion; 5) The crude emulsion is transferred to a high-pressure homogenizer and homogenized under a pressure of 1300-1500 bar for 3-8 times; 6) Transfer the homogenized product into a flask; 7) Rotary evaporation was carried out at 40°C to 45°C for 4 to 6 minutes to remove chloroform from the system; 8) EtOH in the system was removed through liquid exchange with a tangential flow ultrafiltration (TFF) system (Millipore); 9) Filtration was performed through a 0.2 μm PES syringe filter membrane. 10) The resulting product was stored in a refrigerator at 4°C. During preparation, the particle size of the samples was detected separately after shear dispersion, after high-pressure homogenization, and before and after filtration.

[0406] [Table 42]

[0407] The results show that adding various amounts of surfactant to the organic phase affected the number of homogenizations and particle size during the preparation process, and that Span 20 was superior to other surfactants or no surfactant in reducing the number of homogenizations and reducing the particle size of the nanoparticles.

[0408] Example 30: Effect of Addition of Span 20 on Filtration Flux of SN-38 Formulations To further investigate the effect of Span 20 on the preparation process, the particle size and filtration flux in the preparation step of SN-38 formulation were compared between the cases where Span 20 was added and the case where Span 20 was not added. The preparation process and the amount of Span 20 added were the same as in Example 29. The results are shown in Table 41.

[0409] [Table 43]

[0410] The results show that the product prepared using the process that added Span 20 had smaller particle size and a higher filtration flux through a PES syringe filter compared to the formulation prepared without Span 20.

[0411] Furthermore, disintegration experiments on the two samples were carried out according to the method described in item 9 of the preamble of the Examples. Specifically, the samples were diluted with 1x PBS at pH 7.4. The particle size of the samples at various dilution ratios was measured to investigate at which dilution ratio the nanoparticles disintegrated and the SN-38 raw material precipitated. The greater the dilution ratio, the better the stability of the nanoparticles.

[0412] The results of the disintegration experiment are shown in Figures 21 and 22. The results show that, with or without Span 20, the two samples remained stable when diluted to 0.1 μg / mL; when subsequently diluted to 10 μg / mL, the particle sizes of the two samples increased significantly, indicating that the nanoparticles had disintegrated. After adding Span 20, the particle size change after disintegration became smaller and more stable.

[0413] Furthermore, the morphologies of the two samples were observed using a cryogenic transmission electron microscope according to the method described in item 11 of the preamble of the Examples. The electron microscope observation results are shown in Figures 23 and 24. The results showed that the API in the two samples was in two different states: nanocrystals and vesicles, with more vesicles and fewer nanocrystals in the sample containing Span 20. SN-38 in the form of vesicles became a fast-acting API after entering the human body, indicating that the addition of Span 20 was useful in better maintaining the nanoparticle size while enabling the formulation to exert its efficacy in the body as quickly as possible.

[0414] Example 31: Effect of adding varying amounts of Span 20 to SN-38 formulations Preparation process: 1) An organic solvent system was prepared as shown in Table 42 below; 2) SN-38, cholesterol, and Span 20 were weighed as shown in Table 42, and 30 mL of the organic solvent from step 1) was added and completely dissolved to obtain a drug solution; 3) A total volume of 370 mL of an HSA aqueous solution was prepared using deionized water as the aqueous phase, so that the total content of HSA in the aqueous phase was 3 g; 4) Shear dispersion: Mix the drug solution from step 2) with the aqueous phase from step 3) and perform shear dispersion for 10 minutes to obtain a crude emulsion; 5) The crude emulsion is transferred to a high-pressure homogenizer and homogenized 3 to 5 times under a pressure of 1300 to 1500 bar; 6) Transfer the homogenized product into a flask; 7) Rotary evaporation was carried out at 40°C to 45°C for 4 to 6 minutes to remove chloroform from the system; 8) DMSO or EtOH in the system is removed through liquid exchange by TFF; 9) Filtration was performed through a 0.2 μm PES syringe filter membrane, and parameters such as particle size, API filtration recovery rate, and drug loading of the sample were detected before and after filtration; 10) The samples were stored in a refrigerator at 4°C. The results are shown in Table 42.

[0415] [Table 44]

[0416] The results show that for various SN-38 and cholesterol content ratios, adding Span 20 in the preparation process can reduce the particle size of nanoparticles, improve filtration efficiency and filtration flux, and increase filtration recovery. Single-factor comparison shows that Span 20 can increase the drug loading and API recovery.

[0417] Example 32: Effect of Addition of Span 20 on the Stability of SN-38 Formulations The effect of Span 20 on the formation of albumin multimers in solution was observed by measuring the content of human serum albumin multimers in SN-38 formulations.

[0418] Experimental Method: 1) Formulations 11 and 12 from Example 31 were filled into vials and freeze-dried under reduced pressure to obtain freeze-dried formulations of HSA-SN-38 nanoparticles. After 14 hours of storage, the formulations were diluted with deionized water to the same SN-38 concentration as before freeze-drying, and stored under high temperature, high humidity, or strong light conditions for 5 or 10 days for use as samples to be measured. 2) The content of human serum albumin multimers in SN-38 formulations prepared by various methods was measured using SEC-HPLC. 5 μl of the prepared sample was taken for detection, and the chromatographic conditions were as shown in Table 43.

[0419] [Table 45]

[0420] The results are shown in Table 44 below.

[0421] [Table 46]

[0422] The results show that Formulation 11 with added Span 20 showed no obvious change in albumin multimer content compared to Formulation 12 without added Span 20, indicating that Span 20 can inhibit albumin aggregation. Therefore, Span 20 can substantially prevent the aggregation of albumin in solution, thereby extending the shelf life of the drug, and does not exhibit the immunogenicity caused by albumin multimers.

[0423] Example 33: Effect of albumin content on SN-38 formulations SN-38 formulations were prepared according to the formula shown in Table 45 by the method of Example 29, except that Span 20 was not added to Formulation 13, and 6 g and 12 g of HSA were included in Formulations 15 and 16, respectively. Particle size was measured during the steps of preparing the SN-38 formulations, and API filtration recovery was also measured to investigate the effect of HSA content on the formulation. The results are shown in Table 46.

[0424] [Table 47]

[0425] [Table 48]

[0426] The results show that increasing the amount of albumin in the formulation increases the particle size of the sample, significantly reduces the filtration flux, and reduces the API filtration recovery. It is appropriate to control the ratio of HSA to API at 20:1 or less. If the ratio exceeds this ratio, especially if it is 40:1 or more, the nanoparticles will not only have excessively large particle sizes, become unstable, and be prone to aggregation and immunogenicity, but will also be difficult to filter, resulting in low API recovery and making large-scale production impossible.

[0427] Example 34: In vivo pharmacodynamic testing in the SW620 human colon cancer model the purpose: The antitumor activity and safety of SN-38 formulations containing various amounts of Span 20 and HSA prepared in Example 31 were investigated in a subcutaneous xenograft model of the human colon cancer cell line SW620 (ATCC: CCL-227) in BALB / c nude mice, and were compared with those of commercially available injectable irinotecan hydrochloride (CPT-11).

[0428] [Table 49]

[0429] cell culture In vitro monolayer cultures of SW620 cells were grown in 1640 medium supplemented with 10% heat-inactivated fetal bovine serum and agar at 37°C in an incubator under 5% CO2 in air, and were subcultured twice weekly with 0.25% pancreatin. During the exponential growth phase, cells were harvested, counted, and inoculated.

[0430] Tumor cell inoculation and tumor block passaging SW620 tumor cells 5.0 × 10 6 The cells were suspended in 0.1 mL of PBS and inoculated into the right scapula of five nude mice (P1 generation). 3After tumor growth, the tumor-bearing mice were anesthetized with CO2 and euthanized. The tumor masses were obtained and the surrounding necrotic tissue was removed. The tumor masses were then cut into 20-30 mm pieces. 3 The tumor mass was cut into small pieces and inoculated into the right scapula of a new batch of nude mice (P2 generation). A total of 40 mice were inoculated.

[0431] Tumor mass inoculation, grouping, and administration In the experiment, the antitumor activity of the test product was evaluated using P2 generation tumor tissue. Seven days after tumor mass inoculation, the average tumor volume was approximately 159 mm 3 When the tumor burden reached 100 mg / kg, mice with excessively small or large tumor burdens were screened out, and the remaining 36 mice were randomly divided into groups according to tumor burden, and administration was initiated. The results are shown in Figures 25 to 27 and Table 48.

[0432] [Table 50]

[0433] The results show that formulations with various amounts of Span 20 and formulations without Span 20 exhibited better antitumor activity than CPT-11.

[0434] Example 35: Effect on formulation of using EtOH / CH2Cl2 / CHCl3 mixed solvent instead of EtOH / CHCl3 mixed solvent 1. Preparation process: 1) Prepare a mixed solvent of EtOH / CH2Cl2 / CHCl3 according to the volume ratio shown in Table 49 below; 2) Take 300 mg of SN-38, 300 mg of cholesterol, and 18 mg of Span 20, add 30 mL of the mixed solvent from step 1), and dissolve completely under heating to obtain a drug solution; 3) A total volume of 370 mL of an HSA aqueous solution was prepared using deionized water as the aqueous phase, so that the total content of HSA in the aqueous phase was 3 g; Shear dispersion: Mix the drug solution from step 2) with the aqueous phase from step 3) and perform shear dispersion for 10-15 minutes to obtain a crude emulsion; 4) The crude emulsion was transferred to a high-pressure homogenizer and homogenized five times under a pressure of 1300-1500 bar, and the homogenized sample was transferred to a flask; 5) Rotary evaporation at 40°C to 45°C for 4 to 6 minutes; 6) EtOH in the system was removed through liquid exchange by TFF; 7) Filtration was performed through a 0.2 μm PES syringe filter membrane, and parameters such as particle size, API filtration recovery rate, and drug loading of the sample were detected before and after filtration; 8) The samples were stored in a refrigerator at 4°C.

[0435] [Table 51]

[0436] 2.Results: After adjusting the organic solvent system to EtOH / CH2Cl2 / CHCl3 = 6 / 7 / 7, the amount of residual CHCl3 was significantly reduced, and at the same time, the approximate quantification level of residual CH2Cl2 was also low (2 μg per mg of API). Based on the amount of residual CHCl3 in the filtered liquid and the maximum daily exposure of CHCl3 (600 μg), the maximum clinical dose was calculated to be 100 mg / m 2 The dose limitation caused by residual CHCl3 was significantly reduced. However, the crude product prepared using the EtOH / CHCl / CHCl3 mixed solvent had lower solubility of SN-38 in CHCl2 and rapid crystal precipitation when the organic phase was dispersed in the aqueous phase, resulting in larger particle size, lower API filtration recovery, and lower drug loading.

[0437] After adjusting the amount of CHCl to reach EtOH / CHCl / CHCl=6 / 4 / 10, the particle size of the crude product decreased, and parameters such as API filtration recovery were consistent with those of the product prepared using EtOH / CHCl mixed solvent, but the level of residual CHCl was still significantly reduced.

[0438] Example 36: Large-scale preparation of HSA-SN-38 product spiked with SP20 Preparation process: 1) Prepare 300 mL of mixed organic solvent as shown in Table 50 below; 2) Take 3 g of SN-38, 3 g of cholesterol, and 0.18 g of Span 20, add 30 mL of the mixed organic solvent from step 1), and dissolve completely under heating to obtain a drug solution; 3) Take 150 mL of 20% HSA solution and dilute it with 3550 mL of deionized water to obtain the aqueous phase; 4) Shear dispersion and homogenization: An in-line shearer (Fluke FDHS3 / 60) was connected in series with a high-pressure homogenizer (ATS, AH12-150) in continuous production mode, and the rotation speed of the in-line shearer was set at 8,000-10,000 rpm. The aqueous phase from step 3 and the organic phase from step 2 were pumped into the in-line shearer at a certain ratio, and then the drug solution was fed into the high-pressure homogenizer and homogenized five times under a pressure of 1,300-1,500 bar. 5) Evaporation: The chloroform and dichloromethane in the system were removed by evaporation in a falling film evaporator (evaporation tube temperature: 40°C to 45°C); 6) Removing residual EtOH in the system through liquid exchange with TFF; 7) Sucrose was added at a concentration of 80 mg / ml; 8) Filtration was carried out through a 0.2 μm bag filter, and parameters such as particle size, API filtration recovery rate, and drug loading of the sample were detected before and after filtration; 9) The samples were filled into vials at 15 mL per vial and then freeze-dried. Residual solvents in the freeze-dried samples were detected.

[0439] [Table 52]

[0440] The results show that the residual chloroform level of the product prepared using EtOH / CH2Cl2 / CHCl3 = 6 / 4 / 10 as the mixed organic solvent was significantly reduced, and the properties of the product prepared in the scaled-up production were consistent with those of the product prepared by small-scale batch.

[0441] Example 37: Effect of Addition of Span 20 on the Stability of HSA-SN-38 Formulations at Various Time Points The effect of Span 20 on the stability of albumin in solution (multimers, particle size, etc.) was investigated by measuring the content of human serum albumin multimers in SN-38 formulations.

[0442] Experimental Method: 1) Formulation 12 from Example 31 and Formulation 18 from Example 36 were filled into vials and freeze-dried under reduced pressure to obtain freeze-dried HSA-SN-38 formulations. Prior to the start of the stability experiment, the relevant samples were stored in a refrigerator (2°C to 8°C) for 0 days, 3 months, and 6 months, respectively. The samples were then diluted with deionized water to the same SN-38 concentration as before freeze-drying, and the moisture content, pH value, osmolality, particle size and particle size distribution, and content of multimers and cholesterol were then determined. The results are shown in Table 50 (Formulation 18) and Table 51 (Formulation 12). 2) The content of human serum albumin multimers in SN-38 formulations prepared by various methods was measured using SEC-HPLC. 5 μl of the prepared sample was directly taken for detection, and the chromatographic conditions were as shown in Table 43.

[0443] [Table 53]

[0444] [Table 54]

[0445] The above results indicate that the changes in particle size and albumin multimers in the HSA-SN-38 formulation to which Span 20 was added were reduced over time, and that the stability was improved.

[0446] Example 38: In vivo pharmacodynamic testing in the SKOV-3 human ovarian cancer model 1. Purpose of the experiment: The antitumor activity and safety of formulation 12 prepared in Example 31 and formulation 18 prepared in Example 36 were examined in a subcutaneous xenograft model of the human ovarian cancer cell line SKOV-3 (ATCC HTB 77) in BALB / c nude mice.

[0447] 2. Experimental Method SKOV-3 tumor masses were subcutaneously inoculated into 40 BALB / c nude mice. On day 13 after inoculation (D13), 18 tumor-bearing mice were selected and evenly divided into three groups with six mice per group. The mice were administered the drug via tail vein injection once a week for a total of four weeks, with the specific dosing regimen shown in Table 53. The therapeutic efficacy was evaluated based on the relative tumor inhibition rate (TGI), and safety was evaluated based on the weight change and death of the animals.

[0448] [Table 55]

[0449] 3.Statistical analysis In the experiment, tumor mean values from various groups were compared using one-way analysis of variance. Homogeneous variance analysis indicated significant differences in F values, and multiple comparisons were performed using one-way analysis of variance followed by Dunnett's T3 (heteroscedasticity) method. All data were analyzed using SPSS 17.0. p<0.05 was considered to indicate a significant difference.

[0450] 4. Experimental Results 4.1 Weight change: As shown in Figure 28. 4.2 Changes in tumor burden: As shown in Figures 29 and 30. 4.3 Table 54 shows the antitumor efficacy endpoints of HSA-SN-38 Formulations 12 and 18 in the SKOV-3 xenograft model.

[0451] [Table 56]

[0452] Conclusion: Formulations 12 and 18 at a dose of 30 mg / kg significantly inhibited tumor growth in the SKOV-3 human ovarian cancer model, ultimately resulting in substantial tumor disappearance. Furthermore, no animals died or other toxic reactions were observed during the experiment. The results indicate that the formulation containing Span 20 (Formulation 18) and the formulation without Span 20 (Formulation 12) exhibited comparable antitumor activity.

[0453] Example 39: In vivo pharmacodynamic testing in the HCT116 human colon cancer model 1. Purpose of the experiment: The antitumor activity and safety of Formulation 12 prepared in Example 31 and Formulation 18 prepared in Example 36 were examined in a subcutaneous xenograft model of the human colon cancer cell line HCT116 (ATCC CCL-247) in BALB / c nude mice.

[0454] 2. Experimental Method A well-preserved HCT116 tumor mass of 20-30 mm 3 The mice were cut into small tumor masses of 121 mm and inoculated into the right scapula of 40 mice. 15 days after tumor mass inoculation, the average tumor volume was approximately 121 mm. 3 When the tumor burden reached 100%, mice with excessively small or large tumor burdens were screened out. The remaining 18 mice were randomly divided into three groups (6 mice per group) according to tumor burden, and the drug was administered (via tail vein injection) once a week for a total of 4 weeks. The therapeutic effect was evaluated based on the relative tumor inhibition rate (TGI), and safety was evaluated based on the weight change and death of the animals. One week after the final administration, tumors were removed from all mice, weighed, and photographed.

[0455] [Table 57]

[0456] 3.Statistical analysis In the experiment, tumor mean values from various groups were compared using one-way analysis of variance. Homogeneous variance analysis indicated significant differences in F values, and multiple comparisons were performed using one-way analysis of variance followed by Dunnett's T3 (heteroscedasticity) method. All data were analyzed using SPSS 17.0. p<0.05 was considered to indicate a significant difference.

[0457] 4. Experimental Results 4.1 Weight change: As shown in Figure 31. 4.2 Tumor burden change: As shown in Figure 32 and Figure 33.

[0458] Conclusion: Formulations 12 and 18 at a dose of 10 mg / kg significantly inhibited tumor growth in the HCT116 human colon cancer model, resulting in tumor virtual disappearance. Furthermore, no animals died or other toxic reactions were observed during the experiment.

[0459] Equivalents and Incorporation by Reference The compositions, methods, and uses of the present application have been described herein with reference to certain preferred embodiments, but the present application should not be considered limited to the above-described embodiments, as certain variations will be apparent to those skilled in the art based on the disclosure set forth herein.

[0460] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. In this specification and claims, the singular forms "a," "an," and "the" include the plural forms as well, unless the context clearly dictates otherwise.

[0461] Furthermore, the methods of the present application are to some extent independent of the particular order of steps set forth herein, and the particular order of steps set forth in any claim should not be construed as a limitation on that claim.

[0462] All patents, patent applications, references, and publications cited herein are hereby incorporated by reference in their entirety.

Claims

1. A composition comprising SN-38, a lipid, albumin, and Span 20, said composition comprising nanoparticles, wherein the albumin encapsulates at least a portion of the SN-38 and optionally at least a portion of the lipid in said nanoparticles; Lipid:SN-38 is about (0.1-10):1 (w:w), about (0.5-6):1 (w:w), about (0.5-5):1 (w:w), about (0.5-3):1 (w:w), about (1-4):1 (w:w), about (1.2-4):1 (w:w), about (1.4-2):1 (w:w), about (1.5-2.5):1 (w:w), or about 1:1; albumin:SN-38 is about (1-100):1 (w:w), about (1-50):1 (w:w), about (3-25):1 (w:w), about (5-25):1 (w:w), about (5-20):1 (w:w), about (5-18):1 (w:w), about (6-15):1 (w:w), about (7-15):1 (w:w), about (6-12):1 (w:w), about (7-12):1 (w:w), about (9-11):1 (w:w), or about 10:1 (w:w); Span 20:SN-38 is about (3-60):100(w:w), about (4-60):100(w:w), about (5-60):100(w:w), about (6-60):100(w:w), about (7-55):100(w:w), about (8-50):100(w:w), about (10-45):100(w:w), about (12-40):100(w:w), about (14-35):100(w:w), about (15-30):100(w:w), about (16-25):100(w:w), or about (18-20):100(w:w); The composition, characterized in that the lipid is selected from cholesterol, cholesterol derivatives, cholesterol analogs, and fatty acid esters, and any combination of two or more thereof, the cholesterol derivative is selected from esters formed with cholesterol and an organic acid, and the cholesterol analog is selected from vitamin D2, vitamin D3, and combinations thereof.

2. 2. The composition of claim 1, wherein the albumin:lipid ratio is about (1-100):1 (w:w), such as about (2-20):1 (w:w), about (3-15):1 (w:w), about (5-10):1 (w:w), about 7:1 (w:w), or about 10:1 (w:w).

3. relative to the total amount of the SN-38, the lipid, and the albumin in the composition, the SN-38 content is about 1% to about 25% w / w; and / or the lipid content is from about 1% to about 35% w / w; and / or the albumin content is about 50 w / w% to about 98 w / w%; Or, the SN-38 content is about 3% to about 20% w / w; and / or the lipid content is about 2% to about 30% w / w; and / or the albumin content is about 55% to about 95% w / w; Or, the SN-38 content is about 3% to about 15%, about 4%, about 5%, about 6%, about 6.5%, about 7%, about 7.5%, about 8%, about 8.5%, about 9%, about 10%, about 11%, about 12%, about 13%, or about 14% w / w; and / or the lipid content is about 3% to about 30%, about 4%, about 5%, about 6%, about 7%, about 8%, about 8.5%, about 9%, about 9.5%, about 10%, about 10.5%, about 11%, about 11.5%, about 12%, about 12.5%, about 13%, about 13.5%, about 14%, about 15%, about 16%, about 17%, about 18%, about 19%, about 20%, about 21%, about 24%, about 26%, or about 28% w / w; and / or the albumin content is about 60% to about 94%, about 64% to about 93%, about 66% to about 92%, about 68% to about 91%, about 70% to about 90%, about 75% to about 90%, about 76%, about 77%, about 78%, about 79%, about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, or about 89%; and / or 2. The composition of claim 1, wherein the amount of Span 20 is about 0.03 w / w% to about 12 w / w%, about 0.06 w / w% to about 10 w / w%, 0.08 w / w% to about 9 w / w%, about 0.1 w / w% to about 8 w / w%, about 0.2 w / w% to about 7 w / w%, about 0.4 w / w% to about 6 w / w%, about 0.6 w / w% to about 5 w / w%, about 0.8 w / w% to about 4 w / w%, or about 1 w / w% to about 2 w / w% relative to the total amount of the SN-38, the lipid, the albumin, and the Span 20 in the composition.

4. Lipid:SN-38 is about (1-10):1(w:w), about (1-8):1(w:w), about (1-6):1(w:w), about (1-5):1(w:w), about (1-4.5):1(w:w), about (1-4):1(w:w), about (1.2-3.8):1(w:w), about (1.4-3.6):1(w:w), about (1.6-3.4):1(w:w), about (1.8-3.2):1(w:w), about (2-3):1(w:w), about (2.2-2.8):1(w:w), about (2.4-2.6):1(w:w), about 1:1(w:w), about 2.5:1(w:w), or about (0.8-1.8):1(w:w), about (0.9-1.7):1(w:w), or about (1-1.4):1; and / or albumin:SN-38 is about (5-40):1(w:w), about (5-<40):1(w:w), about (5-35):1(w:w), about (5-30):1(w:w), about (8-25):1(w:w), about (10-22.5):1(w:w), about (12.5-20):1(w:w), about (15-17.5):1(w:w), about (16-18):1(w:w), about 10:1(w:w), or about (9-21):1(w:w), about (9-20):1(w:w), about (11-18):1(w:w), or about (11.1-17.3):1(w:w); and / or Span 20:SN-38 is about (5-60):100(w:w), about (6-60):100(w:w), about (7-55):100(w:w), about (8-50):100(w:w), about (10-45):100(w:w), about (12-40):100(w:w), about (14-35):100(w:w), about (15-30):100 (w:w), about (16-25):100(w:w), about (18-20):100(w:w), or about (5-10):100(w:w), about (5-9):100(w:w), about (6-8.6):100(w:w), about (6-8):100(w:w), or about (6.5-7):100(w:w); and / or 2. The composition of claim 1, wherein the albumin:lipid ratio is about (1-40):1 (w:w), about (1-<40):1 (w:w), about (2-35):1 (w:w), about (3-15):1 (w:w), about (5-10):1 (w:w), about (6-8):1 (w:w), or about 7:1 (w:w), or about (6-21):1 (w:w), about (6.7-13):1 (w:w), about (7-13):1 (w:w), or about (11-12.7):1 (w:w).

5. relative to the total amount of the SN-38, the lipid, and the albumin in the composition, the SN-38 content is about 2% to about 16% w / w; and / or the lipid content is about 2% to about 35% w / w; and / or the albumin content is about 75% to about 96% w / w; Or, the SN-38 content is about 2.5% to about 15% w / w, about 4% to about 10% w / w, about 4.5% to about 9.5% w / w, about 5% to about 9% w / w, or about 7.5% to about 8% w / w; and / or The lipid content in the composition is about 2.5% to about 30% w / w, about 4% to about 12.5% w / w, about 4.5% to about 12% w / w, about 7% to about 10% w / w, or about 7.5% to about 8% w / w; and / or the albumin content in the composition is about 76 w / w% to about 95 w / w%, about 78 w / w% to about 93 w / w%, about 79 w / w% to about 91.5 w / w%, about 80 w / w% to about 90 w / w%, about 82 w / w% to about 89 w / w%, about 84 w / w% to about 88 w / w%, or about 84.5 w / w% to about 87.5 w / w%; Or, The content of SN-38 is about 3 w / w% to about 14 w / w%, about 3.5 w / w% to about 12 w / w%, about 4 w / w%, about 4.2 w / w%, about 4.5 w / w%, about 4.6 w / w%, about 4.8 w / w%, about 5 w / w%, about 5.5 w / w%, about 6 w / w%, about 6.5 w / w%, about 7 w / w%, about 7.5 %, about 7.6 w / w%, about 7.8 w / w%, about 8 w / w%, about 8.5 w / w%, about 9 w / w%, about 9.2 w / w%, about 9.5 w / w%, about 9.6 w / w%, about 9.8 w / w%, about 10 w / w%, about 10.5 w / w%, about 11 w / w%, or about 11.5 w / w%; and / or The lipid content is about 4 w / w% to about 25 w / w%, about 4.5 w / w% to about 20 w / w%, about 4.3 w / w%, about 4.5 w / w%, about 4.7 w / w%, about 5 w / w%, about 5.5 w / w%, about 6 w / w%, about 6.5 w / w%, about 6.7 w / w%, about 6.9 w / w%, about 7 w / w%, about 7.5 w / w%, about 7.6 w / w%, about 7.8 w / w%, about 8 w / w%, about 8.5 w / w%, about 9 w / w%, about 9.5 w / w%, about 10 w / w%, about 10.5 w / w %, about 11 w / w%, about 11.5 w / w%, about 12 w / w%, about 12.1 w / w%, about 12.3 w / w%, about 12.5 w / w%, about 13 w / w%, about 13.5 w / w%, about 14 w / w%, about 14.5 w / w%, about 15 w / w%, about 15.5 w / w%, about 16 w / w%, about 16.5 w / w%, about 17 w / w%, about 17.5 w / w%, about 18 w / w%, about 18.5 w / w%, about 19 w / w%, or about 19.5 w / w%; and / or The albumin content is about 78 w / w% to about 92 w / w%, about 79 w / w%, about 79.2 w / w%, about 79.4 w / w%, about 79.6 w / w%, about 79.8 w / w%, about 80 w / w%, about 81 w / w%, about 82 w / w%, about 83 w / w%, about 84 w / w%, about 84.3 w / w%, about 84.5 w / w%, about 84.7 w / w %, about 84.9 w / w%, about 85 w / w%, about 86 w / w%, about 87 w / w%, about 87.3 w / w%, about 87.5 w / w%, about 87.7 w / w%, about 87.9 w / w%, about 88 w / w%, about 89 w / w%, about 90 w / w%, about 91 w / w%, about 91.3 w / w%, or about 91.5 w / w%; and / or The content of the Span 20 relative to the total amount of the SN-38, the lipid, the albumin, and the Span 20 in the composition is about 0.14 w / w% to about 5 w / w%, about 0.2 w / w% to about 2.5 w / w%, about 0.22 w / w% to about 2.0 w / w%, about 0.24 w / w% to about 2 w / w%, about 0.26 w / w% to about 1.5 w / w%, about 0.28 w / w% to about 1.0 w / w%, or about 0.3 w / w%. ~about 0.9w / w%, about 0.32w / w%~about 0.8w / w%, about 0.34w / w%~about 0.7w / w%, about 0.36w / w%~about 0.6 w / w%, about 0.38 w / w% to about 0.58 w / w%, about 0.4 w / w% to about 0.56 w / w%, about 0.42 w / w% to about 0.54 w / w% , about 0.44 w / w% to about 0.52 w / w%, about 0.46 w / w%, about 0.48 w / w%, about 0.5 w / w%, about 0.2 w / w% to about 0.8 w / w%, about 0.24 w / w% to about 0.7 w / w%, about 0.26 w / w% to about 0.7 w / w%, about 0.3 w / w% to about 0.65 w / w%, about 0.36 w / w% to about 0.6 w / w%, about 0.4 w / w% to about 0.58 w / w%, about 0.44 w / w% to about 0.56 w / w%, about 0.48 w / w% to about 0.54 w / w%, or about 0.5 w / w% to about 0.52 w / w%.

6. the cholesterol derivative is selected from cholesteryl palmitate, cholesteryl caprylate, and combinations thereof; and / or 2. The composition of claim 1, wherein the fatty acid ester is selected from fatty acid glycerides, such as long chain fatty acid glycerides, including glyceryl stearate, e.g., glyceryl monostearate.

7. 2. The composition of claim 1, wherein the lipid is cholesterol.

8. cholesterol:SN-38 is about (1-6):1 (w:w), about (1.2-5):1 (w:w), about (1.4-4):1 (w:w), about 3:1 (w:w), about 2:1 (w:w), about 1:1 (w:w), or about (0.8-1.8):1 (w:w), about (0.9-1.7):1 (w:w), or about (1-1.4):1; and / or albumin:SN-38 is about (3-25):1 (w:w), about (4-20):1 (w:w), about (5-15):1 (w:w), about (6-12):1 (w:w), about (7-12):1 (w:w), about (9-11):1 (w:w), or about 10:1 (w:w), or about (9-21):1 (w:w), about (9-20):1 (w:w), about (11-18):1 (w:w), or about (11.1-17.3):1 (w:w); and / or albumin:cholesterol is about (2-20):1 (w:w), about (3-15):1 (w:w), about (5-10):1 (w:w), or about 7:1 (w:w), or about (6-21):1 (w:w), about (6.7-13):1 (w:w), about (7-13):1 (w:w), or about (11-12.7):1 (w:w); and / or relative to the total amount of the SN-38, the cholesterol, and the albumin in the composition, the SN-38 content is about 3% to about 15%, about 4% to about 15%, about 6% to about 10%, or about 8% to about 12%, or about 4% to about 10%, about 4.5% to about 9.5%, about 5% to about 9%, or about 7.5% to about 8% w / w; and / or the cholesterol content is about 5% to about 25% w / w, about 6% to about 22% w / w, about 15% to about 20% w / w, or about 4% to about 12.5% w / w, about 4.5% to about 12% w / w, about 7% to about 10% w / w, or about 7.5% to about 8% w / w; and / or the albumin content is about 64% to about 90% w / w, about 70% to about 90% w / w, or about 78% to about 93% w / w, about 79% to about 91.5% w / w, about 80% to about 90% w / w, about 82% to about 89% w / w, about 84% to about 88% w / w, or about 84.5% to about 87.5% w / w; and / or 8. The composition of claim 7, wherein the SN-38 present in the nanoparticles comprises at least about 3 wt% of the total amount of SN-38, cholesterol, and albumin in the composition, e.g., about 3 wt% to about 13 wt%, about 4 wt% to about 12 wt%, about 4 wt%, about 5 wt%, about 6 wt%, about 7 wt%, about 8 wt%, about 9 wt%, about 10 wt%, or about 11 wt%.

9. Cholesterol:SN-38 is about (1-3):1(w:w), about (1.2-2.5):1(w:w), about (1.4-2):1(w:w), about (1.5-2):1(w:w), about (1.3-1.8):1(w:w), about (1.4-1.6):1(w:w), about (1.5-1.7):1(w:w), about (1.2-1.5):1(w:w), about 1:1(w:w), about (1.4-1.5):1(w:w), or about (0.8-1.8):1(w:w), about (0.9-1.7):1(w:w), or about (1-1.4):1; albumin:SN-38 is about (5-15):1 (w:w), about (5-12):1 (w:w), about (6-12):1 (w:w), or about (7-12):1 (w:w), about (9-11):1 (w:w), about (10-12):1 (w:w), about 11:1 (w:w), or about (9-21):1 (w:w), about (9-20):1 (w:w), about (11-18):1 (w:w), or about (11.1-17.3):1 (w:w); albumin:cholesterol is about (3-10):1 (w:w), about (4-8):1 (w:w), about (5-7):1 (w:w), or about (6-21):1 (w:w), about (6.7-13):1 (w:w), about (7-13):1 (w:w), or about (11-12.7):1 (w:w); The composition of claim 7.

10. cholesterol:SN-38 is about (1-5):1 (w:w), about (1-4.5):1 (w:w), about (1-4):1 (w:w), about (1.2-3.8):1 (w:w), about (1.4-3.6):1 (w:w), about (1.6-3.4):1 (w:w), about (1.8-3.2):1 (w:w), about (2-3):1 (w:w), about (2.2-2.8):1 (w:w), about (2.4-2.6):1 (w:w), about 2.5:1 (w:w), about 1:1 (w:w), or about (0.8-1.8):1 (w:w), about (0.9-1.7):1 (w:w), or about (1-1.4):1; and / or albumin:SN-38 is about (5-25):1 (w:w), about (5-20):1 (w:w), about (6-19):1 (w:w), about (7-18):1 (w:w), about (8-16):1 (w:w), about (9-14):1 (w:w), or about (10-12):1 (w:w), or about (9-21):1 (w:w), about (9-20):1 (w:w), about (11-18):1 (w:w), or about (11.1-17.3):1 (w:w); and / or albumin:cholesterol of about (5-25):1(w:w), about (6-20):1(w:w), about (7-18):1(w:w), about (8-16):1(w:w), about (9-14):1(w:w), about (10-12):1(w:w), or about (6-21):1(w:w), about (6.7-13):1(w:w), about (7-13):1(w:w), or about (11-12.7):1(w:w); and / or Span 20:SN-38 is about (5-40):100(w:w), about (6-30):100(w:w), about (7-25):100(w:w), about (8-20):100(w:w), about (9-15):100(w:w), about (10-12):100(w:w), or about (5-10):100(w:w), about (5-9):100(w:w), about (6-8.6):100(w:w), about (6-8):100(w:w), or about (6.5-7):100(w:w); and / or relative to the total amount of the SN-38, the cholesterol, and the albumin in the composition, the SN-38 content is about 3% to about 10%, about 3.5% to about 9.5%, about 4%, about 4.5%, about 5%, about 5.5%, about 6%, about 6.5%, about 7%, about 7.5%, about 8%, about 8.5%, about 9%, or about 4% to about 10%, about 4.5% to about 9.5%, about 5% to about 9%, or about 7.5% to about 8% w / w; and / or The cholesterol content is about 4 w / w% to about 18 w / w%, about 4.5 w / w% to about 17.5 w / w%, about 5 w / w%, about 5.5 w / w%, about 6 w / w%, about 6.5 w / w%, about 7 w / w%, about 7.5 w / w%, about 8 w / w%, 8.5 w / w%, about 9 w / w%, about 9.5 w / w%, about 10 w / w%, about 10.5 w / w%, about 11 w / w%, about 11.5 w / w%, about 12 w / w%, about 12.5 w / w%, about 13 w / w%, about 13.5 w / w%, about 14 w / w%, about 14.5 w / w%, about 15 w / w%, about 15.5 w / w%, about 16 w / w%, about 16.5 w / w%, about 17 w / w%, or about 4 w / w% to about 12.5 w / w%, about 4.5 w / w% to about 12 w / w%, about 7 w / w% to about 10 w / w%, or about 7.5 w / w% to about 8 w / w%; and / or 8. The composition of claim 7, wherein the albumin content is about 78 w / w% to about 92 w / w%, about 79 w / w%, about 80 w / w%, about 81 w / w%, about 82 w / w%, about 83 w / w%, about 84 w / w%, about 85 w / w%, about 86 w / w%, about 87 w / w%, about 88 w / w%, about 89 w / w%, about 90 w / w%, about 91 w / w%, or about 78 w / w% to about 93 w / w%, about 79 w / w% to about 91.5 w / w%, about 80 w / w% to about 90 w / w%, about 82 w / w% to about 89 w / w%, about 84 w / w% to about 88 w / w%, or about 84.5 w / w% to about 87.5 w / w%.

11. the SN-38 present in the nanoparticles comprises at least about 6% to about 12% by weight, e.g., about 7% to about 11%, about 8% to about 10%, about 8.3%, or about 9% by weight of the total amount of the SN-38, the cholesterol, and the albumin in the composition; and / or 10. The composition of claim 9, wherein the SN-38 present in the nanoparticles comprises about 95% to about 99% w / w of the total amount of SN-38 in the composition, e.g., about 96% to about 99% w / w, about 97% to about 99% w / w, about 98% to about 99% w / w, or about 99% or more w / w.

12. the composition is in liquid, semi-solid, or solid form; Optionally, the solid form is in powder form, preferably a lyophilized powder; More preferably, the composition according to claim 1, characterized in that said SN-38 is present in amorphous and / or nanocrystalline form in said solid form of said composition.

13. the composition does not contain an additional stabilizer; or Preferably, the composition further comprises an additional stabilizer, such as a lyophilization stabilizer, present in an amount such that when the composition is reconstituted to form an aqueous composition (including solutions and emulsions), the content of the additional stabilizer is at least about 2% w / v, such as at least about 3% w / v, such as at least about 5% w / v, about 5% w / v to about 30% w / v, about 10% w / v to about 25% w / v, or about 15% w / v to about 20% w / v; and / or Relative to the total amount of the composition, The content of the additional stabilizer is about 60 w / w% to about 98 w / w%, for example, about 65 w / w% to about 97 w / w%, about 68 w / w% to about 96 w / w%, about 69 w / w% to about 95 w / w%, about 70 w / w% to about 94 w / w%, about 71 w / w% to about 93 w / w%, about 72 w / w% to about 92 w / w%, about 73 w / w%, about 74 w / w% %, about 75 w / w%, about 76 w / w%, about 77 w / w%, about 78 w / w%, about 79 w / w%, about 80 w / w%, about 81 w / w%, about 82 w / w%, about 83 w / w%, about 84 w / w%, about 85 w / w%, about 86 w / w%, about 87 w / w%, about 88 w / w%, about 89 w / w%, about 90 w / w%, or about 91 w / w%; For example, the content of the additional stabilizer is about 70 w / w% to about 96 w / w%, about 70 w / w% to about 90 w / w%, about 72 w / w% to about 89 w / w%, about 74 w / w% to about 88 w / w%, about 76 w / w% to about 87 w / w%, or about 80 w / w% to about 96 w / w%, about 80 w / w% to about 86 w / w%, about 81 w / w% to about 86 w / w%, about 82 w / w% to about 85 w / w%, about 83 w / w% to about 84 w / w%, or about 84 w / w% to about 95 w / w%; and / or 2. The composition of claim 1, wherein the additional stabilizer is selected from albumin (e.g., human serum albumin, recombinant human albumin, bovine serum albumin, and skim milk powder), monosaccharides, disaccharides, polysaccharides, mannitol, and any combination thereof, preferably mannitol, lactose, maltose, trehalose, dextran, glucose, and sucrose, and any composition thereof, preferably sucrose.

14. and / or, the ring-opened SN-38 in the composition comprises about 2% w / w or less, e.g., about 1.8% w / w or less, of the total amount of SN-38; and / or 2. The composition of claim 1, wherein albumin multimers are absent or substantially absent from the composition; e.g., the monomeric form of the albumin in the composition accounts for at least about 95% w / w of the total amount of albumin, e.g., at least about 96%, at least about 98%, at least about 99%, at least about 99.2%, at least about 99.4%, or at least about 99.5%.

15. The albumin is selected from human serum albumin (HSA), recombinant human albumin (rHA), bovine serum albumin, and porcine serum albumin; for example, the albumin comprises the amino acid sequence set forth in SEQ ID NO: 1; 2. The composition according to claim 1, characterized in that the albumin is preferably selected from human serum albumin (HSA) and recombinant human albumin (rHA).

16. 10. A method for preparing the composition of claim 1, comprising: (1) dissolving SN-38, lipid, and Span 20 in an organic solvent to form an organic phase; and preparing an aqueous solution of albumin as an aqueous phase; (2) mixing the organic phase and the aqueous phase to form an emulsion, the emulsion comprising nanoparticles in which the albumin encapsulates at least a portion of the SN-38 and optionally at least a portion of the lipid; and (3) removing the organic solvent from the emulsion to obtain a product containing the nanoparticles; Including; Preferably, (1) The SN-38, the lipid, and the Span 20 were dissolved in DMSO and C １～３ a first organic solvent selected from alcohols (including methanol, ethanol, and isopropanol, and any combination thereof, preferably ethanol (EtOH)); and CHCl ３ and CH ２ Cl ２ and CHCl ３ and a second organic solvent selected from a mixture of the second organic solvent with DMSO or C １～３ dissolving the albumin in a mixed organic solvent having a volume ratio of about 1:20 (v / v) to about 20:1 (v / v), for example, about 1:5 to about 5:1 (v / v), about 1:2 to about 4:1 (v / v), about 1:1 to about 4:1 (v / v), about 1.5:1 (v / v) to about 3:1 (v / v), or about 2:1 (v / v) to 7:3 (v / v) to form an organic phase; preparing an aqueous solution of the albumin as an aqueous phase; (2) mixing the organic phase and the aqueous phase to form an emulsion, the emulsion comprising the nanoparticles, wherein the albumin encapsulates at least a portion of the SN-38 and optionally at least a portion of the lipid; (3) removing the organic solvent; and (4) Optionally, sterilizing the product obtained in step (3) by filtering it, preferably through a filter membrane of about 0.2 μm. Including; Optionally, the second organic solvent is CHCl ３ , or CH ２ Cl ２ and CHCl ３ and optionally, CH ２ Cl ２ vs. CHCl ３ 2. A method for preparing a composition according to claim 1, characterized in that the volume ratio of is about 2:5 to 1:1, preferably about 2:

5.

17. 17. The method of claim 16, wherein in step (2), the organic phase:aqueous phase ratio is from about 1:2 (v / v) to about 1:50 (v / v), for example, from about 1:5 (v / v) to about 1:20 (v / v), from about 1:7 (v / v) to about 1:15 (v / v), from about 1:10 (v / v) to about 1:12 (v / v), for example, from about 1:5 (v / v) to about 1:12 (v / v), from about 1:5 (v / v) to about 1:12 (v / v), about 1:6 (v / v), about 1:7 (v / v), or about 1:10 (v / v).

18. A pharmaceutical composition comprising the composition of any one of claims 1 to 15, and optionally a pharmaceutically acceptable carrier.

19. 16. A medicament for treating an SN-38-sensitive tumor in a subject, comprising the composition of any one of claims 1 to 15, wherein the tumor is preferably selected from colorectal cancer, small cell lung cancer, lymphatic cancer, breast cancer (preferably triple-negative breast cancer), esophageal cancer, gastric cancer, liver cancer, renal cancer, pancreatic cancer, uterine cancer, and ovarian cancer.

20. 1. A method for preparing a composition with improved properties, the composition comprising SN-38, a lipid, and an albumin, the albumin encapsulating at least a portion of the SN-38 and optionally at least a portion of the lipid to form nanoparticles; The method is characterized in that Span 20 is added during the preparation of the composition; Optionally, the composition does not contain an additional stabilizer; and / or Optionally, the improved properties include improved stability; for example, when the composition is in liquid form, the improved stability includes a reduction in the formation or content of albumin multimers (e.g., the albumin multimers are absent or substantially absent from the composition, or the albumin multimers account for at most 5% w / w of the total amount of albumin, e.g., at most about 4%, at most about 2%, at most about 1.5%, at most about 1.2%, at most about 1.1%, at most about 1%, or at most about 0.8%), and / or a reduction in particle size of the nanoparticles during preparation, storage, and / or use of the composition; and / or Optionally, the composition is as defined in any one of claims 1 to 15; Preferably, the method comprises: (1) dissolving the SN-38, the lipid, and the Span 20 in an organic solvent to form an organic phase; and preparing an aqueous solution of the albumin as an aqueous phase; (2) mixing the organic phase and the aqueous phase to form an emulsion, the emulsion comprising the nanoparticles, wherein the albumin encapsulates at least a portion of the SN-38 and optionally at least a portion of the lipid; and (3) removing the organic solvent from the emulsion to obtain a product containing the nanoparticles; Including; More preferably, the method comprises: (1) The SN-38, the lipid, and the Span 20 were dissolved in DMSO and C １～３ a first organic solvent selected from alcohols (including methanol, ethanol, and isopropanol, and any combination thereof, preferably ethanol (EtOH)); and CHCl ３ and CH ２ Cl ２ and CHCl ３ and a second organic solvent selected from a mixture of the second organic solvent with DMSO or C １～３ dissolving the albumin in a mixed organic solvent having a volume ratio of about 1:20 (v / v) to about 20:1 (v / v), for example, about 1:5 to about 5:1 (v / v), about 1:2 to about 4:1 (v / v), about 1:1 to about 4:1 (v / v), about 1.5:1 (v / v) to about 3:1 (v / v), or about 2:1 (v / v) to 7:3 (v / v) to form an organic phase; preparing an aqueous solution of the albumin as an aqueous phase; (2) mixing the organic phase and the aqueous phase to form an emulsion, the emulsion comprising the nanoparticles, wherein the albumin encapsulates at least a portion of the SN-38 and optionally at least a portion of the lipid; (3) removing the organic solvent; and (4) Optionally, sterilizing the product obtained in step (3) by filtering it, preferably through a filter membrane of about 0.2 μm. Including; Optionally, the second organic solvent is CHCl ３ , or CH ２ Cl ２ and CHCl ３ and optionally, CH ２ Cl ２ vs. CHCl ３ The volume ratio of is about 2:5 to 1:1, preferably about 2:5; Methods for preparing compositions with improved properties.

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