Subcutaneous administration of nanoparticles comprising mTOR inhibitor and albumin to treat disease

Abstract

The name of the invention is subcutaneous administration of nanoparticles comprising an mTOR inhibitor and albumin to treat disease. The present invention provides compositions and devices for subcutaneous administration of compositions comprising nanoparticles comprising an mTOR inhibitor and albumin. The present application also provides methods of treating a disease by subcutaneously administering to an individual a composition comprising nanoparticles comprising an mTOR inhibitor and albumin.

Description

[0001] This application is a divisional application. The filing date of the original application is March 18, 2020, the application number is 2020800345439 (PCT / US2020 / 023366), and the invention title is "Subcutaneous Administration of Nanoparticles Comprising an mTOR Inhibitor and Albumin for Treating Diseases".

[0002] Cross - reference to related applications

[0003] This application claims the priority benefit of U.S. Provisional Application No. 62 / 820,842, entitled "Subcutaneous Administration of Nanoparticles Comprising an mTOR Inhibitor and Albumin for Treating Diseases", filed on March 19, 2019; and U.S. Provisional Application No. 62 / 820,838, entitled "Methods and Compositions for Treating Pulmonary Hypertension", filed on March 19, 2019; each of which is incorporated herein by reference for all purposes. Technical field

[0004] This application relates to compositions, devices, and methods for subcutaneous administration of nanoparticles comprising an mTOR inhibitor and albumin. This application further relates to methods of treating an individual, which include subcutaneous administration of a composition comprising nanoparticles comprising an mTOR inhibitor and albumin. Background art

[0005] Mammalian target of rapamycin (mTOR) is a protein kinase known to regulate various cellular processes including cell survival, proliferation, stress, and metabolism. Many inhibitors of mTOR, including rapamycin, can effectively treat various diseases including certain cancers. Many mTOR inhibitors, such as rapamycin, are known to have poor water solubility and thus require excipients such as surfactants and solvents. These excipients can cause irritation, inflammation, and reduced efficacy, especially upon parenteral administration, such as subcutaneous administration.

[0006] Accordingly, there is a need in the art for improved nanoparticle formulations comprising an mTOR inhibitor that are stable upon administration and / or do not cause unacceptable toxicological effects, such as upon subcutaneous administration. There is also a need to develop formulations of nanoparticles comprising an mTOR inhibitor that are dry, such as lyophilized, and can be more readily formulated and / or delivered. Finally, there is a need in the art to reduce the risk of mishandling the dry composition prior to administration.

[0007] The disclosures of all publications, patents, patent applications, and published patent applications mentioned herein are incorporated herein by reference in their entirety. Summary of the invention

[0008] The present application provides a method for treating a disease in an individual, which comprises subcutaneously administering to the individual a pharmaceutical composition comprising nanoparticles comprising an mTOR inhibitor and albumin, wherein the amount of the mTOR inhibitor in the pharmaceutical composition is about 0.1 mg / m 2 to about 10 mg / m 2 per administration. In some embodiments, the amount of the mTOR inhibitor in the pharmaceutical composition is about 1 mg / m 2 to about 10 mg / m 2 per administration. In some embodiments, the amount of the mTOR inhibitor in the pharmaceutical composition is about 5 mg / m 2 per administration.

[0009] In some embodiments according to any of the methods described herein, the pharmaceutical composition further comprises a sugar.

[0010] In some embodiments according to any of the methods described herein, the pharmaceutical composition is administered once a week or less. In some embodiments, the pharmaceutical composition is administered once a week. In some embodiments, the pharmaceutical composition is administered twice every three weeks.

[0011] In some embodiments according to any of the methods described herein, the disease is cancer. In some embodiments according to any of the methods described herein, the disease is a mitochondrial disease.

[0012] In some embodiments according to any of the methods described herein, the individual is a human.

[0013] The present application also provides a method for delivering an effective amount of an mTOR inhibitor to a target tissue of an individual, which comprises subcutaneously administering a pharmaceutical composition comprising nanoparticles comprising an mTOR inhibitor and albumin. In some embodiments, the mTOR inhibitor is rapamycin. In some embodiments, the pharmaceutical composition further comprises a sugar. In some embodiments, the pharmaceutical composition is at a dose of about 0.1 mg / m 2 to about 10 mg / m 2 per administration. In some embodiments, the target tissue is the brain tissue of the individual.

[0014] In some embodiments according to any of the methods described herein, the average diameter of the nanoparticles in the pharmaceutical composition is not greater than about 120 nm. In some embodiments according to any of the methods described herein, the nanoparticles comprise an mTOR inhibitor coated with albumin. In some embodiments according to any of the methods described herein, the albumin is human albumin. In some embodiments according to any of the methods described herein, the mTOR inhibitor is the drug limus. In some embodiments according to any of the methods described herein, the mTOR inhibitor is rapamycin.

[0015] The present application also provides a pharmaceutical composition suitable for subcutaneous administration to an individual, comprising: a) nanoparticles comprising an mTOR inhibitor and albumin, and b) a sugar. In some embodiments, the sugar is selected from alginate, starch, lactose, pullulan, hyaluronic acid, chitosan, glucose, galactose, mannose, N-acetylglucosamine, sucrose, N-acetyl-D-galactosamine, maltose or trehalose. In some embodiments, the sugar is sucrose. In some embodiments, the sugar is trehalose. In some embodiments, the concentration of the mTOR inhibitor in the pharmaceutical composition is at least about 5 mg / ml. In some embodiments, the concentration of the mTOR inhibitor in the pharmaceutical composition is at least about 50 mg / ml. In some embodiments, the average diameter of the nanoparticles in the pharmaceutical composition is not greater than about 120 nm. In some embodiments, the nanoparticles in the pharmaceutical composition comprise an mTOR inhibitor coated with albumin. In some embodiments, the albumin in the pharmaceutical composition is human albumin. In some embodiments, the mTOR inhibitor in the pharmaceutical composition is a limus drug. In some embodiments, the mTOR inhibitor is rapamycin.

[0016] The present application also provides a device for subcutaneous administration to an individual of a pharmaceutical composition comprising nanoparticles comprising an mTOR inhibitor and albumin, the device comprising a) a drug chamber containing the pharmaceutical composition in dry form and a solution chamber containing a reconstitution solution; and b) a removable partition separating the drug chamber and the solution chamber, wherein removal of the partition causes mixing of the dry pharmaceutical composition and the reconstitution solution to form a reconstituted pharmaceutical composition. In some embodiments, the device is a syringe, the syringe comprising a needle fixed to the end of the syringe and a plunger capable of expelling the reconstituted pharmaceutical composition from the syringe. In some embodiments, the pharmaceutical composition of the device further comprises a sugar. In some embodiments of the device, the mTOR inhibitor is a limus drug. In some embodiments of the device, the mTOR inhibitor is rapamycin.

[0017] The present application also provides a kit comprising any of the devices described herein for treating a disease. In some embodiments, the kit further comprises instructions for using the kit to treat cancer. In some embodiments, the kit further comprises instructions for using the kit to treat mitochondrial diseases. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 Shows the rapamycin concentration in whole blood samples taken from rats after subcutaneous (SC) or intravenous (IV) administration of a single dose of nab-rapamycin (ABI-009) from 0 to 24 hours after administration.

[0019] Figure 2Shows the rapamycin concentration in whole blood samples taken from rats after single-dose subcutaneous (SC) or intravenous (IV) administration of nab-rapamycin (ABI-009) at 0 to 168 hours post-administration.

[0020] Figure 3 Shows the rapamycin concentration in whole blood samples taken from rats after single-dose subcutaneous (SC) or intravenous (IV) administration of nab-rapamycin (ABI-009) at 0 to 24 hours post-administration.

[0021] Figure 4 Shows the bioavailability of single-dose nab-rapamycin (ABI-009) after subcutaneous (subQ) or intravenous (IV) administration in rats, as indicated by the calculated area under the curve (AUC).

[0022] Figure 5 Shows the rapamycin concentration in rat bone marrow (upper panel) or brain (lower panel) at 24 or 168 hours after single-dose subcutaneous (subQ) or intravenous (IV) administration of nab-rapamycin (ABI-009).

[0023] Figure 6 Shows the rapamycin concentration in rat heart (upper panel) or liver (lower panel) at 24 or 168 hours after single-dose subcutaneous (subQ) or intravenous (IV) administration of nab-rapamycin (ABI-009).

[0024] Figure 7 Shows the rapamycin concentration in rat lung (upper panel) or pancreas (lower panel) at 24 or 168 hours after single-dose subcutaneous (subQ) or intravenous (IV) administration of nab-rapamycin (ABI-009).

[0025] Figure 8 Shows the comparison of rapamycin concentration over time in the brain or whole blood of rats at 24, 72, and 120 hours after single subcutaneous administration of nab-rapamycin (ABI-009) at a dose of 1.7 mg / kg, 9.5 mg / kg, or 17 mg / kg.

[0026] Figure 9 Shows the comparison of histopathological scores of skin evaluations in rats from different treatment groups.

[0027] Figure 10 Is a representative histogram of rat skin from Group 1 (0.9% saline). Histological lesions were limited to the aggregation of mixed inflammatory cells (black arrows) within the subcutaneous tissue (SC). The dermis (D) and epidermis (E) are indicated.

[0028] Figure 11Is a representative histogram of rat skin from Group 2 (HSA in 0.9% saline). Multifocal mixed inflammatory cell aggregates (black arrows) can be seen within the subcutaneous (SC). The epidermis (E) and dermis (D) are not significant.

[0029] Figure 12 Is a representative histogram of rat skin from Group 3 (ABI-009, 1.7 mg / kg). Minimal mixed inflammatory cell infiltration (black arrow) can be seen in the subcutaneous tissue (SC). The epidermis (E) and dermis (D) are indicated.

[0030] Figure 13 Is a representative histogram of rat skin from Group 4 (ABI-009, 5 mg / kg). Dispersed mixed inflammatory cell infiltration (right arrow) and minimal necrotic sites (left arrow) are present in the subcutaneous (SC). The epidermis (E) and dermis (D) are not significant.

[0031] Figure 14 Is a representative histogram of rat skin from Group 4 (ABI-009, 10 mg / kg). Captured subcutaneous (SC) mixed inflammatory cell infiltration (right arrow) and necrotic areas (left arrow). The epidermis (E) and dermis (D) are not significant.

[0032] Figure 15 Shows the mean rapamycin levels in the blood of rats administered ABI-009 at 1.7 mg / kg, 5 mg / kg, or 10 mg / kg.

[0033] Figure 16 Shows the tumor growth results of a human hepatocellular carcinoma mouse xenograft model after treatment with saline (Group 1), ABI-009 (intravenous route; Group 2), rapamune (oral administration; Group 3), and ABI-009 (subcutaneous route; Group 4) for 0 - 15 days.

[0034] Figure 17 Shows the changes in the body weight of mice in a human hepatocellular carcinoma mouse xenograft model after treatment with saline (Group 1), ABI-009 (intravenous route; Group 2), rapamune (oral administration; Group 3), and ABI-009 (subcutaneous route; Group 4) for 0 - 15 days. Detailed implementation

[0035] The present disclosure provides methods for subcutaneous administration of a composition as described herein, such as a composition comprising nanoparticles comprising an mTOR inhibitor (such as rapamycin) and albumin. In another aspect, the present disclosure provides methods for delivering an effective amount of an mTOR inhibitor (such as rapamycin) to a target tissue, such as brain, bone marrow, heart, liver, lung, or pancreatic tissue, by subcutaneous administration of a composition comprising nanoparticles comprising an mTOR inhibitor (such as rapamycin) and albumin. In another aspect, the present disclosure provides methods for maintaining blood levels of an mTOR inhibitor (such as rapamycin), comprising subcutaneous administration of a composition comprising nanoparticles comprising an mTOR inhibitor (such as rapamycin) and albumin.

[0036] The present disclosure also provides methods for treating a disease, comprising subcutaneous administration of a composition comprising nanoparticles comprising an mTOR inhibitor (such as rapamycin) and albumin. In some embodiments, the disease is cancer. In some embodiments, individuals with cancer are selected for treatment based on having an aberration in mTOR activation. In some embodiments, the disease is a mitochondrial disorder.

[0037] The present disclosure also provides compositions suitable for subcutaneous administration to an individual, including pharmaceutical compositions, which comprise nanoparticles comprising an mTOR inhibitor (such as rapamycin) and albumin, and methods of administering such compositions. In one aspect, the composition may comprise one or more reagents for enhancing dissolution of the composition in dry form and / or enhancing the stability of the composition. The additional one or more reagents may comprise sugars. The amount of sugar present may effectively increase solubility, such as the dissolution rate after addition of an aqueous solution to the composition in dry form, and / or promote the stability of the composition.

[0038] In another aspect, the present disclosure provides a device for subcutaneous administration of a pharmaceutical composition as described herein. The device comprises a drug chamber containing the pharmaceutical composition in dry form and a solution chamber containing a reconstitution solution. The device further comprises a removable partition separating the drug chamber and the solution chamber, wherein removal or actuation of the partition causes or permits mixing of the dry pharmaceutical composition and the reconstitution solution, thereby forming a reconstituted pharmaceutical composition. The device may be a syringe, which may further comprise a plunger. After reconstitution of the composition, the device is capable or adaptable to be able to administer the composition subcutaneously to an individual. The present disclosure also provides methods of subcutaneous administration of a composition comprising nanoparticles comprising an mTOR inhibitor and albumin using a device as described herein.

[0039] Definitions

[0040] It is to be understood that aspects and embodiments of the invention described herein include “consisting of” and / or “consisting essentially of” the aspects and embodiments.

[0041] As used herein, albumin can be "associated" with an mTOR inhibitor (such as rapamycin). For example, a composition includes an mTOR inhibitor associated with albumin. "Association" or "associated" is used herein in a general sense and refers to albumin that affects the behavior and / or properties of an mTOR inhibitor (such as rapamycin) in an aqueous composition. For example, if albumin makes an mTOR inhibitor (such as rapamycin) more readily suspendable in an aqueous medium compared to a composition without albumin, then the albumin and the mTOR inhibitor (such as rapamycin) are considered to be "associated". As another example, if albumin stabilizes an mTOR inhibitor (such as rapamycin) in an aqueous suspension, then the albumin and the mTOR inhibitor (such as rapamycin) are associated. For example, albumin and an mTOR inhibitor can be present in a particle or nanoparticle, which is further described herein.

[0042] General reference to "composition" can include any pharmaceutical composition described herein.

[0043] The term "effective amount" as used herein refers to an amount of a compound or composition sufficient to treat a particular disorder, condition or disease such as to ameliorate, mitigate, relieve and / or delay one or more of its symptoms. As understood in the art, an "effective amount" can be one or more doses, i.e., a single dose or multiple doses may be required to achieve the desired therapeutic endpoint. An effective amount can be considered in the context of administering one or more therapeutic agents, and if administered in combination with one or more agents, an effective amount of a nanoparticle composition (e.g., a composition comprising rapamycin and albumin) may achieve or have achieved a desirable or beneficial result.

[0044] As used herein, represents nanoparticle albumin binding, and "nab-rapamycin" is an albumin-stabilized rapamycin nanoparticle formulation. Nab-rapamycin is also known as nab-sirolimus and has been previously described. See, for example, WO2008 / 109163A1, WO2014 / 151853, WO2008 / 137148A2 and WO2012 / 149451A1, each of which is incorporated herein by reference in its entirety.

[0045] As used herein, "pharmaceutically acceptable" or "pharmacologically compatible" refers to a material that is not biologically or otherwise undesirable, e.g., the material can be incorporated into a pharmaceutical composition administered to a patient without causing any significant undesirable biological effects or interactions in a harmful manner with any other component of the composition in which it is contained. Pharmaceutically acceptable carriers or excipients preferably meet the required standards of toxicological and production testing and / or are included in the Inactive Ingredient Guides prepared by the U.S. Food and Drug Administration.

[0046] As used herein, "treatment" or "treating" is a method of obtaining a beneficial or desired result, including a clinical result. For the purposes of the present invention, beneficial or desired clinical results include, but are not limited to, one or more of the following: alleviation of one or more symptoms caused by a disease, reduction in the severity of a disease, stabilization of a disease (e.g., prevention or delay of disease progression), prevention or delay of the spread of a disease (e.g., metastasis), prevention or delay of the recurrence of a disease, reduction in the recurrence rate of a disease, delay or slowing of the progression of a disease, improvement in the state of a disease, provision of remission (partial or complete) of a disease, reduction in the dosage of one or more other drugs required to treat a disease, delay in the progression of a disease, improvement in quality of life, and / or extension of survival. In some embodiments, treatment reduces the severity of one or more cancer-related symptoms by at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or 100% as compared to the corresponding symptoms of the same subject prior to treatment or as compared to the corresponding symptoms of other subjects not receiving treatment. "Treatment" also includes reduction of the pathological consequences of cancer. The methods of the present invention contemplate any one or more of these aspects of treatment.

[0047] The terms "recurrence", "relapse", or "relapsed" refer to the recurrence of cancer or a disease after clinical assessment of the disappearance of the disease. The diagnosis of distant metastasis or local recurrence may be considered a recurrence.

[0048] The terms "refractory" or "resistant" refer to a cancer or disease that does not respond to treatment.

[0049] It should be understood that the embodiments of the invention described herein include "consisting of" and / or "consisting essentially of" embodiments.

[0050] The mention herein of "about" a value or parameter includes (and describes) variations that are directed to that value or parameter per se. For example, the description of "about X" includes the description of "X".

[0051] As used herein, reference to "not" a value or parameter generally means and describes "different from" a value or parameter. For example, the method is not for treating cancer type X means the method is for treating a cancer type other than X.

[0052] As used in this specification and the appended claims, the singular forms "a", "an", and "the" include plural referents unless the context clearly dictates otherwise.

[0053] Method of subcutaneous administration

[0054] Provided herein are methods of subcutaneous administration of a composition, such as a pharmaceutical composition, that includes an mTOR inhibitor, such as rapamycin, and albumin.

[0055] In some embodiments, provided is a method for delivering an effective amount of an mTOR inhibitor (such as rapamycin) to a target tissue of an individual, such as brain, bone marrow, heart, liver, lung, or pancreatic tissue, the method comprising subcutaneous administration of a composition, such as a pharmaceutical composition, comprising nanoparticles that include an mTOR inhibitor (such as rapamycin) and albumin. In some embodiments, the individual has a tumor in a target tissue, such as the brain.

[0056] In some embodiments, provided is a method for delivering an effective amount of an mTOR inhibitor (such as rapamycin) to the brain of an individual, the method comprising subcutaneous administration of a composition, such as a pharmaceutical composition, comprising nanoparticles that include an mTOR inhibitor (such as rapamycin) and albumin, wherein the dose of the mTOR inhibitor (such as rapamycin) in the nanoparticles for delivering an effective amount of the mTOR inhibitor (such as rapamycin) to the brain is from about 0.1 mg / m 2 to about 10 mg / m 2 of any value, and values and ranges therein.

[0057] In some embodiments, the method comprises maintaining a blood level of an mTOR inhibitor (such as rapamycin) in an individual, the method comprising subcutaneous administration of a composition, such as a pharmaceutical composition, comprising nanoparticles that include an mTOR inhibitor (such as rapamycin) and albumin. In some embodiments, the blood level of the mTOR inhibitor (such as rapamycin) is at least any one of 1 ng / ml, 5 ng / ml, 10 ng / ml, 25 ng / ml, 50 ng / ml, 75 ng / ml, 100 ng / ml, 150 ng / ml, or 200 ng / ml, and values and ranges therein. In some embodiments, the individual has a tumor.

[0058] In some embodiments, provided herein are methods of treating a disease in an individual, including subcutaneously administering to the individual a pharmaceutical composition comprising nanoparticles comprising an mTOR inhibitor and albumin. In some embodiments, the amount of the mTOR inhibitor in the pharmaceutical composition is about 0.1 mg / m 2 to about 10 mg / m 2 , such as about 1 mg / m 2 to about 10 mg / m 2 per dose. In an exemplary non-limiting embodiment, the amount of the mTOR inhibitor in the pharmaceutical composition is about 5 mg / m 2 per dose.

[0059] In some embodiments, the amount of the mTOR inhibitor in the composition is below the level that induces toxicological effects (e.g., effects above the clinically acceptable toxicity level) or is at a level of potential side effects that can be controlled or tolerated when the mTOR inhibitor nanoparticle composition is subcutaneously administered to the individual. In some embodiments, the toxicological effect is a rash associated with the subcutaneous administration of the pharmaceutical composition.

[0060] In some embodiments, the concentration of the mTOR inhibitor (such as rapamycin) in the mTOR inhibitor nanoparticle composition is between about 0.1 mg / ml and about 100 mg / ml, including, for example, any one of about 0.1 mg / ml to about 50 mg / ml, about 0.1 mg / ml to about 50 mg / ml, about 0.1 mg / ml to about 40 mg / ml, about 0.1 mg / ml to about 10 mg / ml, or about 0.1 mg / ml to about 5 mg / ml, about 5 mg / ml to about 100 mg / ml, about 5 mg / ml to about 50 mg / ml, about 5 mg / ml to about 40 mg / ml, about 7.5 mg / ml to about 100 mg / ml, about 7.5 mg / ml to about 50 mg / ml, about 7.5 mg / ml to about 40 mg / ml, and values and ranges therein. In some embodiments, the concentration of the mTOR inhibitor (such as rapamycin) in the mTOR inhibitor nanoparticle composition is at least any one of 5 mg / ml, 7.5 mg / ml, 10 mg / ml, or 20 mg / ml.

[0061] In some embodiments, the effective amount of the mTOR inhibitor (such as rapamycin) in the mTOR inhibitor nanoparticle composition is in any one of the following ranges: about 0.1 mg / m 2 to about 5 mg / m 2 , about 5 mg / m 2 to about 10 mg / m 2 , about 10 mg / m 2 to about 20 mg / m 2, from about 10 to about 30 mg / m 2 , from about 10 to about 45 mg / m 2 , from about 10 to about 60 mg / m 2 , from about 20 to about 30 mg / m 2 , from about 20 to about 45 mg / m 2 , from about 20 to about 60 mg / m 2 , from about 30 to about 45 mg / m 2 , from about 30 to about 60 mg / m 2 , or from about 45 to about 60 mg / m 2 , each including the endpoints. In an exemplary non-limiting embodiment, the effective amount of the mTOR inhibitor (such as rapamycin) in the mTOR inhibitor nanoparticle composition is between about 0.1 mg / m 2 and about 10 mg / m 2 . In another exemplary non-limiting embodiment, the effective amount of the mTOR inhibitor (such as rapamycin) in the mTOR inhibitor nanoparticle composition is between about 1 mg / m 2 and 10 mg / m 2 , such as 5 mg / m 2 .

[0062] In some embodiments, the dosing frequency of administration of the mTOR inhibitor nanoparticle composition (such as the rapamycin / albumin nanoparticle composition) includes but is not limited to once a day, once every two days, once every three days, once every four days, once every five days, once every six days, once a week without interruption, three times every four weeks (such as on days 1, 8, and 15 of a 28-day cycle), once every three weeks, once every two weeks, or twice every three weeks. In some embodiments, the mTOR inhibitor nanoparticle composition (such as the rapamycin / albumin nanoparticle composition) is administered once every about 2 weeks, once every 3 weeks, once every 4 weeks, once every 6 weeks, or once every 8 weeks. In some embodiments, the mTOR inhibitor nanoparticle composition (such as the rapamycin / albumin nanoparticle composition) is administered at least any one of about 1, 2, 3, 4, 5, 6, or 7 times per week (i.e., once a day). In some embodiments, the interval between each administration is less than any one of about 6 months, 3 months, 1 month, 20 days, 15 days, 14 days, 13 days, 12 days, 11 days, 10 days, 9 days, 8 days, 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, or 1 day. In some embodiments, the interval between each administration is greater than any one of about 1 week, 2 weeks, 3 weeks, 4 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 8 months, or 12 months. In some embodiments, the dosing regimen is without interruption. In some embodiments, the interval between each administration is not greater than about one week.

[0063] Administration of the mTOR inhibitor nanoparticle composition (such as rapamycin / albumin nanoparticle composition) can be extended for an extended period of time, such as from about 1 month to about 7 years. In some embodiments, the mTOR inhibitor nanoparticle composition (such as rapamycin / albumin nanoparticle composition) is administered for a period of at least about any of 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 18, 24, 30, 36, 48, 60, 72, or 84 months.

[0064] The auxiliary agents and adjuvants in any of the compositions may include, for example, preservatives, wetting agents, suspending agents, flavoring agents, emulsifiers and dispersants. Prevention of microbial action is usually provided by various antibacterial and antifungal agents, such as parabens, chlorobutanol, phenol, sorbic acid, etc. Isotonic agents may also be included. The absorption of injectable drug forms may be prolonged by using delayed absorption agents such as aluminum monostearate and gelatin. Auxiliary agents may also include wetting agents, emulsifiers, pH buffers and antioxidants, such as citric acid, sorbitan monolaurate, triethanolamine oleate, butylated hydroxytoluene, etc.

[0065] Treatment according to any dosing regimen, such as the exemplary dosing regimen discussed above, can be repeated for multiple cycles (such as 1, 2, 3, 4, 5, 6 or more cycles, such as about 1-10 cycles, 1-7 cycles, 1-5 cycles, 1-4 cycles, 1-3 cycles). In some embodiments, treatment according to a specific dosing regimen is repeated for at least two, three or more cycles. In some embodiments, treatment according to a specific dosing regimen is repeated continuously (i.e., without intervals) for at least two, three or more cycles.

[0066] In some embodiments, there is an interval between two adjacent cycles. In some embodiments, the interval is at least about 1, 2, 3, or 4 weeks. In some embodiments, the interval is at least about 1, 2, 3, 4, 5, 6 months or longer. In some embodiments, the interval is about a period of time that allows the individual to gain weight (e.g., after the interval, the individual's weight is about or at least about 90%, 92%, 95%, 97% of the weight before starting treatment).

[0067] In some embodiments, the pharmaceutical composition is administered only once.

[0068] Subcutaneous administration device

[0069] This application demonstrates that subcutaneous administration of a drug comprising an mTOR inhibitor / albumin nanoparticle exhibits better tolerance and bioavailability properties compared to intravenous administration. In some embodiments, a composition comprising nanoparticles containing an mTOR inhibitor and albumin can be stored in a dry form, such as a lyophilized form. Preparation of the dry composition for administration requires reconstitution with an aqueous solution such as water. One aspect of this application provides a device that contains a stable, fixed-dose dry composition proximate to a reconstitution solution. The device includes a septum that, through a limited operation, allows predictable and reproducible reconstitution of the dry composition, and then subcutaneous administration of the composition using the device. The device improves handling of the dry composition by, for example, reducing the handling time of the composition, reducing the chance of user error, and ensuring reproducibility and consistency of the reconstitution process. These advantages are obtained, for example, by eliminating the step of manually adding the reconstitution solution to the dry composition and completely eliminating the step of loading the reconstituted composition into a syringe.

[0070] Accordingly, provided herein is a device for subcutaneous administration of a composition, such as a pharmaceutical composition comprising nanoparticles containing an mTOR inhibitor and albumin. The devices described herein are particularly suitable for subcutaneous administration of a composition in dry form by continuously reconstituting the composition in dry form and then administering the reconstituted composition. In one aspect, the device includes a drug chamber containing a pharmaceutical composition in dry form (such as those described herein, such as, for example, a lyophilized pharmaceutical composition) and a solution chamber containing a reconstitution solution. In another aspect, the device includes a septum separating the drug chamber and the solution chamber. Removal of the septum or actuation of the septum allows and / or causes mixing of the dry pharmaceutical composition and the reconstitution solution, thereby forming a reconstituted pharmaceutical composition. The reconstituted pharmaceutical composition may be suitable for subcutaneous administration to an individual, such as a human. In some embodiments, the device is a syringe that includes a solution chamber and a drug chamber. In some embodiments, the syringe further includes a plunger capable of expelling the reconstitution solution from the device. In some embodiments, the syringe also includes an injection needle suitable for subcutaneous administration that is fixed to the end of the syringe, such as a subcutaneous injection needle.

[0071] Further provided herein is a composition for subcutaneous administration contained within a device, wherein the composition comprises a pharmaceutical composition in lyophilized form, and wherein the device includes a solution chamber containing a reconstitution solution and a septum separating the drug chamber (containing the pharmaceutical composition) and the reconstitution solution, wherein removal or actuation of the septum allows and / or causes mixing of the dry pharmaceutical composition and the reconstitution solution, thereby forming a reconstituted pharmaceutical composition.

[0072] In some embodiments, the septum of the device may include a guard that prevents accidental removal or actuation of the septum. In some embodiments, the guard is removed from the device before removing or actuating the septum. In some embodiments, the guard is actuated to allow removal or actuation of the septum.

[0073] In some embodiments, the device is a syringe that includes a suitable needle for injection. In some embodiments, the device is a syringe adapted to be connected to a suitable needle for injection. Depressing the plunger of the syringe causes the reconstituted syringe to be expelled through the needle.

[0074] In some embodiments, there is provided a device for subcutaneous administration of a pharmaceutical composition comprising nanoparticles comprising an mTOR inhibitor (such as rapamycin) and albumin, the device comprising a drug chamber containing the pharmaceutical composition in dry form, a solution chamber containing a reconstitution solution, and a removable septum separating the drug chamber and the solution chamber, wherein removal of the septum causes mixing of the dry pharmaceutical composition and the reconstitution solution to form a reconstituted pharmaceutical composition, wherein the dose of the mTOR inhibitor (such as rapamycin) in the nanoparticles is any one of about 0.2 mg to about 100 mg, about 0.2 mg to about 10 mg, about 10 mg to about 20 mg, about 20 mg to about 30 mg, about 30 mg to about 40 mg, about 40 mg to about 50 mg, about 50 mg to about 60 mg, about 60 mg to about 70 mg, about 70 mg to about 80 mg, about 80 mg to about 90 mg, about 90 mg to about 100 mg, each including the endpoints.

[0075] In some embodiments, a device for subcutaneous administration of a pharmaceutical composition comprising nanoparticles comprising an mTOR inhibitor (such as rapamycin) and albumin is provided, the pharmaceutical composition optionally comprising a sugar, the device comprising a drug chamber containing the pharmaceutical composition in dry form, a solution chamber containing a reconstitution solution, and a removable separator separating the drug chamber and the solution chamber, wherein removal of the separator causes mixing of the dry pharmaceutical composition and the reconstitution solution to form a reconstituted pharmaceutical composition, wherein the dose of the mTOR inhibitor (such as rapamycin) in the nanoparticles is any one of from about 0.2 mg to about 100 mg, from about 0.2 mg to about 10 mg, from about 10 mg to about 20 mg, from about 20 mg to about 30 mg, from about 30 mg to about 40 mg, from about 40 mg to about 50 mg, from about 50 mg to about 60 mg, from about 60 mg to about 70 mg, from about 70 mg to about 80 mg, from about 80 mg to about 90 mg, from about 90 mg to about 100 mg, each including the endpoints. In some embodiments, the sugar is selected from alginate, starch, lactose, pullulan, hyaluronic acid, chitosan, glucose, galactose, mannose, N-acetylglucosamine, sucrose, N-acetyl-D-galactosamine, maltose, or trehalose.

[0076] Also provided herein is a method of subcutaneous administration of the composition described herein using the device described herein. In one non-limiting exemplary embodiment, the composition comprises the pharmaceutical composition in dry form, the pharmaceutical composition comprising nanoparticles comprising rapamycin and albumin. In one non-limiting exemplary embodiment, the method of subcutaneous administration comprises selecting an individual for subcutaneous administration of the pharmaceutical composition, removing or actuating the separator, waiting a specified period of time for the dry composition and the reconstitution solution to form a reconstituted pharmaceutical composition, inserting the needle into the individual at an appropriate angle, pressing down on the plunger with an appropriate force, and removing the needle from the individual.

[0077] Disease to be treated

[0078] The compositions, methods, and devices described herein can be used to treat one or more diseases in an individual such as a human. In some embodiments, the disease or diseases are one or more of pulmonary hypertension, central nervous system disorders, mitochondrial disorders, or cancer.

[0079] I. Pulmonary hypertension

[0080] Pulmonary hypertension (PH) is a syndrome characterized by elevated pulmonary artery pressure. PH is defined hemodynamically as a systolic pulmonary artery pressure greater than 30 mm Hg or an estimated mean pulmonary artery pressure greater than 25 mm Hg. See Zaiman et al., Am. J. Respir. Cell Mol. Biol. 33:425-31 (2005).

[0081] In some embodiments of any of the methods described herein, one or more diseases to be treated include pulmonary hypertension. In some embodiments, the pulmonary hypertension is any one of pulmonary arterial hypertension (PAH), idiopathic pulmonary arterial hypertension (IPAH), hereditary pulmonary arterial hypertension (HPAH), drug- and toxin-induced PAH, PAH associated with connective tissue disorders, and PAH associated with congenital heart defects.

[0082] In some embodiments, the pulmonary hypertension is severe pulmonary arterial hypertension. In some embodiments, the pulmonary hypertension is World Health Organization [WHO] functional class II, III, or IV pulmonary arterial hypertension. In some embodiments, the pulmonary hypertension is WHO functional class II pulmonary arterial hypertension. In some embodiments, the pulmonary hypertension is WHO functional class III pulmonary arterial hypertension. In some embodiments, the pulmonary hypertension is WHO functional class IV pulmonary arterial hypertension.

[0083] Central nervous system disorders

[0084] Central nervous system diseases, also known as central nervous system disorders, are a group of neurological disorders that affect the structure or function of the brain or spinal cord, which together form the central nervous system (CNS).

[0085] In some embodiments, the CNS disorder is glioma. In some embodiments, the CNS disorder is glioblastoma. In some embodiments, the CNS disorder is epilepsy. In some embodiments, the CNS disorder is cortical dysplasia (e.g., focal cortical dysplasia). In some embodiments, the CNS disorder is selected from tuberous sclerosis, brain tumors, fragile X syndrome, Down syndrome, Rett syndrome, Alzheimer's disease, Parkinson's disease, and Huntington's disease.

[0086] In some embodiments, the CNS disorder is epilepsy. In some embodiments, the individual has undergone epilepsy surgery. In some embodiments, the individual has at least 5 seizures within 30 days after epilepsy surgery or has no seizure-free week after epilepsy surgery. In some embodiments, the method further comprises administering to the individual an effective amount of an anti-epileptic agent.

[0087] In some embodiments, the CNS disorder is glioblastoma. In some embodiments, the glioblastoma is recurrent glioblastoma. In some embodiments, the glioblastoma is newly diagnosed glioblastoma. In some embodiments, the individual has undergone surgical resection of the newly diagnosed glioblastoma before the start of nanoparticle administration.

[0088] Mitochondrial disorders

[0089] Mitochondria are organelles present in most eukaryotic cells. In addition to producing ATP, mitochondria are also involved in other cellular functions such as cell homeostasis, signaling pathways, and steroid synthesis.

[0090] Individuals with mitochondrial-related disorders (i.e., mitochondrial disorders), including but not limited to those with ataxia, kidney disorders, liver disorders, metabolic disorders, myopathies, neuropathies, spinal cord disorders, brain disorders, oxidative phosphorylation disorders, aging disorders, autism spectrum disorders, chronic inflammatory disorders, diabetes, and fatty acid oxidation disorders, can be treated with the methods described herein. In some embodiments, the individual with a mitochondrial-related disorder has a disorder associated with a mitochondrial DNA mutation. In some embodiments, the individual with a mitochondrial-related disorder has a disorder associated with an X chromosome mutation. In some embodiments, the individual with a mitochondrial-related disorder has a disorder associated with a nuclear DNA mutation. In some embodiments, the individual with a mitochondrial-related disorder has Leigh syndrome, such as maternally inherited Leigh syndrome. In some embodiments, Leigh syndrome is infantile Leigh syndrome, juvenile Leigh syndrome, or adult Leigh syndrome. In some embodiments, the individual with a mitochondrial-related disorder has MELAS syndrome. In some embodiments, the individual with a mitochondrial-related disorder has NARP syndrome.

[0091] Individuals with metabolic disorders, including but not limited to disorders related to cellular glucose consumption (e.g., abnormally high cellular glucose consumption in one or more tissues), disorders related to insulin resistance, hypoglycemia, hyperinsulinemic hypoglycemia, type 1 diabetes, type 2 diabetes, and metabolic syndrome, can be treated with the methods described herein.

[0092] The methods described herein can be used for any one or more of the following purposes: alleviating one or more symptoms of an individual with a mitochondrial-related disorder, reducing one or more symptoms of an individual with a mitochondrial-related disorder, preventing one or more symptoms of an individual with a mitochondrial-related disorder, treating one or more symptoms of an individual with a mitochondrial-related disorder, improving one or more symptoms of an individual with a mitochondrial-related disorder, and delaying the onset of one or more symptoms of an individual with a mitochondrial-related disorder.

[0093] As used herein, the terms "mitochondrial-related disorder" and "mitochondrial disorder" refer to any disease or disorder caused by mitochondrial dysfunction. Mitochondrial-related disorders can cause a variety of complex symptoms. Symptoms of mitochondrial-related disorders include, for example, muscle weakness, muscle cramps, seizures, food reflux, learning disabilities, deafness, short stature, ophthalmoplegia, diabetes, heart problems, and stroke-like episodes. The severity of the symptoms of mitochondrial-related disorders ranges from life-threatening to barely noticeable.

[0094] Individuals with mitochondrial-related disorders can be classified into one or more subsets of mitochondrial-related disorders based on genotype, phenotypic manifestations, and / or one or more symptoms. In some embodiments, an individual with a mitochondrial-related disorder has one or more of the following: ataxia, kidney disorder, liver disorder, metabolic disorder, myopathy, neuropathy, spinal cord disorder, brain disorder, oxidative phosphorylation disorder, aging disorder, autism spectrum disorder, chronic inflammatory disorder, or fatty acid oxidation disorder. In some embodiments, an individual with a mitochondrial-related disorder has one or more of the following: ataxia, kidney disorder, liver disorder, metabolic disorder, myopathy, neuropathy, spinal cord disorder, brain disorder, or oxidative phosphorylation disorder. In some embodiments, an individual with a mitochondrial-related disorder has one or more of the following: aging disorder, autism spectrum disorder, chronic inflammatory disorder, diabetes, or fatty acid oxidation disorder. In some embodiments, an individual with a mitochondrial-related disorder has at least ataxia. In some embodiments, an individual with a mitochondrial-related disorder has at least spinal cord disorder and brain disorder. In some embodiments, an individual with a mitochondrial-related disorder has at least neuropathy, spinal cord disorder, and brain disorder. In some embodiments, an individual with a mitochondrial-related disorder has at least myopathy and neuropathy.

[0095] Methods for treating cancer

[0096] The methods described herein can be used to treat an individual with cancer who has an mTOR activation aberration at one or more genes (such as TSC1, TSC2, RPS6, PTEN, TP53, RB1, ATRX, or FAT1). In some embodiments, there are methods for treating cancer in an individual with an mTOR activation aberration at TSC2. Based on having an mTOR activation aberration at one or more genes (e.g., TSC1, TSC2, RPS6, PTEN, TP53, RB1, ATRX, or FAT1), an individual with cancer can be selected for treatment by the methods described herein. In some embodiments, an individual is selected for treatment based on having an mTOR activation aberration at TSC2.

[0097] In some embodiments, cancer in an individual (e.g., advanced and / or malignant cancer, such as PEComa, e.g., advanced and / or malignant cancer, such as locally advanced inoperable cancer, e.g., solid tumor) can be treated by the methods described herein, including subcutaneously administering to the individual an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor and a carrier protein, wherein the individual is selected for treatment based on having an mTOR activation aberration at TSC2. In some embodiments, cancer in an individual (e.g., advanced and / or malignant cancer, such as PEComa, e.g., advanced and / or malignant cancer, such as locally advanced inoperable cancer, e.g., solid tumor) can be treated by the methods described herein, including subcutaneously administering to the individual an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor and a carrier protein, wherein the individual has an mTOR activation aberration at TSC2. In some embodiments, the mTOR activation aberration at TSC2 comprises a mutation in TSC2. In some embodiments, the mutation is selected from splice site mutation, nonsense mutation, frameshift mutation, and missense mutation. In some embodiments, the mTOR activation aberration at TSC2 comprises a single nucleotide variant (SNV). In some embodiments, the SNV comprises a mutation selected from C1503T, C2743G, C5383T, C3755G, G760T, C3442T, G880A, T707C, A4949G, or deletion of any one or more amino acids at positions 1405 - 1409, 1960 - 1970, 4999, 5002, 3521, 5208, 5238 - 5255. In some embodiments, the mTOR activation aberration at TSC2 comprises a copy number variation of TSC2. In some embodiments, the mTOR activation aberration at TSC2 is a loss-of-function mutation. In some embodiments, the mTOR activation aberration in TSC2 comprises an aberrant expression level of TSC2. In some embodiments, the mTOR activation aberration in TSC2 comprises an aberrant activity level of the protein encoded by TSC2. In some embodiments, the mTOR activation aberration in TSC2 comprises loss of heterozygosity of TSC2. In some embodiments, the mTOR inhibitor is a rapamycin drug. In some embodiments, the mTOR inhibitor is rapamycin or a derivative thereof. In some embodiments, the mTOR inhibitor is rapamycin. In some embodiments, the carrier protein is albumin (e.g., human serum albumin). In some embodiments, the dose of the mTOR inhibitor in the composition for each administration is about 0.1 mg / m 2 to about 100 mg / m 2 (e.g., about 0.1 mg / m 2 to about 10 mg / m 2 , about 10 mg / m 2 to about 50 mg / m 2 , about 50 mg / m2 to about 100 mg / m 2 、about 75 mg / m 2 to about 100 mg / m 2 ). In some embodiments, the method comprises subcutaneously administering the nanoparticle composition to the individual weekly for about two weeks, followed by a rest period of about one week. In some embodiments, the cancer is selected from pancreatic neuroendocrine cancer, endometrial cancer, breast cancer, lymphangioleiomyomatosis (LAM), prostate cancer, hepatocellular carcinoma, melanoma, renal cell carcinoma, bladder cancer, endometrial cancer, ovarian cancer, gynecological cancer, sarcoma, perivascular epithelioid cell neoplasm (PEComa), Hodgkin lymphoma, and multiple myeloma. In some embodiments, the cancer is PEComa. In some embodiments, regardless of the nature of the cancer, individuals are selected for treatment based on having a TSC2 aberration (e.g., a TSC2 mutation). In some embodiments, the individual does not have a TSC1 aberration (e.g., a TSC1 mutation).

[0098] In some embodiments, methods of treating cancer in an individual (e.g., advanced and / or malignant cancer, such as PEComa, e.g., advanced and / or malignant cancer, such as locally advanced inoperable cancer, e.g., solid tumor) are provided, including subcutaneously administering to the individual an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor and a carrier protein, wherein the individual is selected for treatment based on having a TSC2 aberration (e.g., a TSC2 mutation). In some embodiments, methods of treating cancer in an individual (e.g., advanced and / or malignant cancer, such as PEComa, e.g., advanced and / or malignant cancer, such as locally advanced inoperable cancer, e.g., solid tumor) are provided, including subcutaneously administering to the individual an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor and a carrier protein, wherein the individual is selected for treatment based on a) having a TSC2 aberration (e.g., a TSC2 mutation), and b) having an RPS6 aberration (e.g., an aberrant phosphorylation level of the protein encoded by RPS6 (e.g., phosphorylation at residues S235, S236, S240, and / or S244)). In some embodiments, methods of treating cancer in an individual (e.g., advanced and / or malignant cancer, such as PEComa, e.g., advanced and / or malignant cancer, such as locally advanced inoperable cancer, e.g., solid tumor) are provided, including subcutaneously administering to the individual an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor and a carrier protein, wherein the individual is selected for treatment based on a) having a TSC2 aberration (e.g., a TSC2 mutation), and b) not having a TSC1 mutation. In some embodiments, methods of treating cancer in an individual (e.g., advanced and / or malignant cancer, such as PEComa, e.g., advanced and / or malignant cancer, such as locally advanced inoperable cancer, e.g., solid tumor) are provided, including subcutaneously administering to the individual an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor and a carrier protein, wherein the individual is selected for treatment based on a) having a TSC2 aberration (e.g., a TSC2 mutation), b) not having a TSC1 mutation, and c) having an RPS6 aberration (e.g., an aberrant phosphorylation level of the protein encoded by RPS6) (e.g., phosphorylation at residues S235, S236, S240, and / or S244). In some embodiments, the mTOR activation aberration at RPS6 includes a positive state of phosphorylated S6 (pS6) (e.g., phosphorylation at residues S235, S236, S240, and / or S244). In some embodiments, the mutation is selected from splice site mutations, nonsense mutations, frameshift mutations, and missense mutations. In some embodiments, the mTOR inhibitor is a limus drug. In some embodiments, the mTOR inhibitor is rapamycin or a derivative thereof. In some embodiments, the mTOR inhibitor is rapamycin. In some embodiments, the carrier protein is albumin (e.g., human serum albumin).In some embodiments, the dose of the mTOR inhibitor in the composition per administration is about 0.1 mg / m². 2 to about 100 mg / m² 2 (e.g., about 0.1 mg / m² 2 to about 10 mg / m² 2 , about 10 mg / m² 2 to about 50 mg / m² 2 , about 50 mg / m² 2 to about 100 mg / m² 2 , about 75 mg / m² 2 to about 100 mg / m² 2 ). In some embodiments, the method includes subcutaneously administering the nanoparticle composition to the individual about two weeks per week, followed by a break of about one week. In some embodiments, the cancer is selected from pancreatic neuroendocrine cancer, endometrial cancer, breast cancer, lymphangioleiomyomatosis (LAM), prostate cancer, hepatocellular carcinoma, melanoma, renal cell carcinoma, bladder cancer, endometrial cancer, ovarian cancer, gynecological cancer, sarcoma, perivascular epithelioid cell neoplasm (PEComa), Hodgkin lymphoma, and multiple myeloma. In some embodiments, the cancer is PEComa. In some embodiments, regardless of the nature of the cancer, individuals are selected for treatment based on having TSC2 aberration and RPS6 aberration.

[0099] In some embodiments, there is provided a method of treating cancer (e.g., advanced and / or malignant cancer, such as PEComa, e.g., advanced and / or malignant cancer, such as locally advanced inoperable cancer, e.g., solid tumor) in an individual, comprising subcutaneously administering to the individual a composition comprising nanoparticles comprising rapamycin or a derivative thereof and albumin, wherein individuals are selected for treatment based on a) having a TSC2 aberration (e.g., a TSC2 mutation), and b) having an aberrant phosphorylation level of the protein encoded by RPS6 (e.g., phosphorylation at residues S235, S236, S240, and / or S244), wherein the dose of rapamycin or a derivative thereof in the composition per administration is about 0.1 mg / m² 2 to about 100 mg / m² 2 (e.g., about 0.1 mg / m² 2 to about 10 mg / m² 2 , about 10 mg / m² 2 to about 25 mg / m² 2 , about 25 mg / m² 2 to about 100 mg / m² 2 , about 50 mg / m² 2 to about 100 mg / m² 2 , about 75 mg / m² 2 to about 100 mg / m² 2), and wherein the composition is administered subcutaneously once a week for about two weeks, followed by a break of about one week.

[0100] In some embodiments, there is provided a method of treating cancer (e.g., advanced and / or malignant cancer, such as PEComa, e.g., advanced and / or malignant cancer, such as locally advanced inoperable cancer, e.g., solid tumors) in an individual, comprising administering subcutaneously to the individual a composition comprising nanoparticles comprising rapamycin or a derivative thereof and albumin, wherein the individual is selected for treatment based on a) having a TSC2 aberration (e.g., a TSC2 mutation), b) not having a TSC1 mutation, and c) having an aberrant phosphorylation level of the protein encoded by RPS6 (e.g., phosphorylation at residues S235, S236, S240, and / or S244), wherein the dose of rapamycin or a derivative thereof in the composition for each administration is about 10 mg / m 2 to about 100 mg / m 2 (e.g., about 25 mg / m 2 to about 100 mg / m 2 、about 50 mg / m 2 to about 100 mg / m 2 、about 75 mg / m 2 to about 100 mg / m 2 ), and wherein the composition is administered subcutaneously once a week for about two weeks, followed by a break of about one week.

[0101] In some embodiments, the aberrant phosphorylation level of the protein encoded by RPS6 is a positive state of phosphorylated S6 (pS6). In some embodiments, the aberrant phosphorylation level of the protein encoded by RPS6 is an increased phosphorylation of S6 in cancer compared to a reference tissue. In some embodiments, the reference tissue is derived from non-cancerous tissue of the individual. In some embodiments, the reference tissue is derived from the corresponding tissue of another individual not suffering from cancer.

[0102] In some embodiments, there is provided a method of treating a population of individuals suffering from different cancers (e.g., advanced and / or malignant cancers, such as locally advanced inoperable cancers, e.g., solid tumors), comprising administering subcutaneously to the population of individuals an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor (e.g., rapamycin) and a carrier protein (e.g., albumin), wherein each individual has a TSC2 aberration (e.g., a TSC2 mutation). In some embodiments, the individual does not have a TSC1 mutation.

[0103] In some embodiments, methods are provided for selecting an individual for treatment based on having cancer with a TSC2 mutation, wherein the treatment comprises subcutaneously administering to the individual a composition comprising nanoparticles comprising rapamycin or a derivative thereof and albumin, wherein optionally, the dose of rapamycin or a derivative thereof in the composition for each administration is from about 10 mg / m 2 to about 100 mg / m 2 (e.g., from about 25 mg / m 2 to about 100 mg / m 2 , from about 50 mg / m 2 to about 100 mg / m 2 , from about 75 mg / m 2 to about 100 mg / m 2 ), and wherein optionally the composition is administered subcutaneously once a week for about two weeks, followed by a rest period of about one week. In some embodiments, the individual does not have a TSC1 mutation.

[0104] The cancer treated by the methods contemplated in the present application can be any cancer having one or more mTOR activation aberrations at any gene selected from TSC1, TSC2, TP53, RB1, ATRX, FAT1, PTEN, and RPS6, and RPS6. In some embodiments, the cancer has one or more mTOR activation aberrations at any one gene selected from TSC1, TSC2, TP53, and RPS6. In some embodiments, the cancer has at least one mTOR activation aberration at RPS6 and at least one mTOR activation aberration at TSC1, TSC2, or TP53. In some embodiments, the cancer has at least one mTOR activation aberration at RPS6 and at least one mTOR activation aberration at TSC1 or TSC2.

[0105] In some embodiments, the cancer is a solid tumor. In some embodiments, the cancer is a hematological cancer.

[0106] In some embodiments, the cancer is advanced. In some embodiments, the cancer is malignant. In some embodiments, the cancer is locally advanced and inoperable.

[0107] In some embodiments, the cancer is selected from pancreatic neuroendocrine cancer, endometrial cancer, breast cancer, lymphangioleiomyomatosis (LAM), prostate cancer, hepatocellular carcinoma, melanoma, renal cell carcinoma, bladder cancer, endometrial cancer, ovarian cancer, gynecological cancer, sarcoma, perivascular epithelioid cell neoplasm (PEComa), Hodgkin lymphoma, and multiple myeloma.

[0108] In some embodiments, the cancer is PEComa. In some embodiments, the cancer is advanced PEComa. In some embodiments, the cancer is advanced and malignant PEComa. In some embodiments, the PEComa is primary uterine PEComa. In some embodiments, the PEComa is primary retroperitoneal PEComa. In some embodiments, the PEComa is primary renal PEComa. In some embodiments, the PEComa is primary pulmonary PEComa. In some embodiments, the PEComa is primary pelvic PEComa.

[0109] TSC2 is also known as tuberin, tuberous sclerosis 2 protein, protein phosphatase 1 regulatory subunit 160, TSC4, PPP1R160, and LAM. The TSC2 protein functions as part of a complex with TSC1 by negatively regulating mTORC1 signaling. In some embodiments, according to the GRCh38.p2 assembly of the human genome, the nucleic acid sequence of the wild-type TSC2 gene is identified by Genbank accession number NC_000016.10, from nucleotide 2047936 to nucleotide 2088712 on the plus strand of chromosome 16. The wild-type TSC2 gene contains 42 exons. Mutations in the TSC2 gene can occur in any one or any combination of the 42 exons, or in any intron or non-coding region of the TSC2 gene.

[0110] In some embodiments, the amino acid sequence of the wild-type TSC2 protein is identified by Genbank accession number NP_000539.2. In some embodiments, the amino acid sequence of the wild-type TSC2 protein is identified by Genbank accession number NP_001070651.1. In some embodiments, the amino acid sequence of the wild-type TSC2 protein is identified by Genbank accession number NP_001107854.1.

[0111] In some embodiments, the nucleic acid sequence of the cDNA encoding the wild-type TSC2 protein is identified by Genbank accession number NM_000548.3. In some embodiments, the nucleic acid sequence of the cDNA encoding the wild-type TSC2 protein is identified by Genbank accession number NM_001077183.1. In some embodiments, the nucleic acid sequence of the cDNA encoding the wild-type TSC2 protein is identified by Genbank accession number NM_001114382.1.

[0112] In some embodiments, individuals are selected for treatment based on having an mTOR activation aberration at TSC2. In some embodiments, the mTOR activation aberration at TSC2 includes a mutation in TSC2. In some embodiments, the mutation is selected from splice site mutations, nonsense mutations, frameshift mutations, and missense mutations. In some embodiments, the mTOR activation aberration at TSC2 includes a single nucleotide variant (SNV). In some embodiments, the SNV comprises a mutation selected from C1503T, C2743G, C5383T, C3755G, G760T, C3442T, G880A, T707C, A4949G, or a deletion of any one or more amino acids at positions 1405 - 1409, 1960 - 1970, 4999, 5002, 3521, 5208, 5238 - 5255.

[0113] In some embodiments, the mutation is a two - point mutation. In some embodiments, the mTOR activation aberration at TSC2 is a loss - of - function mutation. In some embodiments, the mTOR activation aberration at TSC2 includes a homozygous deletion. In some embodiments, the mTOR activation aberration of TSC2 includes a copy number variation of TSC2. In some embodiments, the mTOR activation aberration at TSC2 includes an aberrant expression level of TSC2. In some embodiments, the mTOR activation aberration at TSC2 includes an aberrant activity level of the protein encoded by TSC2.

[0114] Ribosomal protein S6 (RPS6) is also known as S6. Ribosomes are organelles that catalyze protein synthesis and consist of a small 40S subunit and a large 60S subunit. Together, these subunits are composed of 4 types of RNA and approximately 80 structurally different proteins. This gene encodes a cytoplasmic ribosomal protein that is a component of the 40S subunit. This protein belongs to the S6E family of ribosomal proteins. It is the major substrate of protein kinases in ribosomes and has a subset of five C - terminal serine residues that are phosphorylated by different protein kinases. Phosphorylation is induced by a variety of stimuli, including growth factors, tumor promoters, and mitogens. Dephosphorylation occurs during growth arrest. The protein can control cell growth and proliferation through the selective translation of specific classes of mRNA. This is typical for genes encoding ribosomal proteins, and multiple processed pseudogenes of this gene are dispersed throughout the genome.

[0115] In some embodiments, the nucleic acid sequence of the wild-type RPS6 gene is identified by Genbank accession number NC_000009.12, assembled according to GRCh38.p13 of the human genome, from nucleotide 19375715 to nucleotide 19380236 on the plus strand of chromosome 9. The wild-type RPS6 gene includes 6 exons. Mutations in the RPS6 gene can occur in any one or any combination of the 6 exons, or in any intron or non-coding region of the RPS6 gene.

[0116] In some embodiments, the amino acid sequence of the wild-type RPS6 protein is identified by Genbank accession number NM_001010.3.

[0117] In some embodiments, individuals are selected for treatment based on having an mTOR activation aberration at RPS6. In some embodiments, the mTOR activation aberration at RPS6 includes an aberrant phosphorylation level of the protein encoded by RPS6 (e.g., phosphorylation at residues S235, S236, S240, and / or S244). In some embodiments, the aberrant phosphorylation level of the protein encoded by RPS6 is a positive state of phosphorylated S6 (pS6). In some embodiments, the aberrant phosphorylation level of the protein encoded by RPS6 is an increased phosphorylation of S6 in cancer as compared to a reference tissue. In some embodiments, the reference tissue is derived from non-cancerous tissue of the individual. In some embodiments, the reference tissue is derived from the corresponding tissue of another individual who does not have cancer. The status of phosphorylated S6 can be evaluated by an antibody that binds to the phosphorylated residue in S6 by IHC staining (e.g., an antibody that detects the endogenous level of ribosomal protein S6 only when phosphorylated at Ser235 and 236). In some embodiments, the expression level of RPS6 is evaluated by immunohistochemistry. In some embodiments, the mTOR activation aberration at RPS6 includes an aberrant expression level of RPS6.

[0118] mTOR inhibitor

[0119] The methods described herein in some embodiments include subcutaneous administration of a nanoparticle composition of an mTOR inhibitor. mTOR is a serine / threonine-specific protein kinase downstream of the phosphatidylinositol 3-kinase (PI3K) / Akt (protein kinase B) pathway and a key regulator of cell survival, proliferation, stress, and metabolism. Dysregulation of the mTOR pathway has been found in many human cancers, and mTOR inhibition has a substantial inhibitory effect on tumor progression.

[0120] The mammalian target of rapamycin (mTOR) (also known as the mechanistic target of rapamycin or FK506-binding protein 12-rapamycin-associated protein 1 (FRAP1)) is an atypical serine / threonine protein kinase present in two distinct complexes, mTOR complex 1 (mTORC1) and mTOR complex 2 (mTORC2). mTORC1 consists of mTOR, regulatory-associated protein of mTOR (Raptor), mammalian lethal with SEC13 protein 8 (MLST8), PRAS40, and DEPTOR (Kim et al., (2002). Cell 110:163–75; Fang et al., (2001). Science 294(5548):1942–5). mTORC1 integrates four major signal inputs: nutrients (such as amino acids and phosphatidic acid), growth factors (insulin), energy, and stress (such as hypoxia and DNA damage). Amino acid availability signals to mTORC1 via a pathway involving Rag and Ragulator (LAMTOR1-3), and growth factors and hormones (e.g., insulin) signal to mTORC1 via Akt, which inactivates TSC2 to prevent inhibition of mTORC1. Alternatively, low ATP levels lead to AMPK-dependent activation of TSC2 and phosphorylation of raptor to reduce mTORC1 signaling proteins.

[0121] Active mTORC1 has many downstream biological effects, including translation of mRNA through phosphorylation of downstream targets (4E-BP1 and p70 S6 kinase), inhibition of autophagy (Atg13, ULK1), ribosome biogenesis, and transcriptional activation, leading to mitochondrial metabolism or adipogenesis. Thus, when conditions are favorable, mTORC1 activity promotes cell growth, or under stress or unfavorable conditions, mTORC1 activity promotes catabolic processes.

[0122] mTORC2 consists of mTOR, the rapamycin-insensitive mTOR partner (RICTOR), GβL, and mammalian stress-activated protein kinase interacting protein 1 (mSIN1). Compared with mTORC1, for which many upstream signals and cellular functions have been defined (see above), relatively little is known about mTORC2 biology. mTORC2 regulates cytoskeletal organization by stimulating F-actin stress fibers, paxillin, RhoA, Rac1, Cdc42, and protein kinase Cα (PKCα). It has been observed that knockdown of mTORC2 components affects actin polymerization and disrupts cell morphology (Jacinto et al., (2004). Nat. Cell Biol. 6, 1122-1128; Sarbassov et al., (2004). Curr. Biol. 14, 1296-1302). This suggests that mTORC2 controls the actin cytoskeleton by promoting protein kinase Cα (PKCα) phosphorylation, paxillin phosphorylation and its re-localization to focal adhesions, and GTP loading of RhoA and Rac1. The molecular mechanisms by which mTORC2 regulates these processes have not been determined.

[0123] In some embodiments, the mTOR inhibitor is an inhibitor of mTORC1. In some embodiments, the mTOR inhibitor is an inhibitor of mTORC2. In some embodiments, the mTOR inhibitor is an inhibitor of both mTORC1 and mTORC2.

[0124] In some embodiments, the mTOR inhibitor is a rapamycin drug. Examples of rapamycin drugs include, but are not limited to, rapamycin, temsirolimus (CCI-779), everolimus (RAD001), deforolimus (AP-23573), diflorasone (MK-8669), zotarolimus (ABT-578), pimecrolimus, and tacrolimus (FK-506). In some embodiments, the rapamycin drug is selected from temsirolimus (CCI-779), everolimus (RAD001), deforolimus (AP-23573), diflorasone (MK-8669), zotarolimus (ABT-578), pimecrolimus, and tacrolimus (FK-506). In some embodiments, the mTOR inhibitor is an mTOR kinase inhibitor, such as CC-115 or CC-223.

[0125] In some embodiments, the mTOR inhibitor is rapamycin. Rapamycin is a macrolide antibiotic that complexes with FKBP-12 and inhibits the mTOR pathway by binding to mTORC1.

[0126] In some embodiments, the mTOR inhibitor is selected from rapamycin (sirolimus), BEZ235 (NVP-BEZ235), everolimus (also known as RAD001, Zortress, Certican and Afinitor), AZD8055, temsirolimus (also known as CCI-779 and Torisel), CC-115, CC-223, PI-103, Ku-0063794, INK 128, AZD2014, NVP-BGT226, PF-04691502, CH5132799, GDC-0980 (RG7422), Torin 1, WAY-600, WYE-125132, WYE-687, GSK2126458, PF-05212384 (PKI-587), PP-121, OSI-027, Palomid529, PP242, XL765, GSK1059615, WYE-354, and deforolimus (also known as deforomomycin).

[0127] BEZ235 (NVP - BEZ235) is an imidazo - quinoline derivative and is a catalytic inhibitor of mTORC1 (Roper J et al., PLoS One, 2011, 6(9), e25132). Everolimus is a 40 - O-(2 - hydroxyethyl) derivative of rapamycin and binds to the cyclophilin FKBP - 12, and this complex is also mTORC1. AZD8055 is a small molecule that inhibits the phosphorylation of mTORC1 (p70S6K and 4E - BP1). Temsirolimus is a small molecule that forms a complex with the FK506 - binding protein and blocks its activation when mTOR resides in the mTORC1 complex. PI - 103 is a small molecule that inhibits the activation of the rapamycin - sensitive (mTORC1) complex (Knight et al., (2006) Cell. 125:733 - 47). KU - 0063794 is a small molecule that inhibits the phosphorylation of mTORC1 at Ser2448 in a dose - dependent and time - dependent manner. INK 128, AZD2014, NVP - BGT226, CH5132799, WYE - 687 are small molecule inhibitors of mTORC1 respectively. PF - 04691502 inhibits mTORC1 activity. GDC - 0980 is an orally bioavailable small molecule that inhibits class I PI3 kinases and TORC1. Torin 1 is a potent small molecule inhibitor of mTOR. WAY - 600 is a potent, ATP - competitive and selective inhibitor of mTOR. WYE - 125132 is an ATP - competitive small molecule inhibitor of mTORC1. GSK2126458 is an inhibitor of mTORC1. PKI - 587 is a potent dual inhibitor of PI3Kα, PI3Kγ and mTOR. PP - 121 is a multi - target inhibitor of PDGFR, Hck, mTOR, VEGFR2, Src and Abl. OSI - 027 is a selective and potent dual inhibitor of mTORC1 and mTORC2 with IC50 values of 22 nM and 65 nM respectively. Palomid 529 is a small molecule inhibitor of mTORC1 that lacks affinity for ABCB1 / ABCG2 and has good brain permeability (Lin et al., (2013) Int J Cancer DOI:10.1002 / ijc.28126 (e - published before print)). PP242 is a selective mTOR inhibitor. XL765 is a dual inhibitor of mTOR / PI3k against mTOR, p110α, p110β, p110γ and p110δ. GSK1059615 is a novel and dual inhibitor of PI3Kα, PI3Kβ, PI3Kδ, PI3Kγ and mTOR. WYE - 354 inhibits mTORC1 in HEK293 cells (0.2 μM–5 μM) and HUVEC cells (10 nM - 1 μM).WYE-354 is a potent, specific, and ATP-competitive inhibitor of mTOR. Deforolimus (AP23573, MK-8669) is a selective mTOR inhibitor.

[0128] Nanoparticle composition

[0129] The mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising (and in various embodiments consisting essentially of or consisting of) an mTOR inhibitor (such as rapamycin) and albumin (such as human serum albumin). Nanoparticles of poorly water-soluble drugs (such as macrolides) have been disclosed in, for example, US5916596 A; US6506405B1; US 6749868 B1, US 6537579 B1, US 7820788 B2, and US 8911786 B2, as well as US2006 / 0263434A1, US2007 / 0082838A1, and WO2008 / 137148A2, each of which is incorporated herein by reference in its entirety.

[0130] The present disclosure describes a composition, such as a pharmaceutical composition, that includes nanoparticles comprising an mTOR inhibitor and albumin. The mTOR inhibitor is an agent selected from compounds that inhibit mammalian target of rapamycin (mTOR). In some embodiments, the mTOR inhibitor is rapamycin (also known as sirolimus) or an analogue thereof. In some embodiments, the mTOR inhibitor is a ridaforolimus drug, which includes rapamycin and its analogues. Examples of ridaforolimus drugs include, but are not limited to, temsirolimus (CCI-779), everolimus (RAD001), deforolimus (AP-23573), deforolimus (MK-8669), zotarolimus (ABT-578), pimecrolimus, and tacrolimus (FK-506). In some embodiments, the ridaforolimus drug is selected from temsirolimus (CCI-779), everolimus (RAD001), deforolimus (AP-23573), deforolimus (MK-8669), zotarolimus (ABT-578), pimecrolimus, and tacrolimus (FK-506). In some embodiments, the mTOR inhibitor is an mTOR kinase inhibitor, such as CC-115 or CC-223. In some embodiments, the mTOR inhibitor is selected from rapamycin (sirolimus), BEZ235 (NVP-BEZ235), everolimus (also known as RAD001, Zortress, Certican, and Afinitor), AZD8055, temsirolimus (also known as CCI-779 and Torisel), CC-115, CC-223, PI-103, Ku-0063794, INK 128, AZD2014, NVP-BGT226, PF-04691502, CH5132799, GDC-0980 (RG7422), Torin 1, WAY-600, WYE-125132, WYE-687, GSK2126458, PF-05212384 (PKI-587), PP-121, OSI-027, Palomid 529, PP242, XL765, GSK1059615, WYE-354, and deforolimus (also known as deforolimus).

[0131] In some embodiments, the pharmaceutical composition further comprises one or more reagents for enhancing the dissolution of the composition in dry form and / or enhancing the stability of the composition. In some embodiments, the additional one or more reagents comprise sugars. The sugars can be, but are not limited to, monosaccharides, disaccharides, polysaccharides, and derivatives or modifications thereof. The sugar can be, for example, any one of mannitol, sucrose, fructose, lactose, maltose, glucose, or trehalose. In some embodiments, the additional one or more reagents comprise glycine. Thus, in one aspect, the present application provides a pharmaceutical composition suitable for subcutaneous administration to an individual, the pharmaceutical composition comprising a) nanoparticles comprising an mTOR inhibitor (such as rapamycin) and albumin, and b) a sugar.

[0132] In some embodiments, the sugar is present in an amount that effectively increases the stability of the nanoparticles in the composition compared to a nanoparticle composition without sugar. In some embodiments, the amount of sugar can effectively improve the filterability of the nanoparticle composition compared to a composition without sugar.

[0133] In some embodiments, the sugar is present in an amount that effectively increases the solubility of the pharmaceutical composition. In some embodiments, the increased solubility includes an increased dissolution rate of the nanoparticle composition in dry form after addition of the reconstitution solution.

[0134] In some embodiments, when the nanoparticle composition is administered subcutaneously, the sugar is present in an amount that reduces the incidence or severity of side effects after administration. For example, in some embodiments, the side effect is a rash, and the composition comprises nanoparticles comprising an mTOR inhibitor and albumin, and the sugar is present in an amount that reduces the incidence of the rash after subcutaneous administration of the nanoparticle composition.

[0135] In some embodiments, the pharmaceutical composition comprises nanoparticles comprising an mTOR inhibitor and albumin, wherein the weight ratio of albumin to the mTOR inhibitor in the composition is from about 0.01:1 to about 100:1. In some embodiments, the composition comprises nanoparticles comprising an mTOR inhibitor (such as rapamycin) and albumin, wherein the weight ratio of albumin to the mTOR inhibitor (such as rapamycin) in the composition is about 18:1 or lower (including, for example, any one of about 1:1 to about 18:1, about 2:1 to about 15:1, about 3:1 to about 12:1, about 4:1 to about 10:1, about 5:1 to about 9:1, and about 9:1). In some embodiments, the composition comprises nanoparticles comprising rapamycin or a derivative thereof and albumin, wherein the weight ratio of albumin to rapamycin or a derivative thereof in the composition is about 18:1 or lower (including, for example, any one of about 1:1 to about 18:1, about 2:1 to about 15:1, about 3:1 to about 12:1, about 4:1 to about 10:1, about 5:1 to about 9:1, and about 9:1). In some embodiments, the mTOR inhibitor (such as rapamycin) is coated with albumin.

[0136] In some embodiments, the particles (such as nanoparticles) described herein have an average diameter or mean diameter that is no greater than any one of about 1000, 900, 800, 700, 600, 500, 400, 300, 200, 150, 120, and 100 nm. In some embodiments, the average diameter or mean diameter of the particles is no greater than about 200 nm. In some embodiments, the average diameter or mean diameter of the particles is between about 20 nm and about 400 nm. In some embodiments, the average diameter or mean diameter of the particles is between about 40 nm and about 200 nm. In some embodiments, the average diameter or mean diameter of the nanoparticles is about 100 - 120 nm, such as about 100 nm. In some embodiments, the average mean diameter of the particles is less than or equal to 120 nm. In some embodiments, the average mean diameter of the particles is about 100 - 120 nm, such as about 100 nm. In some embodiments, the particles are sterile-filterable. Methods for determining the average particle size are known in the art. For example, dynamic light scattering (DLS) has been routinely used to determine particle sizes based on the sub-micron scale. International Standard ISO22412 Particle Size Analysis - Dynamic Light Scattering, International Organization for Standardization (ISO) 2008 and the defined general terms for dynamic light scattering, Malvern Instruments Limited, 2011. In some embodiments, the particle size is measured as the volume-weighted mean particle size (Dv50) of the nanoparticles in the composition.

[0137] The compositions described herein can be stable aqueous suspensions of mTOR inhibitors, such as stable aqueous suspensions having an mTOR inhibitor concentration of any one of about 0.1 to about 200 mg / ml, about 0.1 to about 150 mg / ml, about 0.1 to about 100 mg / ml, about 0.1 to about 50 mg / ml, about 0.1 to about 20 mg / ml, about 1 to about 10 mg / ml, about 2 mg / ml to about 8 mg / ml, about 4 to about 6 mg / ml, and about 5 mg / ml. In some embodiments, the concentration of the mTOR inhibitor is at least any one of about 0.2 mg / ml, 1.3 mg / ml, 1.5 mg / ml, 2 mg / ml, 3 mg / ml, 4 mg / ml, 5 mg / ml, 6 mg / ml, 7 mg / ml, 8 mg / ml, 9 mg / ml, 10 mg / ml, 15 mg / ml, 20 mg / ml, 25 mg / ml, 30 mg / ml, 40 mg / ml, 50 mg / ml, 100 mg / ml, 150 mg / ml, or 200 mg / ml.

[0138] In some embodiments, the composition is a dry (e.g., lyophilized) composition that can be reconstituted, resuspended, or rehydrated to form a stable aqueous suspension of nanoparticles that typically comprise an mTOR inhibitor and albumin. In some embodiments, the composition is a liquid (e.g., aqueous) composition obtained by reconstituting or resuspending the dry composition. In some embodiments, the composition is an intermediate liquid (e.g., aqueous) composition that can be dried (e.g., lyophilized).

[0139] In some embodiments, nanoparticles comprising an mTOR inhibitor (such as rapamycin) are associated (e.g., coated) with albumin (such as human albumin or human serum albumin). In some embodiments, the composition comprises nanoparticles and mTOR inhibitor (such as rapamycin) in non-nanoparticle form (e.g., in solution form or in the form of a soluble albumin / nanoparticle complex), wherein at least any one of about 50%, 60%, 70%, 80%, 90%, 95%, or 99% of the mTOR inhibitor in the composition is in nanoparticle form. In some embodiments, the mTOR inhibitor (such as rapamycin) in the nanoparticles constitutes more than any one of about 50%, 60%, 70%, 80%, 90%, 95%, or 99% by weight of the nanoparticles. In some embodiments, the nanoparticles have a non-polymeric matrix. In some embodiments, the nanoparticles comprise a core of mTOR inhibitor (such as rapamycin) that is substantially free of polymeric material (such as a polymeric matrix).

[0140] In some embodiments, the composition includes albumin in both the nanoparticle and non-nanoparticle portions of the composition, wherein at least about 50%, 60%, 70%, 80%, 90%, 95%, or 99% of the albumin in the composition is in the non-nanoparticle portion of the composition.

[0141] In some embodiments, the weight ratio of albumin (such as human albumin or human serum albumin) to mTOR inhibitor (such as rapamycin) in the mTOR inhibitor nanoparticle composition is about 18:1 or less, such as about 15:1 or less, for example about 10:1 or less. In some embodiments, the weight ratio of albumin (such as human albumin or human serum albumin) to mTOR inhibitor (such as rapamycin) in the composition falls within the range of any one of about 1:1 to about 18:1, about 2:1 to about 15:1, about 3:1 to about 13:1, about 4:1 to about 12:1, about 5:1 to about 10:1. In some embodiments, the weight ratio of albumin to mTOR inhibitor (such as rapamycin) in the nanoparticle portion of the composition is any one of about 1:2, 1:3, 1:4, 1:5, 1:9, 1:10, 1:15, or less. In some embodiments, the weight ratio of albumin (such as human albumin or human serum albumin) to mTOR inhibitor (such as rapamycin) in the composition is any of the following: about 1:1 to about 18:1, about 1:1 to about 15:1, about 1:1 to about 12:1, about 1:1 to about 10:1, about 1:1 to about 9:1, about 1:1 to about 8:1, about 1:1 to about 7:1, about 1:1 to about 6:1, about 1:1 to about 5:1, about 1:1 to about 4:1, about 1:1 to about 3:1, about 1:1 to about 2:1, about 1:1 to about 1:1.

[0142] The nanoparticles described herein can be in a dry formulation (such as a lyophilized composition) or suspended in a biocompatible medium, such as a reconstitution solution. Suitable biocompatible media include, but are not limited to, water, buffered aqueous media, saline, buffered saline, optionally buffered solutions of amino acids, optionally buffered solutions of proteins, optionally buffered solutions of sugars, optionally buffered solutions of vitamins, optionally buffered solutions of synthetic polymers, lipid-containing emulsions, and the like.

[0143] In some embodiments, the pharmaceutically acceptable carrier includes albumin (such as human albumin or human serum albumin). Albumin can be of natural origin or synthetically prepared. In some embodiments, albumin is human albumin or human serum albumin. In some embodiments, albumin is recombinant albumin.

[0144] Human serum albumin (HSA) is M rA 65K highly soluble globular protein, consisting of 585 amino acids. HSA is the most abundant protein in plasma and accounts for 70 - 80% of the human plasma colloid osmotic pressure. The amino acid sequence of HSA contains a total of 17 disulfide bonds, a free thiol (Cys 34) and a single tryptophan (Trp 214). Intravenous administration of HSA solutions has been shown to prevent and treat hypovolemic shock (see, e.g., Tullis, JAMA, 237:355 - 360, 460 - 463, (1977) and Houser et al., Surgery, Gynecology and Obstetrics, 150:811 - 816 (1980)) and to be used in combination with exchange transfusion for the treatment of neonatal hyperbilirubinemia (see, e.g., Finlayson, Seminars in Thrombosis and Hemostasis, 6, 85 - 120, (1980)). Other albumins, such as bovine serum albumin, have been considered. The use of such non - human albumins may be appropriate, for example, in cases where these compositions are to be used in non - human mammals, such as in veterinary (including domestic pets and agricultural settings). Human serum albumin (HSA) has multiple hydrophobic binding sites (a total of 8 for fatty acids, which are endogenous ligands of HSA) and binds multiple groups of drugs, especially neutral and negatively charged hydrophobic compounds (Goodman et al., The Pharmacological Basis of Therapeutics, 9th ed., McGraw - Hill New York (1996)). Two high - affinity binding sites have been proposed in subdomains IIA and IIIA of HSA, which are highly elongated hydrophobic pockets with charged lysine and arginine residues near the surface, which act as attachment points for polar ligand features (see, e.g., Fehske et al Biochem.Pharmcol., 30, 687 - 92 (198a), Vorum, Dan.Med.Bull., 46, 379 - 99 (1999), Kragh - Hansen, Dan.Med.Bull., 1441, 131 - 40 (1990), Curry et al., Nat.Struct.Biol., 5, 827 - 35 (1998), Sugio et al., Protein.Eng., 12, 439 - 46 (1999), He et al., Nature, 358, 209 - 15 (199b) and Carter et al., Adv.Protein.Chem., 45, 153 - 203 (1994)).Rapamycin and propofol have been shown to bind to HSA (see, e.g., Paal et al., Eur. J. Biochem., 268(7), 2187 - 91(200a), Purcell et al., Biochem. Biophys. Acta, 1478(a), 61 - 8(2000), Altmayer et al., Arzneimittelforschung, 45, 1053 - 6(1995) and Garrido et al., Rev. Esp. Anestesiol. Reanim., 41, 308 - 12(1994)).

[0145] In some embodiments, the compositions described herein are substantially free (such as free) of surfactants, such as hydrogenated castor oil (or polyoxyethylated castor oil, including Cremophor (BASF)). In some embodiments, the mTOR inhibitor nanoparticle compositions (such as rapamycin / albumin nanoparticle compositions) are substantially free (such as free) of surfactants. When an mTOR inhibitor nanoparticle composition (such as a rapamycin / albumin nanoparticle composition) is administered to an individual, the composition is "substantially free of Cremophor" or "substantially free of surfactants" if the amount of Cremophor or surfactant in the composition is not sufficient to cause one or more side effects in the individual. In some embodiments, the mTOR inhibitor nanoparticle compositions (such as rapamycin / albumin nanoparticle compositions) contain less than about 20%, 15%, 10%, 7.5%, 5%, 2.5% or 1% of any of organic solvents or surfactants. In some embodiments, the albumin is human albumin or human serum albumin. In some embodiments, the albumin is recombinant albumin.

[0146] The amount of albumin in the compositions described herein will vary depending on the other components in the composition. In some embodiments, the composition includes an amount of albumin sufficient to stabilize the mTOR inhibitor (such as rapamycin) in an aqueous suspension, e.g., in the form of a stable colloidal suspension (such as a stable nanoparticle suspension). In some embodiments, the albumin is an amount that reduces the deposition rate of the mTOR inhibitor (such as rapamycin) in an aqueous medium. For particulate-containing compositions, the amount of albumin also depends on the size and density of the nanoparticles of the mTOR inhibitor.

[0147] An mTOR inhibitor (such as rapamycin) is "stable" in an aqueous suspension if it remains suspended in an aqueous medium (such as without visible precipitate or sediment) for an extended period of time, such as at least about any one of 0.1, 0.2, 0.25, 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 24, 36, 48, 60, or 72 hours. The suspension is generally but not necessarily suitable for administration to an individual (such as a human). The stability of the suspension is generally (but not necessarily) evaluated at a storage temperature (such as room temperature (such as 20 - 25 °C) or refrigerated conditions (such as 4 °C)). For example, if the suspension does not exhibit visible flocculation or particle aggregation to the naked eye or when observed using a 1000-fold optical microscope within about fifteen minutes after preparation, it is stable at the storage temperature. Stability can also be evaluated under accelerated test conditions, such as at a temperature of about 40 °C or higher.

[0148] In some embodiments, albumin is present in an amount sufficient to stabilize an mTOR inhibitor (such as rapamycin) at a certain concentration in an aqueous suspension. For example, the concentration of the mTOR inhibitor (such as rapamycin) in the composition is from about 0.1 to about 100 mg / ml, including for example any one of about 0.1 to about 50 mg / ml, about 0.1 to about 20 mg / ml, about 1 to about 10 mg / ml, about 2 mg / ml to about 8 mg / ml, about 4 to about 6 mg / ml, or about 5 mg / ml. In some embodiments, the concentration of the mTOR inhibitor (such as rapamycin) is at least about any one of 1.3 mg / ml, 1.5 mg / ml, 2 mg / ml, 3 mg / ml, 4 mg / ml, 5 mg / ml, 6 mg / ml, 7 mg / ml, 8 mg / ml, 9 mg / ml, 10 mg / ml, 15 mg / ml, 20 mg / ml, 25 mg / ml, 30 mg / ml, 40 mg / ml, and 50 mg / ml. In some embodiments, albumin is present in an amount to avoid the use of a surfactant (such as Cremophor) so that the composition is free or substantially free of a surfactant (such as Cremophor).

[0149] In some embodiments, the liquid form of the composition comprises from about 0.1% to about 50% (w / v) (e.g., about 0.5% (w / v), about 5% (w / v), about 10% (w / v), about 15% (w / v), about 20% (w / v), about 30% (w / v), about 40% (w / v), or about 50% (w / v)) of albumin. In some embodiments, the liquid form of the composition comprises from about 0.5% to about 5% (w / v) of albumin.

[0150] In some embodiments, the weight ratio of albumin to the mTOR inhibitor (such as rapamycin) in the mTOR inhibitor nanoparticle composition is such that a sufficient amount of the mTOR inhibitor binds to cells or is transported by cells. Although the weight ratio of albumin to the mTOR inhibitor (such as rapamycin) must be optimized for different combinations of albumin and mTOR inhibitor, generally the weight ratio (w / w) of albumin to the mTOR inhibitor (such as rapamycin) is from about 0.01:1 to about 100:1, about 0.02:1 to about 50:1, about 0.05:1 to about 20:1, about 0.1:1 to about 20:1, about 1:1 to about 18:1, about 2:1 to about 15:1, about 3:1 to about 12:1, about 4:1 to about 10:1, about 5:1 to about 9:1 or about 9:1. In some embodiments, the weight ratio of albumin to the mTOR inhibitor (such as rapamycin) is about 18:1 or less, 15:1 or less, 14:1 or less, 13:1 or less, 12:1 or less, 11:1 or less, 10:1 or less, 9:1 or less, 8:1 or less, 7:1 or less, 6:1 or less, 5:1 or less, 4:1 or less and 3:1 or less. In some embodiments, the weight ratio of albumin (such as human albumin or human serum albumin) to the mTOR inhibitor (such as rapamycin) in the composition is any of the following: about 1:1 to about 18:1, about 1:1 to about 15:1, about 1:1 to about 12:1, about 1:1 to about 10:1, about 1:1 to about 9:1, about 1:1 to about 8:1, about 1:1 to about 7:1, about 1:1 to about 6:1, about 1:1 to about 5:1, about 1:1 to about 4:1, about 1:1 to about 3:1, about 1:1 to about 2:1, about 1:1 to about 1:1.

[0151] In some embodiments, albumin allows the composition to be administered to an individual (such as a human) without significant side effects. In some embodiments, the amount of albumin (such as human serum albumin or human albumin) can effectively reduce one or more side effects of subcutaneous administration of the mTOR inhibitor (such as rapamycin) to a human. The term "reduce one or more side effects of administration, such as subcutaneous administration of the mTOR inhibitor (such as rapamycin)" refers to reducing, alleviating, eliminating or avoiding one or more undesirable effects caused by the mTOR inhibitor, as well as side effects caused by the delivery vehicle used to deliver the mTOR inhibitor (such as a solvent that makes the rapamycin drug suitable for injection). Such side effects include, for example, myelosuppression, neurotoxicity, hypersensitivity, inflammation, venous irritation, phlebitis, pain, skin irritation, peripheral neuropathy, febrile neutropenia, allergic reaction, venous thrombosis, extravasation and combinations thereof. However, these side effects are merely exemplary, and other side effects or combinations of side effects associated with the rapamycin drug (such as rapamycin) can be reduced.

[0152] In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising an mTOR inhibitor (such as rapamycin) and albumin (such as human albumin or human serum albumin), wherein the average diameter of the nanoparticles is no greater than about 200 nm. In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising an mTOR inhibitor (such as rapamycin) and albumin (such as human albumin or human serum albumin), wherein the average diameter of the nanoparticles is no greater than about 150 nm. In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising an mTOR inhibitor (such as rapamycin) and albumin (such as human albumin or human serum albumin), wherein the average diameter of the nanoparticles is no greater than about 150 nm (e.g., about 100 nm). In some embodiments, the average or mean diameter of the nanoparticles is about 100 - 120 nm, such as about 100 nm. In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising rapamycin and human albumin (such as human serum albumin), wherein the average diameter of the nanoparticles is no greater than about 150 nm (e.g., about 100 nm). In some embodiments, the average or mean diameter of the nanoparticles is about 100 - 120 nm, such as about 100 nm. In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising rapamycin and human albumin (such as human serum albumin), wherein the average or mean diameter of the nanoparticles is from about 10 to about 150 nm. In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising rapamycin and human albumin (such as human serum albumin), wherein the average or mean diameter of the nanoparticles is from about 40 to about 120 nm.

[0153] In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising an mTOR inhibitor (such as rapamycin) and albumin (such as human albumin or human serum albumin), wherein the composition further includes a sugar, and wherein the average diameter of the nanoparticles is no greater than about 200 nm. In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising an mTOR inhibitor (such as rapamycin) and albumin (such as human albumin or human serum albumin), wherein the composition further includes a sugar, and wherein the average diameter of the nanoparticles is no greater than about 150 nm. In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising an mTOR inhibitor (such as rapamycin) and albumin (such as human albumin or human serum albumin), wherein the composition further includes a sugar, and wherein the average diameter of the nanoparticles is no greater than about 150 nm (e.g., about 100 nm). In some embodiments, the average or mean diameter of the nanoparticles is about 100 - 120 nm, e.g., about 100 nm. In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising rapamycin and human albumin (such as human serum albumin), wherein the composition further includes a sugar, and wherein the average diameter of the nanoparticles is no greater than about 150 nm (e.g., about 100 nm). In some embodiments, the average or mean diameter of the nanoparticles is about 100 - 120 nm, e.g., about 100 nm. In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising rapamycin and human albumin (such as human serum albumin), wherein the composition further includes a sugar, and wherein the average or mean diameter of the nanoparticles is from about 10 to about 150 nm. In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising rapamycin and human albumin (such as human serum albumin), wherein the average or mean diameter of the nanoparticles is from about 40 to about 120 nm. In some embodiments, the average or mean diameter of the nanoparticles is about 100 - 120 nm, e.g., about 100 nm.

[0154] In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising an mTOR inhibitor (such as rapamycin) and albumin (such as human albumin or human serum albumin), wherein the average diameter of the nanoparticles is not greater than about 200 nm, and wherein the weight ratio of albumin to mTOR inhibitor in the composition is not greater than about 9:1 (such as about 9:1 or about 8:1). In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising an mTOR inhibitor (such as rapamycin) and albumin (such as human albumin or human serum albumin), wherein the average diameter of the nanoparticles is not greater than about 150 nm, and wherein the weight ratio of albumin to mTOR inhibitor in the composition is not greater than about 9:1 (such as about 9:1 or about 8:1). In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising rapamycin and human albumin (such as human serum albumin), wherein the average diameter of the nanoparticles is not greater than about 150 nm (e.g., about 100 nm), and wherein the weight ratio of albumin to mTOR inhibitor in the composition is about 9:1 or about 8:1. In some embodiments, the average or mean diameter of the nanoparticles is from about 10 nm to about 150 nm. In some embodiments, the average or mean diameter of the nanoparticles is from about 40 nm to about 120 nm. In some embodiments, the average or mean diameter of the nanoparticles is about 100 - 120 nm, such as about 100 nm.

[0155] In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising an mTOR inhibitor (such as rapamycin) and albumin (such as human albumin or human serum albumin), wherein the composition further includes a sugar, wherein the average diameter of the nanoparticles is not greater than about 200 nm, and wherein the weight ratio of albumin to mTOR inhibitor in the composition is not greater than about 9:1 (such as about 9:1 or about 8:1). In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising an mTOR inhibitor (such as rapamycin) and albumin (such as human albumin or human serum albumin), wherein the composition further includes a sugar, wherein the average diameter of the nanoparticles is not greater than about 150 nm, and wherein the weight ratio of albumin to mTOR inhibitor in the composition is not greater than about 9:1 (such as about 9:1 or about 8:1). In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising an mTOR inhibitor (such as rapamycin) and albumin (such as human albumin or human serum albumin), wherein the composition further includes a sugar, wherein the average diameter of the nanoparticles is about 150 nm, and wherein the weight ratio of albumin to mTOR inhibitor in the composition is not greater than about 9:1 (such as about 9:1 or about 8:1). In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising rapamycin and human albumin (such as human serum albumin), wherein the composition further includes a sugar, wherein the average diameter of the nanoparticles is not greater than about 150 nm (e.g., about 100 nm), and wherein the weight ratio of albumin to mTOR inhibitor in the composition is about 9:1 or about 8:1. In some embodiments, the average or mean diameter of the nanoparticles is from about 10 nm to about 150 nm. In some embodiments, the average or mean diameter of the nanoparticles is from about 40 nm to about 120 nm. In some embodiments, the average or mean diameter of the nanoparticles is about 100 - 120 nm, such as about 100 nm.

[0156] In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising an mTOR inhibitor (such as rapamycin) associated (e.g., coated) with albumin (such as human albumin or human serum albumin). In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising an mTOR inhibitor (such as rapamycin) associated (e.g., coated) with albumin (such as human albumin or human serum albumin), wherein the average diameter of the nanoparticles is no greater than about 200 nm. In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising an mTOR inhibitor (such as rapamycin) associated (e.g., coated) with albumin (such as human albumin or human serum albumin), wherein the average diameter of the nanoparticles is no greater than about 150 nm. In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising an mTOR inhibitor (such as rapamycin) associated (e.g., coated) with albumin (such as human albumin or human serum albumin), wherein the average diameter of the nanoparticles is from about 10 nm to about 150 nm. In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising an mTOR inhibitor (such as rapamycin) associated (e.g., coated) with human albumin (such as human serum albumin), wherein the average diameter of the nanoparticles is no greater than about 150 nm (e.g., about 100 nm). In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising an mTOR inhibitor (such as rapamycin) associated (e.g., coated) with human albumin (such as human serum albumin), wherein the average diameter of the nanoparticles is from about 10 nm to about 150 nm. In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising an mTOR inhibitor (such as rapamycin) associated (e.g., coated) with human albumin (such as human serum albumin), wherein the average diameter of the nanoparticles is from about 40 nm to about 120 nm. In some embodiments, the average or mean diameter of the nanoparticles is about 100 - 120 nm, such as about 100 nm.

[0157] In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising an mTOR inhibitor (such as rapamycin) associated (e.g., coated) with albumin (such as human albumin or human serum albumin), wherein the composition further includes a sugar. In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising an mTOR inhibitor (such as rapamycin) associated (e.g., coated) with albumin (such as human albumin or human serum albumin), wherein the composition further includes a sugar, wherein the average diameter of the nanoparticles is not greater than about 200 nm. In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising an mTOR inhibitor (such as rapamycin) associated (e.g., coated) with albumin (such as human albumin or human serum albumin), wherein the composition further includes a sugar, wherein the average diameter of the nanoparticles is not greater than about 150 nm. In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising an mTOR inhibitor (such as rapamycin) associated (e.g., coated) with albumin (such as human albumin or human serum albumin), wherein the composition further includes a sugar, wherein the average diameter of the nanoparticles is from about 10 nm to about 150 nm. In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising an mTOR inhibitor (such as rapamycin) associated (e.g., coated) with human albumin (such as human serum albumin), wherein the composition further includes a sugar, wherein the average diameter of the nanoparticles is not greater than about 150 nm (e.g., about 100 nm). In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising an mTOR inhibitor (such as rapamycin) associated (e.g., coated) with human albumin (such as human serum albumin), wherein the composition further includes a sugar, wherein the average diameter of the nanoparticles is from about 10 nm to about 150 nm. In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising an mTOR inhibitor (such as rapamycin) associated (e.g., coated) with human albumin (such as human serum albumin), wherein the composition further includes a sugar, wherein the average diameter of the nanoparticles is from about 40 nm to about 120 nm. In some embodiments, the average or mean diameter of the nanoparticles is about 100 - 120 nm, such as about 100 nm.

[0158] In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising an mTOR inhibitor (such as rapamycin) associated (e.g., coated) with albumin (such as human albumin or human serum albumin), wherein the weight ratio of albumin to mTOR inhibitor in the composition is not greater than about 9:1 (such as about 9:1 or about 8:1). In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising an mTOR inhibitor (such as rapamycin) associated (e.g., coated) with albumin (such as human albumin or human serum albumin), wherein the average diameter of the nanoparticles is not greater than about 200 nm, and wherein the weight ratio of albumin to mTOR inhibitor in the composition is not greater than about 9:1 (such as about 9:1 or about 8:1). In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising an mTOR inhibitor (such as rapamycin) associated (e.g., coated) with albumin (such as human albumin or human serum albumin), wherein the average diameter of the nanoparticles is not greater than about 150 nm, and wherein the weight ratio of albumin to mTOR inhibitor in the composition is not greater than about 9:1 (such as about 9:1 or about 8:1). In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising an mTOR inhibitor (such as rapamycin) associated (e.g., coated) with albumin (such as human albumin or human serum albumin), wherein the average diameter of the nanoparticles is about 150 nm, and wherein the weight ratio of albumin to mTOR inhibitor in the composition is not greater than about 9:1 (such as about 9:1 or about 8:1). In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising rapamycin associated (e.g., coated) with human albumin (such as human serum albumin), wherein the average diameter of the nanoparticles is not greater than about 150 nm (e.g., about 100 nm), and wherein the weight ratio of albumin to rapamycin in the composition is about 9:1 or about 8:1. In some embodiments, the average or mean diameter of the nanoparticles is from about 10 nm to about 150 nm. In some embodiments, the average or mean diameter of the nanoparticles is from about 40 nm to about 120 nm. In some embodiments, the average or mean diameter of the nanoparticles is about 100 - 120 nm, such as about 100 nm.

[0159] In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising an mTOR inhibitor (such as rapamycin) associated (e.g., coated) with albumin (such as human albumin or human serum albumin), wherein the composition further includes a sugar, and wherein the weight ratio of albumin to mTOR inhibitor in the composition is not greater than about 9:1 (such as about 9:1 or about 8:1). In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising an mTOR inhibitor (such as rapamycin) associated (e.g., coated) with albumin (such as human albumin or human serum albumin), wherein the composition further includes a sugar, wherein the average diameter of the nanoparticles is not greater than about 200 nm, and wherein the weight ratio of albumin to mTOR inhibitor in the composition is not greater than about 9:1 (such as about 9:1 or about 8:1). In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising an mTOR inhibitor (such as rapamycin) associated (e.g., coated) with albumin (such as human albumin or human serum albumin), wherein the composition further includes a sugar, wherein the average diameter of the nanoparticles is not greater than about 150 nm, and wherein the weight ratio of albumin to mTOR inhibitor in the composition is not greater than about 9:1 (such as about 9:1 or about 8:1). In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising an mTOR inhibitor (such as rapamycin) associated (e.g., coated) with albumin (such as human albumin or human serum albumin), wherein the composition further includes a sugar, wherein the average diameter of the nanoparticles is about 150 nm, and wherein the weight ratio of albumin to mTOR inhibitor in the composition is not greater than about 9:1 (such as about 9:1 or about 8:1). In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising rapamycin associated (e.g., coated) with human albumin (such as human serum albumin), wherein the composition further includes a sugar, wherein the average diameter of the nanoparticles is not greater than about 150 nm (e.g., about 100 nm), and wherein the weight ratio of albumin to rapamycin in the composition is about 9:1 or about 8:1. In some embodiments, the average or mean diameter of the nanoparticles is from about 10 nm to about 150 nm. In some embodiments, the average or mean diameter of the nanoparticles is from about 40 nm to about 120 nm. In some embodiments, the average or mean diameter of the nanoparticles is about 100 - 120 nm, such as about 100 nm.

[0160] In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising an mTOR inhibitor (such as rapamycin) stabilized by albumin (such as human albumin or human serum albumin). In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising an mTOR inhibitor (such as rapamycin) stabilized by albumin (such as human albumin or human serum albumin), wherein the average diameter of the nanoparticles is not greater than about 200 nm. In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising an mTOR inhibitor (such as rapamycin) stabilized by albumin (such as human albumin or human serum albumin), wherein the average diameter of the nanoparticles is not greater than about 150 nm. In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising an mTOR inhibitor (such as rapamycin) stabilized by albumin (such as human albumin or human serum albumin), wherein the average diameter of the nanoparticles is not greater than about 150 nm (e.g., about 100 nm). In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising rapamycin stabilized by human albumin (such as human serum albumin), wherein the average diameter of the nanoparticles is not greater than about 150 nm (e.g., about 100 nm). In some embodiments, the average or mean diameter of the nanoparticles is from about 10 nm to about 150 nm. In some embodiments, the average or mean diameter of the nanoparticles is from about 40 nm to about 120 nm. In some embodiments, the average or mean diameter of the nanoparticles is about 100 - 120 nm, such as about 100 nm.

[0161] In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising an mTOR inhibitor (such as rapamycin) stabilized by albumin (such as human albumin or human serum albumin), wherein the composition further includes a sugar. In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising an mTOR inhibitor (such as rapamycin) stabilized by albumin (such as human albumin or human serum albumin), wherein the composition further includes a sugar, wherein the average diameter of the nanoparticles is no greater than about 200 nm. In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising an mTOR inhibitor (such as rapamycin) stabilized by albumin (such as human albumin or human serum albumin), wherein the composition further includes a sugar, wherein the average diameter of the nanoparticles is no greater than about 150 nm. In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising an mTOR inhibitor (such as rapamycin) stabilized by albumin (such as human albumin or human serum albumin), wherein the composition further includes a sugar, wherein the average diameter of the nanoparticles is no greater than about 150 nm (e.g., about 100 nm). In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising rapamycin stabilized by human albumin (such as human serum albumin), wherein the composition further includes a sugar, wherein the average diameter of the nanoparticles is no greater than about 150 nm (e.g., about 100 nm). In some embodiments, the average or mean diameter of the nanoparticles is from about 10 nm to about 150 nm. In some embodiments, the average or mean diameter of the nanoparticles is from about 40 nm to about 120 nm. In some embodiments, the average or mean diameter of the nanoparticles is about 100 - 120 nm, such as about 100 nm.

[0162] In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising an mTOR inhibitor (such as rapamycin) stabilized by albumin (such as human albumin or human serum albumin), wherein the weight ratio of albumin to mTOR inhibitor in the composition is not greater than about 9:1 (such as about 9:1 or about 8:1). In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising an mTOR inhibitor (such as rapamycin) stabilized by albumin (such as human albumin or human serum albumin), wherein the average diameter of the nanoparticles is not greater than about 200 nm, and wherein the weight ratio of albumin to mTOR inhibitor in the composition is not greater than about 9:1 (such as about 9:1 or about 8:1). In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising an mTOR inhibitor (such as rapamycin) stabilized by albumin (such as human albumin or human serum albumin), wherein the average diameter of the nanoparticles is not greater than about 150 nm, and wherein the weight ratio of albumin to mTOR inhibitor in the composition is not greater than about 9:1 (such as about 9:1 or about 8:1). In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising an mTOR inhibitor (such as rapamycin) stabilized by albumin (such as human albumin or human serum albumin), wherein the average diameter of the nanoparticles is about 150 nm, and wherein the weight ratio of albumin to mTOR inhibitor in the composition is not greater than about 9:1 (such as about 9:1 or about 8:1). In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising rapamycin stabilized by human albumin (such as human serum albumin), wherein the average diameter of the nanoparticles is not greater than about 150 nm (e.g., about 100 nm), and wherein the weight ratio of albumin to rapamycin in the composition is about 9:1 or about 8:1. In some embodiments, the average or mean diameter of the nanoparticles is from about 10 nm to about 150 nm. In some embodiments, the average or mean diameter of the nanoparticles is from about 40 nm to about 120 nm. In some embodiments, the average or mean diameter of the nanoparticles is about 100 - 120 nm, such as about 100 nm.

[0163] In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising an mTOR inhibitor (such as rapamycin) stabilized by albumin (such as human albumin or human serum albumin), wherein the composition further includes a sugar, and wherein the weight ratio of albumin to mTOR inhibitor in the composition is not greater than about 9:1 (such as about 9:1 or about 8:1). In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising an mTOR inhibitor (such as rapamycin) stabilized by albumin (such as human albumin or human serum albumin), wherein the composition further includes a sugar, wherein the average diameter of the nanoparticles is not greater than about 200 nm, and wherein the weight ratio of albumin to mTOR inhibitor in the composition is not greater than about 9:1 (such as about 9:1 or about 8:1). In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising an mTOR inhibitor (such as rapamycin) stabilized by albumin (such as human albumin or human serum albumin), wherein the composition further includes a sugar, wherein the average diameter of the nanoparticles is not greater than about 150 nm, and wherein the weight ratio of albumin to mTOR inhibitor in the composition is not greater than about 9:1 (such as about 9:1 or about 8:1). In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising an mTOR inhibitor (such as rapamycin) stabilized by albumin (such as human albumin or human serum albumin), wherein the composition further includes a sugar, wherein the average diameter of the nanoparticles is about 150 nm, and wherein the weight ratio of albumin to mTOR inhibitor in the composition is not greater than about 9:1 (such as about 9:1 or about 8:1). In some embodiments, the mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising rapamycin stabilized by human albumin (such as human serum albumin), wherein the composition further includes a sugar, wherein the average diameter of the nanoparticles is not greater than about 150 nm (e.g., about 100 nm), and wherein the weight ratio of albumin to rapamycin in the composition is about 9:1 or about 8:1. In some embodiments, the average or mean diameter of the nanoparticles is from about 10 nm to about 150 nm. In some embodiments, the average or mean diameter of the nanoparticles is from about 40 nm to about 120 nm. In some embodiments, the average or mean diameter of the nanoparticles is about 100 - 120 nm, such as about 100 nm.

[0164] In some embodiments, the mTOR inhibitor nanoparticle composition comprises nab-rapamycin. In some embodiments, the mTOR inhibitor nanoparticle composition is nab-rapamycin. Nab-rapamycin is a formulation of rapamycin stabilized by human albumin USP and can be dispersed in a directly injectable physiological solution. The weight ratio of human albumin to rapamycin is from about 8:1 to about 9:1. When dispersed in a suitable aqueous medium such as 0.9% sodium chloride injection or 5% dextrose injection, nab-rapamycin forms a stable colloidal suspension of rapamycin. The mean particle size of the nanoparticles in the colloidal suspension is about 100 nanometers. Because HAS is soluble in water, nab-rapamycin can be reconstituted over a wide concentration range from a dilute solution (0.1 mg / ml rapamycin or its derivative) to a concentrated solution (20 mg / ml rapamycin or its derivative), including for example from about 2 mg / ml to about 8 mg / ml, or about 5 mg / ml.

[0165] Methods for preparing nanoparticle compositions are known in the art. For example, nanoparticles containing an mTOR inhibitor such as rapamycin and albumin such as human serum albumin or human albumin can be prepared under conditions of high shear force (e.g., sonication, high pressure homogenization, etc.). These methods are disclosed in, for example, U.S. Patent Nos. 5,916,596; 6,506,405; 6,749,868, 6,537,579, 7,820,788 and 8,911,786, and U.S. Patent Publication Nos. 2007 / 0082838, 2006 / 0263434 and PCT Application WO08 / 137148.

[0166] Briefly, the mTOR inhibitor such as rapamycin is dissolved in an organic solvent and the solution can be added to the albumin solution. The mixture is subjected to high pressure homogenization. The organic solvent can then be removed by evaporation. The resulting dispersion can be further lyophilized. Suitable organic solvents include, for example, ketones, esters, ethers, chlorinated solvents, and other solvents known in the art. For example, the organic solvent can be dichloromethane or chloroform / ethanol (e.g., in a ratio of 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1 or 9:1).

[0167] Other components in the mTOR inhibitor nanoparticle composition

[0168] The nanoparticles described herein can be present in a composition comprising other reagents, carriers, excipients, diluents, or stabilizers. For example, to increase the stability by increasing the negative ζ potential of the nanoparticles, certain negatively charged components can be added. Such negatively charged components include, but are not limited to, bile salts of bile acids consisting of bile acids such as cholic acid, chenodeoxycholic acid, taurocholic acid, glycodeoxycholic acid, taurochenodeoxycholic acid, lithocholic acid, ursodeoxycholic acid, dehydrocholic acid, etc.; phospholipids, which include lecithin (egg yolk)-based phospholipids, which include phosphatidylcholines such as palmitoyl oleoyl phosphatidylcholine, palmitoyl linoleoyl phosphatidylcholine, stearoyl linoleoyl phosphatidylcholine, stearoyl oleoyl phosphatidylcholine, stearoyl arachidoyl phosphatidylcholine, and dipalmitoyl phosphatidylcholine. Other phospholipids include L-α-dimyristoyl phosphatidylcholine (DMPC), dioleoyl phosphatidylcholine (DOPC), distearoyl phosphatidylcholine (DSPC), hydrogenated soy lecithin (HSPC), and other related compounds. Negatively charged surfactants or emulsifiers are also suitable as additives, for example, sodium cholesteryl sulfate, etc.

[0169] In some embodiments, the composition is suitable for administration to a human. In some embodiments, the composition is suitable for administration to a mammal such as domestic pets and agricultural animals in a veterinary context. In some embodiments, the composition is suitable for administration after reconstitution.

[0170] Examples of suitable carriers, excipients, and diluents include, but are not limited to, lactose, glucose, sucrose, sorbitol, mannitol, starch, gum arabic, calcium phosphate, alginate, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, saline solution, syrup, methylcellulose, methyl hydroxybenzoate and propyl hydroxybenzoate, talc, magnesium stearate, and mineral oil. The formulation can additionally include lubricants, wetting agents, emulsifiers, and suspending agents and / or preservatives.

[0171] Formulations suitable for subcutaneous administration include aqueous and non-aqueous isotonic sterile injection solutions, which can contain antioxidants, buffers, bacteriostatic agents, and solutes that render the formulation compatible with the blood of the intended recipient, and aqueous and non-aqueous sterile suspensions that can include suspending agents, solubilizers, thickening agents, stabilizers, and preservatives. The formulation can be present in unit dose or multi-dose sealed containers such as ampoules and vials, and can be stored under lyophilized (freeze-dried) conditions and only require the addition of a sterile liquid excipient such as water before use. The temporary solutions and suspensions can be prepared from sterile powders, granules, and tablets of the aforementioned types.

[0172] In some embodiments, the composition is formulated to have a pH range of from about 4.5 to about 9.0, including, for example, a pH range of from about 5.0 to about 8.0, from about 6.5 to about 7.5, and from about 6.5 to about 7.0. In some embodiments, the pH of the composition is formulated to be not less than about 6, including, for example, not less than about 6.5, 7, or 8 (such as about 8). The composition may also be made isotonic with blood by adding a suitable tonicity modifier such as glycerol.

[0173] Kit

[0174] In some embodiments, kits are provided that can be used for various purposes, such as, for example, treating a disease in an individual. The kits of the present invention include one or more containers that include an mTOR inhibitor nanoparticle composition (such as a rapamycin / albumin nanoparticle composition) (or unit dosage form and / or article) suitable for subcutaneous administration, and in some embodiments, further include a device for subcutaneous administration of the mTOR inhibitor nanoparticle composition. In some embodiments, the kit further includes instructions for use according to any of the methods described herein. The kit may further include a description of the individual selected for treatment. The instructions provided in the kits of the present invention are typically written instructions on a label or package insert (e.g., a sheet of paper included in the kit), but machine-readable instructions (e.g., instructions carried on a disk or optical storage disk) are also acceptable.

[0175] The kits of the present invention are packaged in a suitable package. Suitable packages include, but are not limited to, vials, bottles, jars, flexible packages (e.g., sealed or plastic bags), etc. The kit may optionally provide additional components, such as buffers and explanatory information. Accordingly, the present application also provides articles that include vials (e.g., sealed vials), bottles, jars, flexible packages, etc.

[0176] Instructions related to the use of mTOR inhibitor nanoparticle compositions typically include information on dosage, dosing regimens, and routes of administration for the intended treatment. The container can be a unit dose, bulk packaging (e.g., multi-dose packaging), or sub-unit dose. For example, a kit can be provided that contains a sufficient dose of an mTOR inhibitor nanoparticle composition (such as a rapamycin / albumin nanoparticle composition) to provide effective treatment to an individual over an extended period of time, such as any one of 1 week, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 2 weeks, 3 weeks, 4 weeks, 6 weeks, 8 weeks, 3 months, 4 months, 5 months, 7 months, 8 months, 9 months, or longer. The kit can also include a multi-unit dose of an mTOR inhibitor nanoparticle composition (such as a rapamycin / albumin nanoparticle composition) and instructions for use, packaged in an amount sufficient for storage and use in a pharmacy (e.g., a hospital pharmacy and a compounding pharmacy).

[0177] The kit can further include a device containing the mTOR inhibitor nanoparticle composition. The instructions can further include instructions for use of the device.

[0178] Examples

[0179] The present application can be better understood by reference to the following non-limiting examples, which are provided as exemplary embodiments of the present application. The following examples are provided to more fully illustrate the embodiments, but should in no way be construed as limiting the broad scope of the present application. Although certain embodiments of the present application have been shown and described herein, it is obvious that these embodiments are provided by way of example only. Various variations, changes, and substitutions can be envisioned by those skilled in the art without departing from the spirit and scope of the invention. It should be understood that various alternatives to the embodiments described herein can be employed in practicing the methods described herein.

[0180] Example 1: Pharmacokinetic Study of ABI-009 after Subcutaneous and Intravenous Administration in Sprague Dawley (SD) Rats

[0181] Female SD rats received a single dose of nab-rapamycin (ABI-009) subcutaneously (i.e., "SC" or "subQ") or intravenously (IV). The study design is summarized in Table 1 below. No inflammation or toxicity was observed at any time point after administration at the subcutaneous injection site compared to the saline control (vehicle).

[0182]

[0183] After subcutaneous or intravenous injection of ABI-009, the concentration of rapamycin in whole blood was measured at different time points. The results of whole blood collection are summarized in Tables 2 and 3 below.

[0184]

[0185]

[0186]

[0187] Surprisingly, as summarized in Table 4 below, subcutaneous administration enhanced bioavailability compared to intravenous administration, as represented by the total area under the curve (AUC). Subcutaneous administration of only 0.56 mg / kg of ABI-009 produced a drug exposure similar to 1 / 3 of the dose of IV ABI-009 (1.7 mg / kg). Further, subcutaneous administration decreased the maximum concentration reached (C 最大 ), and delayed the time to reach the maximum concentration (C 最长时间 ). Higher subcutaneous ABI-009 doses increased the peak level and AUC of rapamycin in the blood.

[0188]

[0189]

[0190] Example 2: Biodistribution of ABI-009 after administration in rats

[0191] Tissues were harvested from the rats described in Example 1 above 24 hours or 168 hours after subcutaneous (subQ) or intravenous (IV) administration of ABI-009 (see Table 1 for study design). In Figure 5 (bone marrow and brain), Figure 6 (heart and lung), and Figure 7 (lung and pancreas), the concentrations of rapamycin in specific rat tissues were indicated at 24 hours or 168 hours after administration.

[0192] The subcutaneous administration route resulted in significant distribution in all tested organs, including bone marrow, brain, heart, liver, lung, and pancreas. The patterns of subcutaneous and intravenous organ distribution were similar, but subcutaneous administration at a dose of 0.56 mg / kg was able to produce tissue concentrations similar to those of intravenous administration at a dose of 1.7 mg / kg. In well-perfused organs, including the heart, liver, lung, and pancreas, the rapamycin concentration decreased significantly between 24 hours and 168 hours. However, the concentration in the brain was relatively stable between 24 and 168 hours.

[0193] To further clarify the differences between the brain and blood distribution of rapamycin, further experiments were conducted on rats. A single dose of nab-rapamycin (ABI-009) was administered subcutaneously to rats at a dose of 1.7 mg / kg, 9.5 mg / kg, or 17 mg / kg. Rats were sacrificed at 24, 72, and 120 hours and whole blood and brain tissues were collected. Rapamycin concentrations were measured at each time point for each sample. As Figure 5 indicated, a dose-dependent increase in brain rapamycin levels was observed. Surprisingly, while the blood levels of rapamycin rapidly approached baseline, even at the high dose of 17 mg / kg, the brain rapamycin levels remained good throughout the 120 hours, even at the lowest dose. See also Figure 8 .

[0194] Example 3: Nab-rapamycin nanoparticle formulations containing sugars

[0195] Nab-rapamycin (ABI-009) formulations with and without sugars will be prepared, including sucrose formulations and trehalose formulations. The formulations will be lyophilized and then reconstituted with water at various concentrations of rapamycin from 1 mg / ml to 40 mg / ml. The formulations will then be lyophilized again and incubated at 40 °C for 15 days.

[0196] After incubation, the formulations will be reconstituted with water and analyzed for albumin oligomers and polymers and reconstitution time simultaneously or subsequently.

[0197] Formulations showing reduced albumin oligomers and polymers and / or rapid reconstitution will be selected as enhanced formulations for subcutaneous administration.

[0198] Example 4: Toxicology study after repeated subcutaneous administration of ABI-009 in SD rats

[0199] The purpose of the study was to evaluate the overall safety and local toxicity at the injection site after repeated ABI-009 SC injections in SD rats. Signs of clinical distress were observed to determine toxicity. Signs of inflammation and necrosis in skin samples from the injection site were analyzed by histopathology.

[0200] The study used 15 female Sprague Dawley (SD) rats weighing 160 - 180 g. ABI-009 was dissolved in saline to prepare a stock solution (10 mg / ml), and then further diluted in 0.9% saline solution of HSA to prepare for subcutaneous injection (volume: 1.0 ml / kg).

[0201] A. Study design

[0202] The rats were divided into 5 groups of 3 animals each. As shown in Table 5, the rats were weighed and administered subcutaneously once every 4 days for 4 weeks (7 injections).

[0203]

[0204] SC = Subcutaneous injection

[0205] Check the clinical signs of the overall toxicity of the animals daily and check the reaction of the local injection site to subcutaneous injection.

[0206] Collect whole blood samples before each injection for the animals receiving ABI-009 (Groups 3, 4, and 5) and analyze the rapamycin trough concentration.

[0207] All animals were euthanized after 4 weeks, and the local toxicity signs of skin samples from the local injection site were examined by histopathology.

[0208] B. Experimental procedures

[0209] 1. Preparation of dosing solutions

[0210] The vehicle controls included 0.9% saline solution and HAS in 0.9% saline solution. Based on a 9:1 albumin:rapamycin ratio of the test article ABI-009 (manufacturing batch #C345-001, Fisher batch #51394.2), the final concentration of the HSA solution was 90 mg / ml. Each vial of ABI-009 (C345-001) contained 97.4 mg rapamycin and 874 mg human albumin. The HSA saline solution was diluted from a 20% Grifols albumin stock solution (200 mg / ml).

[0211] For the ABI-009 dosing solution, first prepare a 10 mg / ml ABI-009 stock solution, and then dilute the dosing solution to the desired concentration using the HSA-saline solution. Dissolve 1 vial of 100 mg of ABI-009 in 10 ml of 0.9% saline to prepare a 10 mg / ml solution.

[0212] Prepare a 5 mg / ml ABI-009 solution by diluting 0.6 ml of the stock solution (10 mg / ml) with 0.6 ml of HSA-0.9% saline to prepare a 5.0 mg / ml solution for Group 4. Prepare a 1.7 mg / ml ABI-009 solution by diluting 0.3 ml of the ABI-009 solution for Group 4 (5.0 mg / ml) with 0.6 ml of HSA-0.9% saline to prepare a 1.7 mg / ml solution for Group 3.

[0213] 2. Administration

[0214] The rats were anesthetized, weighed, and administered ABI-009 solution, HSA solution, and saline by subcutaneous injection (SC) according to Table 6, once every 4 days for 4 weeks (7 injections).

[0215]

[0216]

[0217] The rats were examined once a day for clinical signs of total toxicity and the response of the local injection site to subcutaneous injection. Signs of clinical distress were observed to determine toxicity. Ruffled fur, weight loss, lethargy, discharge, neurological symptoms, morbidity, redness and inflammation at the injection site, and any other signs considered abnormal in the behavior of the animals. Photos of the injection sites of all rats were taken before and after SC injection.

[0218] 3. Sample Collection and Analysis

[0219] For the rats treated with ABI-009 (Groups 3, 4, and 5), the rats were anesthetized and bled to place samples in pre-chilled K2EDTA tubes before each administration (except for the first dose). Whole blood was collected, stored in labeled Eppendorf tubes at -80 °C, and analyzed for rapamycin trough concentration.

[0220] All animals were euthanized at the final euthanasia point on Day 29 (4 weeks, 96 hours after administration of ABI-009 on Day 25). At the final euthanasia time point, whole blood samples were collected to analyze rapamycin trough levels. The brain, lungs, liver, heart, pancreas, and bone marrow were collected, rinsed with saline to remove blood, divided into two parts, snap-frozen in separate labeled tubes, and stored at -80 °C. The frozen blood samples from the ABI-009 treatment groups (Groups 3, 4, and 5) were shipped on dry ice to BASi. The rapamycin trough blood concentration was analyzed by BASi using the LC / MS / MS method.

[0221] At the final euthanasia time point, the skin and subcutis of the SC administration area were excised for histological analysis by H&E staining to analyze signs of inflammation by histopathology. Fifteen formalin-fixed rat skin samples were histopathologically measured and routinely processed. One section of each block was cut and stained with hematoxylin and eosin (H&E). The sections were evaluated by board-certified veterinary pathologists using an optical microscope. The severity grade of histological lesions was 0 - 5 (0 = absent / normal, 1 = mildest, 2 = mild, 3 = moderate, 4 = marked, 5 = severe). The mean scores of different groups were analyzed by t-test.

[0222] C. Results

[0223] 1. Systemic toxicity

[0224] Clinical signs of distress were observed daily to determine toxicity. These included piloerection, weight loss, lethargy, morbidity, neurological signs, redness and inflammation at the injection site, and any other signs considered abnormal in the behavior of the animals. At the current dose schedule (1.7 - 10 mg / kg, 7 doses), rats were normal after administration of saline, HSA, and ABI - 009, and no clinical signs of stress were observed during the study.

[0225] There was no weight loss (<20%), and all treatment groups gained weight during the study (Table 7). The results indicate that rats tolerated subcutaneous injection of ABI - 009 in the dose range of 1.7 - 10.0 mg / kg.

[0226]

[0227] 2. Local toxicity

[0228] Histopathological measurements were made on 15 formalin - fixed rat skin samples from the SC administration area. Histopathological findings in the skin samples included inflammatory cell necrosis and mixed infiltration in the perivascular area; both lesions were observed in the subcutaneous tissue / hypodermis.

[0229] Necrosis was focal and characterized by loss of normal cells, neutrophil infiltration, hemorrhage, and fibrin exudation, with variable adjacent fibrinous hyperplasia. Necrosis was observed only in samples from animals treated with ABI - 009 at dose levels of 5 mg / kg (Group 4, 1 animal with minimal necrosis) and 10 mg / kg (Group 5, all 3 animals with mild to marked necrosis), while saline (Group 1), HSA (Group 2), and ABI - 009 at 1.7 mg / kg (Group 3) did not cause necrosis. See Table 10 and Figure 14 . Compared to the HSA group, only the highest dose of ABI - 009 at 10 mg / kg showed a significantly increased necrosis score (P = 0.02, t - test).

[0230]

[0231]

[0232] The mixed inflammatory cell infiltrate in the subcutaneous perivascular area was characterized by infiltration and accumulation of lymphocytes, plasma cells, macrophages, occasional multinucleated giant cells, and variable numbers of neutrophils. Mixed inflammatory cell infiltrates were observed in all treatment groups, with the highest mean scores in animals treated with HSA (Group 2) and ABI-009 at 10 mg / kg (Group 5). For the low-dose ABI-009 injection of 1.7 mg / kg (Group 3), the mean score was similar to the control group receiving saline injection (Group 1). See Tables 8 and Figure 9 The high mixed inflammatory cell infiltration observed in the HSA group (Group 2) compared with the saline control (P = 0.01, t-test) indicated that the local inflammation was mainly caused by the injection of the foreign protein human serum albumin.

[0233] exist Figures 10 - 14 Representative histological images of rats in each group are shown in .

[0234] For the ABI-009 treatment group, there was a dose-related increase in local toxicity with increasing ABI-009 doses. At the lowest dose of ABI-009 at 1.7 mg / kg, the histology of the local injection site was similar to that of the saline control group; while in animals treated with ABI-009 at the 10 mg / kg dose level, necrosis and subcutaneous inflammatory cell infiltration were most severe.

[0235] 3. Rapamycin trough blood levels

[0236] Rapamycin trough blood samples were collected for the groups treated with ABI-009 (except for the 1st dose on day 1) before each injection (days 5, 9, 13, 17, 21, 25, 29) and analyzed by BASi using the LC / MS / MS method. The trough levels are shown in Table 9. Four days after SC injection, most rapamycin trough blood levels were consistently in the range of 2-20 ng / ml. Two samples in the ABI-009 10 mg / kg group (Group 5) were clearly outliers. The reason for this observation could not be determined. However, the abnormally high trough levels occurred only in the highest ABI-009 dose group, which also showed mild to significant necrosis in the subcutaneous tissue, suggesting that the skin lesions may hinder the normal absorption of ABI-009 and lead to prolonged drug retention.

[0237]

[0238] For each ABI-009 treatment group, there was no significant drug accumulation during the study as the rapamycin trough blood levels generally remained stable. As the ABI-009 dose increased, the mean trough concentration of rapamycin increased in a dose-dependent manner. Higher trough levels were observed in the groups receiving 5 mg / kg (P = 0.06) and 10 mg / kg (P = 0.01) of ABI-009 compared to the 1.7 mg / kg ABI-009 group( Figure 15 ).

[0239] In summary, at the current dosing regimen (1.7 - 10 mg / kg, 7 doses), rats were normal after ABI-009 administration and no weight loss was observed during the study. Histopathological findings demonstrated dose-related signs of local toxicity with mild to marked necrosis at the highest ABI-009 dose (10 mg / kg). The mixed inflammatory cell infiltration may be caused by the foreign protein HSA. ABI-009 at 1.7 mg / kg (solution concentration 1.7 mg / ml) showed a local injection response similar to that of the saline control. There was no significant drug accumulation after repeated SC injections. As the ABI-009 dose increased, the rapamycin trough blood levels increased.

[0240] The results showed that rats were systemically tolerant to multiple doses of ABI-009 in the range of 1.7 - 10.0 mg / kg by subcutaneous injection. Locally, they were well-tolerant to the ABI-009 solution at a concentration of 1.7 mg / ml. No adverse effects were observed at this dose level.

[0241] Example 5: Antitumor Activity Study of nab-Rapamycin

[0242] A study was conducted to compare the antitumor activity of rapamycin (Rapamune) by the oral route and nab-rapamycin (ABI-009) by the intravenous or subcutaneous route in a human hepatocellular carcinoma xenograft mouse model.

[0243] Human cancer cells for mouse injection were prepared by thawing (using liquid nitrogen) frozen SNU-398 (TSC2-deficient human hepatocellular carcinoma cells) obtained from (CRL-2233 TM ). The cells were dispersed into a 75 cm 2 flask containing RPMI 1640 medium supplemented with 10% fetal bovine serum and incubated at 37 °C in a humidified 5% CO2 atmosphere. At 80% cell confluence, the cells were expanded into a 150 cm 2 flask with fresh medium. The cells were allowed to grow to obtain 1 × 10 7 (2 × 10 7) targets of cells.

[0244] Twenty athymic nude mice were housed in filter - topped cages. In 0.1 ml of phosphate - buffered saline containing 20% the cancer cells were subcutaneously injected into both flanks (1×10 7 ) per flank.

[0245] Tumors began to appear on day 1 of treatment (average tumor size ∼100 - 150 mm 3 ). The animals were divided into 4 groups.

[0246] Group 1, consisting of 5 mice, received saline via the intravenous route, twice a week for 6 weeks.

[0247] Group 2, consisting of 5 mice, received ABI - 009 at 7.5 mg / kg via the intravenous route, twice a week for 6 weeks. The total dose of rapamycin was 15 mg / kg / wk.

[0248] Group 3, consisting of 5 mice, received rapamycin at 3 mg / kg via oral administration, five times a week for 6 weeks. The total dose of rapamycin was 15 mg / kg / wk.

[0249] Group 4, consisting of 3 mice, received ABI - 009 at 7.5 mg / kg via the subcutaneous route, twice a week for 6 weeks. The total dose of rapamycin was 15 mg / kg / wk.

[0250] Measurements (mouse body weight and tumor measurements) were taken three times a week (Monday, Wednesday, and Friday) until the scheduled sacrifice time point and terminated after 6 weeks or when the tumor reached a maximum volume of 2,000 mm 3 . Signs of distress were recorded daily. Tumors were harvested and stored. Blood samples were collected at the same time as the tumors were collected.

[0251] Result : The study is in progress. The initial tumor volume results (mean and standard error of the mean, SEM) for each group are summarized in Table 10 below. The tumor growth inhibition (TGI) compared to saline (Group 1) and the P - value of TGI relative to saline are also reported in Table 10. The results are also summarized in Figure 16 .

[0252]

[0253] Compared with the saline control group, oral rapamycin solution at 15 mg / kg / wk (Group 3) resulted in moderate tumor growth inhibition (TGI 33.2%, P = not significant). Intravenous administration of equidose ABI-009 weekly (Group 2) resulted in significantly higher TGI than oral rapamycin (TGI 66.7% relative to the saline control group, P = 0.0016 relative to oral rapamycin). However, ABI-009 administered via the subcutaneous route (Group 4) produced the most significant tumor growth inhibition (TGI 89.8%, P = 0.0001 relative to the saline control group, P < 0.0001 relative to oral rapamycin).

[0254] No signs of toxicity were observed in any treatment group. No significant weight loss (>10%) was observed in any treatment group. By day 15, a slight weight loss was observed in the saline control group (Group 1), while each treatment group (Groups 2-4) maintained or gained weight by day 15. The weight results are summarized in Figure 17 in.

[0255] In summary, in the TSC2-deficient SNU-398 human hepatocellular carcinoma xenograft mouse model, ABI-009 administered via the intravenous or subcutaneous route produced significantly higher anti-tumor activity compared to equidose oral rapamycin weekly. Even compared to ABI-009 via the intravenous route, ABI-009 via the subcutaneous route was unexpectedly effective. No major toxicity or weight loss was observed in any treatment group.

Claims

1. A method of treating a disease in an individual, comprising subcutaneously administering to the individual a pharmaceutical composition comprising nanoparticles comprising an mTOR inhibitor and albumin, wherein the amount of the mTOR inhibitor in the pharmaceutical composition is about 0.1 mg / m 2 to about 10 mg / m 2 per dose.

2. The method according to claim 1, wherein the amount of the mTOR inhibitor in the pharmaceutical composition is about 1 mg / m 2 to about 10 mg / m 2 dose.

3. The method according to claim 1 or claim 2, wherein the amount of the mTOR inhibitor in the pharmaceutical composition is about 5 mg / m 2 dose.

4. The method according to any one of claims 1 - 3, wherein the pharmaceutical composition further comprises sugar.

5. The method according to any one of claims 1 - 4, wherein the pharmaceutical composition is administered once a week or less.

6. The method according to claim 5, wherein the pharmaceutical composition is administered once a week.

7. The method according to claim 5, wherein the pharmaceutical composition is administered twice every three weeks.

8. The method according to any one of claims 1 - 7, wherein the disease is cancer.

9. The method according to any one of claims 1 - 7, wherein the disease is a mitochondrial disease.

10. The method according to any one of claims 1 - 9, wherein the individual is a human.

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