HK40003230A - Intravascular delivery of nanoparticle compositions and uses thereof - Google Patents

Description

Intravascular delivery of nanoparticle compositions and uses thereof

The present application is a divisional application filed on 2012/4/27/application No. 201280032010.2 entitled "intravascular delivery of nanoparticle compositions and uses thereof".

RELATED APPLICATIONS

This application claims the benefit of priority from U.S. provisional application serial No. 61/518,084 filed on 28/2011 and U.S. provisional application serial No. 61/557,851 filed on 9/2011, which are incorporated herein in their entirety.

Technical Field

The present invention relates to methods of delivering and applying compositions comprising nanoparticles comprising a macrolide and an albumin by directly injecting the nanoparticle composition into the blood vessel wall or tissue surrounding the blood vessel wall.

Background

Coronary artery disease is one of the leading causes of death worldwide. Although coronary artery bypass surgery is an effective treatment for atherosclerosis and arterial stenosis due to other causes, it is a highly invasive procedure and requires a large number of hospital stays and recovery times. Percutaneous Transluminal Coronary Angioplasty (PTCA), often referred to as balloon angioplasty, is less invasive, less invasive and significantly less costly than bypass surgery. The efficacy of balloon angioplasty has been significantly improved by the introduction of a stent implantation procedure involving the placement of a stent structure in an artery that has been treated by balloon angioplasty. Stents inhibit the abrupt reclosing of the artery and have some benefit in reducing subsequent restenosis caused by hyperplasia. Despite such improvements, patients who have undergone angioplasty procedures and subsequent stenting still experience a high incidence of restenosis resulting from hyperplasia. Implanting a stent coated with an antiproliferative drug can significantly reduce the incidence of hyperplasia.

Albumin-based nanoparticlesParticulate compositions have been developed as drug delivery systems for the delivery of substantially water-insoluble drugs, such as taxane. See, for example, U.S. Pat. nos. 5,916,596; 6,506,405, respectively; 6,749,868 and 6,537,579, 7,820,788 and 7,923,536. It is generally believed that albumin-based nanoparticles such asWhen introduced into the blood stream, will dissolve to form albumin-drug complexes. Such complexes utilize the natural properties of the protein, albumin, to transport and deliver substantially water-insoluble drugs to sites of disease, such as tumor sites. In addition, albumin-based nanoparticle technology offers the ability to increase drug solubility by avoiding the need for toxic chemicals, such as solvents, during administration, thus potentially increasing safety by eliminating solvent-related side effects.

The disclosures of all publications, patents, patent applications, and published patent applications cited herein are hereby incorporated by reference in their entirety.

Summary of The Invention

The present application provides, in some embodiments, methods of delivering a composition comprising nanoparticles comprising albumin and a macrolide to a blood vessel, wherein the method comprises injecting into the blood vessel wall or tissue surrounding the blood vessel wall an effective amount of a composition comprising nanoparticles comprising a macrolide and albumin. In some embodiments, there is provided a method of inhibiting negative remodeling in a blood vessel in an individual in need thereof, comprising injecting into the blood vessel wall or tissue surrounding the blood vessel wall an effective amount of a composition comprising nanoparticles comprising a macrolide and an albumin. In some embodiments, there is provided a method of inhibiting vascular fibrosis (such as medial fibrosis or adventitial fibrosis) in a blood vessel of an individual in need thereof, comprising injecting into the blood vessel wall or tissue surrounding the blood vessel wall an effective amount of a composition comprising nanoparticles comprising a macrolide and an albumin. In some embodiments, there is provided a method of reducing the proliferative index of a blood vessel in an individual in need thereof, comprising injecting into the blood vessel wall or tissue surrounding the blood vessel wall an effective amount of a composition comprising nanoparticles comprising a macrolide and an albumin. In some embodiments, there is provided a method of promoting positive remodeling in a blood vessel in an individual in need thereof, comprising injecting into the blood vessel wall or tissue surrounding the blood vessel wall an effective amount of a composition comprising nanoparticles comprising a macrolide and an albumin.

In some embodiments, the blood vessel is an artery, such as a coronary artery or a peripheral artery. In some embodiments, the artery is selected from the group consisting of a renal artery, a cerebral artery, a pulmonary artery, and a leg artery. In some embodiments, the blood vessel is a vein.

In some embodiments, the nanoparticle composition is injected into the vessel wall. In some embodiments, the nanoparticle composition is injected into the tissue surrounding the vessel wall. In some embodiments, the nanoparticle composition is injected into adventitial tissue.

In some embodiments, the injected dose of the nanoparticle composition is from about 0.001mg to about 100mg, including for example from about 0.05mg to about 5 mg. In some embodiments, the nanoparticle composition has an injection volume of about 0.01ml to about 50ml, including, for example, about 0.5ml to about 5 ml. In some embodiments, the nanoparticle composition is injected through a catheter with a needle, such as a deployable needle. In some embodiments, the nanoparticle composition is injected at least once a year. In some embodiments, the nanoparticle composition is injected only once.

In some embodiments, the nanoparticle composition is injected distal to the disease site. In some embodiments, the nanoparticle composition is injected proximal to the disease site. In some embodiments, the nanoparticle composition is injected at or near the site of the disease. In some embodiments, the nanoparticle composition is injected away from the disease site. In some embodiments, the nanoparticle composition is injected at least about 2cm (including, e.g., at least any of 3, 4, 5, 6,7, 8, 9, or 10cm) from the disease site.

In some embodiments according to any of the above embodiments, the subject has any of the following: angina, myocardial infarction, congestive heart failure, arrhythmia, peripheral artery disease, claudication, or chronic limb ischemia. In some embodiments, the individual is a human. In some embodiments, the method is performed during a vascular interventional procedure including, but not limited to, angioplasty (such as percutaneous transluminal coronary angioplasty), stent implantation, or atherectomy (atherectomy). In some embodiments, the method is performed after a vascular interventional procedure including, but not limited to, angioplasty, stenting, or atherectomy.

In some embodiments according to any of the above embodiments, the macrolide is rapamycin or a derivative thereof. In some embodiments, the macrolide is rapamycin. In some embodiments according to any of the preceding embodiments, the average diameter of the nanoparticles in the composition is no greater than about 200nm, such as no greater than about 100 nm. In some embodiments, the average diameter of the nanoparticles in the composition is not less than about 70 nm. In some embodiments, the macrolide in the nanoparticle is coated with albumin.

Also provided are kits and devices for use in any of the methods described herein. For example, in some embodiments, a catheter with a needle (e.g., a deployable needle) is provided, wherein the needle comprises a composition comprising nanoparticles comprising a macrolide and an albumin. In some embodiments, the macrolide is rapamycin. In some embodiments, the nanoparticle comprises a macrolide coated with albumin. In some embodiments, the average diameter of the nanoparticles in the composition is no greater than about 200nm, such as no greater than about 100 nm. In some embodiments, the average diameter of the nanoparticles in the composition is not less than about 70 nm.

These and other aspects and advantages of the present invention will become apparent from the following detailed description and appended claims. It is to be understood that one, some or all of the properties of the various embodiments described herein may be combined to form further embodiments of the invention.

Brief Description of Drawings

Figure 1 provides an image of a micro-infusion catheter for periadventitial injection of Nab-rapamycin for the femoral artery. FIG. 1A shows a deflated balloon that houses a needle. Fig. 1B shows the inflated balloon with the needles protruding outward.

FIG. 2 provides a flow chart of two study designs involving periadventitial injection of Nab-rapamycin in a porcine femoral balloon angioplasty injury model. Figure 2A shows a flow chart of a pharmacokinetic study. Figure 2B shows a histopathological study flow chart.

FIGS. 3A-F show a series of representative angiograms of periadventitial injections of Nab-rapamycin into the femoral artery.

Figures 4A-4D show the reduction of luminal stenosis following periadventitial delivery of Nab-rapamycin as measured by mean luminal cross-sectional area (4A), mean arterial cross-sectional area (4B), mean percent luminal stenosis (4C), and mean medial fibrosis (4D).

FIGS. 5A and 5B show the pharmacokinetics of Nab-rapamycin following periadventitial delivery in the femoral artery, as measured by serum rapamycin concentration (5A) and tissue rapamycin concentration (5B).

FIGS. 6A-6D show histopathological staining of femoral arteries treated with (6C and 6D) or (6A and 6B) with Nab-rapamycin by periadventitial delivery. Fig. 6A and 6C show staining with H & E. Fig. 6B and 6D show trichromatic staining.

FIG. 7A shows proliferation index following periadventitial delivery of Nab-rapamycin or a control. FIG. 7B shows endothelialization following periadventitial delivery of Nab-rapamycin or a control.

FIG. 8A shows proliferation indices at days 3, 8, and 28 following periadventitial delivery of Nab-rapamycin. FIG. 8B shows endothelialization on days 3, 8, and 28 following periadventitial delivery of Nab-rapamycin.

FIG. 9A shows adventitia leukocyte infiltration on days 3, 8, and 28 following periadventitial delivery of Nab-rapamycin or controls. FIG. 9B shows the mean number of adventitial vessels at day 28 after periadventitial delivery of Nab-rapamycin or control.

FIG. 10 shows re-endothelialization of target arteries at days 3, 8, and 28 following periadventitial delivery of Nab-rapamycin or controls.

Detailed Description

The present application provides methods of delivering a composition comprising nanoparticles comprising a macrolide and an albumin to a blood vessel ("nanoparticle composition"), wherein the method comprises injecting into the blood vessel wall or tissue surrounding the blood vessel wall an effective amount of a composition comprising nanoparticles comprising a macrolide and an albumin. The methods are useful, for example, for inhibiting negative remodeling of blood vessels and/or inhibiting vascular fibrosis of blood vessels, and thus, for treating a variety of diseases associated with negative remodeling and/or vascular fibrosis.

Using the porcine femoral artery balloon injury model, it was shown that a nanoparticle composition comprising a macrolide and albumin, namely nanoparticle albumin-bound (Nab) rapamycin (Nab-rapamycin), when injected into perivascular adventitial tissue, significantly reduced negative remodeling of balloon-injured vessels and intravascular-side fibrosis. Within one hour after injection, rapamycin levels in perivascular tissues were about 1500-fold higher than rapamycin levels in blood within one hour, with rapamycin remaining in perivascular tissues for at least 8 days. Thus, periadventitial injection of nanoparticle compositions can be an effective method of inhibiting negative remodeling, inhibiting vascular fibrosis, and treating a variety of diseases associated with negative remodeling and/or vascular fibrosis.

Accordingly, in one aspect, the present application provides a method of inhibiting negative remodeling in a blood vessel or fibrosis in a blood vessel in an individual in need thereof, comprising injecting into the blood vessel wall or tissue surrounding the blood vessel wall an effective amount of a composition comprising nanoparticles comprising a macrolide and an albumin.

In another aspect, there is provided a method of delivering a composition comprising nanoparticles comprising a macrolide and an albumin to a blood vessel, wherein the method comprises injecting into the blood vessel wall or tissue surrounding the blood vessel wall an effective amount of a composition comprising nanoparticles comprising a macrolide and an albumin.

Further provided are kits and devices (e.g., catheter with needle) that can be used in the methods described herein.

Definition of

The term "subject" refers to a mammal, including, but not limited to, a human, a cow, a horse, a cat, a dog, a rodent, or a primate.

It is to be understood that the aspects and embodiments of the invention described herein include "consisting of and/or" consisting essentially of aspects and embodiments.

A value or parameter of "about" herein refers to a variable that includes (and describes) the value or parameter itself. For example, a description of "about X" includes a description of "X".

As used herein and in the appended claims, the singular forms "a", "an" and "the" include plural referents unless the context clearly dictates otherwise.

Method of the invention

The present application provides, in some embodiments, methods of delivering a composition comprising nanoparticles comprising albumin and a macrolide (such as rapamycin or a derivative thereof, e.g., rapamycin) to a blood vessel, wherein the method comprises injecting into the blood vessel wall or tissue surrounding the blood vessel wall an effective amount of a composition comprising nanoparticles comprising a macrolide and albumin. In some embodiments, there is provided a method of delivering a composition comprising nanoparticles comprising albumin and a macrolide (such as rapamycin) to a blood vessel, wherein the method comprises injecting into the blood vessel wall or tissue surrounding the blood vessel wall an effective amount of a composition comprising nanoparticles comprising a macrolide and an albumin (such as human serum albumin), wherein the macrolide in the nanoparticles is coated with the albumin. In some embodiments, there is provided a method of delivering a composition comprising nanoparticles comprising albumin and a macrolide (such as rapamycin) to a blood vessel, wherein the method comprises injecting into the blood vessel wall or tissue surrounding the blood vessel wall an effective amount of a composition comprising nanoparticles comprising a macrolide and an albumin (such as human serum albumin), wherein the average particle size of the nanoparticles in the composition is no greater than about 200nm (such as less than about 200nm, for example no greater than about 100 nm). In some embodiments, there is provided a method of delivering a composition comprising nanoparticles comprising albumin and a macrolide (such as rapamycin) to a blood vessel, wherein the method comprises injecting into the blood vessel wall or tissue surrounding the blood vessel wall an effective amount of a composition comprising nanoparticles comprising rapamycin and albumin (such as human serum albumin), wherein the rapamycin in the nanoparticles is coated with the albumin, and wherein the average particle size of the nanoparticles in the composition is no greater than about 200nm (such as less than about 200nm, for example no greater than about 100 nm). In some embodiments, there is provided a method of delivering Nab-rapamycin to a blood vessel, wherein the method comprises injecting into the blood vessel wall or tissue surrounding the blood vessel wall an effective amount of Nab-rapamycin. In some embodiments, the nanoparticle composition is injected at or near the disease site (or injury site), such as no more than about 2, 1, or 0.5cm from the disease site (or injury site). In some embodiments, the nanoparticle composition is injected away from the disease site (e.g., at least about any of: 1, 2, 3, 4, 5, 6,7, 8, 9, or 10cm from the disease site).

The general vessel wall has an endothelium, which is the wall layer exposed to the lumen of the vessel. Beneath the endothelium is the basement membrane, which is in turn surrounded by the intima. The inner membrane is further surrounded by an inner elastic layer, on which the intermediate layer is located. Further, the intermediate layer is covered by an outer elastic layer, which acts as an outer barrier, separating the vessel wall from the adventitial tissue surrounding the vessel wall. The methods described herein include injecting a nanoparticle composition into any of these layers of the vessel wall. In some embodiments, the nanoparticle composition is infused into the endothelium. In some embodiments, the nanoparticle composition is impregnated into a substrate film. In some embodiments, the nanoparticle composition is injected into the inner membrane. In some embodiments, the nanoparticle composition is infused into the inner elastomeric layer. In some embodiments, the nanoparticles are infused into the intermediate layer. In some embodiments, the nanoparticles are impregnated into the outer elastomeric layer. In some embodiments, the nanoparticle composition is injected into any one of the following vascular regions: inner membrane (including endothelium, basement membrane, inner elastic layer), middle membrane (including smooth muscle cells), and outer membrane (including outer elastic membrane, collagen fibers).

"perivascular tissue" -used herein interchangeably with the terms "perivascular" or "periadventitial", refers to a region on the outer surface of a blood vessel wall. It includes adventitial tissue of the blood vessel, as well as regions beyond the adventitial tissue. By controlling the injection site of the nanoparticle composition, the nanoparticle composition can be injected to a specific desired location.

Methods and devices have been developed for the purpose of injecting therapeutic agents into the vessel wall and tissue surrounding the vessel wall. For example, catheters carrying needles capable of delivering therapeutic and other agents deep into the adventitial layer surrounding the lumen of a blood vessel have been described in U.S. Pat. Nos. 6,547,303, 6,860,867 and U.S. patent application publication Nos. 2007/0106257, 2010/0305546, and 2009/0142306, the entire contents of which are specifically incorporated herein by reference. The methods of the present invention in some embodiments utilize a catheter having a needle for nanoparticle composition injection. In some embodiments, the needle is deployable. The catheter may be advanced intravascularly to a target injection site (which may or may not be a disease region) in the vessel. A needle in the catheter is advanced through the vessel wall such that the hole on the needle is positioned in a desired area (e.g., perivascular area) and the nanoparticle composition can be injected through the needle hole into the desired area.

For example, in some embodiments, there is provided a method of delivering a composition comprising nanoparticles comprising albumin and a macrolide (such as rapamycin or a derivative thereof, e.g. rapamycin) to a blood vessel, wherein the method comprises injecting (e.g. via a catheter with a needle) into the tissue surrounding the blood vessel wall an effective amount of a composition comprising nanoparticles comprising a macrolide and albumin. In some embodiments, there is provided a method of delivering a composition comprising nanoparticles comprising albumin and a macrolide (such as rapamycin) to a blood vessel, wherein the method comprises injecting (e.g. via a catheter with a needle) into the tissue surrounding the blood vessel wall an effective amount of a composition comprising nanoparticles comprising a macrolide and an albumin (such as human serum albumin), wherein the macrolide in the nanoparticles is coated with the albumin. In some embodiments, there is provided a method of delivering a composition comprising nanoparticles comprising albumin and a macrolide (such as rapamycin) to a blood vessel, wherein the method comprises injecting (e.g. via a catheter with a needle) into the tissue surrounding the blood vessel wall an effective amount of a composition comprising nanoparticles comprising a macrolide and an albumin (such as human serum albumin), wherein the average particle size of the nanoparticles in the composition is no greater than about 200nm (such as less than about 200nm, for example no greater than about 100 nm). In some embodiments, there is provided a method of delivering a composition comprising nanoparticles comprising albumin and a macrolide (such as rapamycin) to a blood vessel, wherein the method comprises injecting (e.g. via a catheter with a needle) into the tissue surrounding the blood vessel wall an effective amount of a composition comprising nanoparticles comprising rapamycin and an albumin (such as human serum albumin), wherein the rapamycin in the nanoparticles is coated with the albumin, and wherein the average particle size of the nanoparticles in the composition is no greater than about 200nm (such as less than about 200nm, for example no greater than about 100 nm). In some embodiments, there is provided a method of delivering Nab-rapamycin to a blood vessel, wherein the method comprises injecting (e.g., via a catheter with a needle) an effective amount of Nab-rapamycin into the tissue surrounding the blood vessel wall. In some embodiments, the nanoparticle composition is injected at a disease site. In some embodiments, the nanoparticle composition is injected distal to the disease site (e.g., at least about any of: 1, 2, 3, 4, 5, 6,7, 8, 9, or 10cm from the disease site).

In some embodiments, the nanoparticle composition is injected into adventitial tissue. Adventitial tissue is perivascular tissue, such as tissue outside the outer elastic layer of an artery or tissue outside the media of a vein. The outer membrane has a high concentration of lipids. In some embodiments, the nanoparticle composition is injected into the vasa vasorum region of the adventitia. In some embodiments, the nanoparticle composition, after injection, may be dispersed from the injection site through the adventitia, circumferentially, longitudinally, and/or transmurally relative to the axis of the blood vessel into which the nanoparticle composition is injected (hereinafter referred to as "volume distribution"). In some embodiments, the drug (albumin-bound form or nanoparticle form) is distributed at a distance of at least about 1cm longitudinally (e.g., at least about any of: 2cm, 3cm, 4cm, 5cm, 6cm, 7cm or more) and/or at least 1cm radially (e.g., at least about any of: 2cm, 3cm, 4cm, 5cm, 6cm, 7cm or more) from the injection site over a period of no more than 60 minutes. In some embodiments, the drug concentration measured at all locations at least 2cm from the delivery site is at least 10% (e.g., at least about any of: 20%, 30%, 40%, or 50%) of the delivery site concentration-e.g., after a 60 minute period of time. In some embodiments, the drug (albumin bound or nanoparticle form) is transmurally distributed throughout the vascular endothelium and intimal, intermediate and muscle layers. While periadventitial administration of agents has been reported to allow for volume distribution of agents, it is believed that larger substances are not effectively distributed because volume distribution is achieved through the lymphatic microcirculatory system surrounding the blood vessels. The behaviour of the nanoparticle composition in the adventitial tissue is unknown. The present invention therefore differs in these respects from the methods reported.

Thus, in some embodiments, there is provided a method of delivering a composition comprising nanoparticles comprising albumin and a macrolide (such as rapamycin) to a blood vessel, wherein the method comprises injecting (e.g. via a catheter with a needle) into adventitial tissue of the blood vessel wall an effective amount of a composition comprising nanoparticles comprising a macrolide and albumin. In some embodiments, there is provided a method of delivering a composition comprising nanoparticles comprising albumin and a macrolide (such as rapamycin) to a blood vessel, wherein the method comprises injecting (e.g. via a catheter with a needle) into the adventitial tissue of the blood vessel an effective amount of a composition comprising nanoparticles comprising a macrolide and an albumin (such as human serum albumin), wherein the macrolide in the nanoparticles is coated with the albumin. In some embodiments, there is provided a method of delivering a composition comprising nanoparticles comprising albumin and a macrolide (such as rapamycin) to a blood vessel, wherein the method comprises injecting (e.g. via a catheter with a needle) into adventitial tissue of the blood vessel an effective amount of a composition comprising nanoparticles comprising a macrolide and an albumin (such as human serum albumin), wherein the average particle size of the nanoparticles in the composition is no greater than about 200nm (such as less than about 200nm, for example no greater than about 100 nm). In some embodiments, there is provided a method of delivering a composition comprising nanoparticles comprising albumin and a macrolide (such as rapamycin) to a blood vessel, wherein the method comprises injecting (e.g. via a catheter with a needle) into adventitial tissue of the blood vessel an effective amount of a composition comprising nanoparticles comprising rapamycin and albumin (such as human serum albumin), wherein the rapamycin in the nanoparticles is coated with albumin, and wherein the average particle size of the nanoparticles in the composition is no greater than about 200nm (such as less than about 200nm, for example no greater than about 100 nm). In some embodiments, there is provided a method of delivering Nab-rapamycin to a blood vessel, wherein the method comprises injecting (e.g., via a catheter with a needle) an effective amount of Nab-rapamycin into adventitial tissue of the blood vessel wall. In some embodiments, the nanoparticle composition is injected at or near the disease site (e.g., no more than about 2, 1, or 0.5cm from the disease site). In some embodiments, the nanoparticle composition is injected away from the disease site (e.g., at least about any of: 1, 2, 3, 4, 5, 6,7, 8, 9, or 10cm from the disease site). In some embodiments, the nanoparticle composition, upon injection, achieves a volume distribution.

In some embodiments the blood vessel is an artery, such as a coronary artery or a peripheral artery. In some embodiments, the artery is selected from the group consisting of a renal artery, a cerebral artery, a pulmonary artery, and a leg artery. In some embodiments, the blood vessel is an artery or vein above the knee. In some embodiments, the blood vessel is an artery or vein below the knee. In some embodiments, the blood vessel is a femoral artery. In some embodiments, the blood vessel is a balloon-injured artery.

In some embodiments, the blood vessel is an artery selected from any one of the following: the abdominal aorta, anterior tibial artery, aortic arch, arcus, axillary artery, brachial artery, carotid artery, abdominal artery, perifibular artery, common hepatic artery, common iliac artery, deep femoral artery, deep metacarpal artery arch, dorsal digital/phalangeal artery, dorsal metatarsal artery, external carotid artery, external iliac artery, facial artery, femoral artery, inferior mesenteric artery, internal iliac artery, intestinal artery, lateral below knee artery, lateral above knee artery, digital/metacarpal artery, peroneal artery, popliteal artery, posterior tibial artery, deep femoral artery, pulmonary artery, radial artery, renal artery, splenic artery, subclavian artery, superficial metacarpal artery arch, superior mesenteric artery, superior ulnar artery, and ulnar artery.

In some embodiments, the blood vessel is a vein. In some embodiments, the blood vessel is a vein selected from any one of the following: minor cephalic vein, axillary vein, basilic vein, brachial vein, cephalic vein, common iliac vein, dorsal digital/ungual vein, dorsal metatarsal vein, external iliac vein, facial vein, femoral vein, great saphenous vein, hepatic vein, inferior mesenteric vein, inferior vena cava, middle forearm vein, internal iliac vein, intestinal vein, jugular vein, lateral circumflex femoral vein, left inferior pulmonary vein, left superior pulmonary vein, digital/metacarpal vein, portal vein, posterior tibial vein, renal vein, posterior mandibular vein, saphenous vein, small saphenous vein, splenic vein, subclavian vein, superior mesenteric vein, and superior vena cava.

In some embodiments, the blood vessel is part of the coronary vasculature (including arterial and venous vasculature), cerebrovascular system, hepatic vasculature, peripheral vasculature, and other organ and tissue compartmentalized vasculature.

In some embodiments, there is provided a method of delivering a composition comprising nanoparticles comprising albumin and a macrolide (such as rapamycin or a derivative thereof, e.g. rapamycin) to a blood vessel, wherein the method comprises periadventitially injecting (e.g. via a catheter with a needle) (i.e. into periadventitial tissue) into the femoral artery an effective amount of a composition comprising nanoparticles comprising a macrolide and albumin. In some embodiments, there is provided a method of delivering a composition comprising nanoparticles comprising albumin and a macrolide (such as rapamycin or a derivative thereof, e.g. rapamycin) to a blood vessel, wherein the method comprises periadventitially injecting (e.g. via a catheter with a needle) into the femoral artery an effective amount of a composition comprising nanoparticles comprising a macrolide and an albumin (such as human serum albumin), wherein the macrolide in the nanoparticles is coated with the albumin. In some embodiments, there is provided a method of delivering a composition comprising nanoparticles comprising albumin and a macrolide (such as rapamycin or a derivative thereof, e.g. rapamycin) to a blood vessel, wherein the method comprises periadventitially injecting (e.g. via a catheter with a needle) into the femoral artery an effective amount of a composition comprising nanoparticles comprising a macrolide and an albumin (such as human serum albumin), wherein the average particle size of the nanoparticles in the composition is no greater than about 200nm (such as less than about 200nm, for example no greater than about 100 nm). In some embodiments, there is provided a method of delivering a composition comprising nanoparticles comprising albumin and a macrolide (such as rapamycin or a derivative thereof, e.g. rapamycin) to a blood vessel, wherein the method comprises periadventitially injecting (e.g. via a catheter with a needle) into the femoral artery an effective amount of a composition comprising nanoparticles comprising rapamycin and an albumin (such as human serum albumin), wherein the rapamycin in the nanoparticles is coated with the albumin, and wherein the average particle size of the nanoparticles in the composition is no greater than about 200nm (such as less than about 200nm, for example no greater than about 100 nm). In some embodiments, there is provided a method of delivering Nab-rapamycin to a blood vessel, wherein the method comprises periadventitially injecting (e.g., via a catheter with a needle) an effective amount of Nab-rapamycin to the femoral artery. In some embodiments, the nanoparticle composition is injected at or near the disease site (e.g., no more than about 2, 1, or 0.5cm from the disease site). In some embodiments, the nanoparticle composition is injected away from the disease site (e.g., at least about any of: 1, 2, 3, 4, 5, 6,7, 8, 9, or 10cm from the disease site).

The delivery methods described herein are effective to inhibit one or more aspects of vascular abnormalities, including, for example, negative remodeling, vascular fibrosis, restenosis, cell proliferation and cell migration in blood vessels, and wound healing. In some embodiments, the method is effective to promote positive remodeling of blood vessels.

The present application thus provides, in some embodiments, a method of inhibiting negative remodeling in a blood vessel in an individual in need thereof, comprising injecting into the blood vessel wall or tissue surrounding the blood vessel wall an effective amount of a composition comprising nanoparticles comprising a macrolide (such as rapamycin) and an albumin. In some embodiments, there is provided a method of inhibiting negative remodeling in a blood vessel in an individual in need thereof, comprising injecting into the blood vessel wall or tissue surrounding the blood vessel wall an effective amount of a composition comprising nanoparticles comprising a macrolide and an albumin (such as human serum albumin), wherein the macrolide in the nanoparticles is coated with the albumin. In some embodiments, there is provided a method of inhibiting negative remodeling in a blood vessel in an individual in need thereof, comprising injecting into the blood vessel wall or tissue surrounding the blood vessel wall an effective amount of a composition comprising nanoparticles comprising a macrolide and an albumin (such as human serum albumin), wherein the average particle size of the nanoparticles in the composition is no greater than about 200nm (such as less than about 200nm, for example no greater than about 100 nm). In some embodiments, there is provided a method of inhibiting negative remodeling in a blood vessel in an individual in need thereof, comprising injecting into the blood vessel wall or tissue surrounding the blood vessel wall an effective amount of a composition comprising nanoparticles comprising rapamycin and albumin (such as human serum albumin), wherein the rapamycin in the nanoparticles is coated with albumin, and wherein the average particle size of the nanoparticles in the composition is no greater than about 200nm (such as less than about 200nm, for example, no greater than about 100 nm). In some embodiments, there is provided a method of inhibiting negative remodeling in a blood vessel in an individual in need thereof, comprising injecting into the blood vessel wall or tissue surrounding the blood vessel wall an effective amount of Nab-rapamycin. In some embodiments, the nanoparticle composition is injected at or adjacent to the negative remodeling site (e.g., no more than about 2, 1, or 0.5cm from the negative remodeling site). In some embodiments, the nanoparticle composition is injected away from the negative remodeling site (e.g., at least about any of: 1, 2, 3, 4, 5, 6,7, 8, 9, or 10cm from the negative remodeling site). In some embodiments, the injection is through a catheter with a needle.

Negative remodeling involves the physiological or pathological response of the vessel to a stimulus, resulting in a reduction in vessel diameter and lumen diameter. Such stimulation may be provided by, for example, blood flow changes or angioplasty procedures. In some embodiments, injection of the nanoparticle composition results in an increase in the diameter of the blood vessel compared to the diameter of an uninjected blood vessel of about any of the following: 10%, 20%, 30%, 40%, 60%, 70%, 80%, 95% or more. Negative remodeling can be quantified, for example, by angiography as the percentage of diametric stenosis at the site of injury (or disease). Another method of determining the extent of remodeling involves measuring the area of the external elastic layer within the lesion using intravascular ultrasound (IVUS). IVUS is a technique that can image the external elastic layer as well as the lumen of the blood vessel. In some embodiments, the negative remodeling is associated with a vascular interventional procedure such as angioplasty, stenting, or atherectomy. The nanoparticle composition may thus be injected during or after the vascular interventional procedure.

In some embodiments, there is provided a method of promoting positive remodeling in a blood vessel in an individual in need thereof, comprising injecting into the blood vessel wall or tissue surrounding the blood vessel wall an effective amount of a composition comprising nanoparticles comprising a macrolide (such as rapamycin) and an albumin. In some embodiments, there is provided a method of promoting positive remodeling in a blood vessel in an individual in need thereof, comprising injecting into the blood vessel wall or tissue surrounding the blood vessel wall an effective amount of a composition comprising nanoparticles comprising a macrolide and an albumin (such as human serum albumin), wherein the macrolide in the nanoparticles is coated with the albumin. In some embodiments, there is provided a method of promoting positive remodeling in a blood vessel in an individual in need thereof, comprising injecting into the blood vessel wall or tissue surrounding the blood vessel wall an effective amount of a composition comprising nanoparticles comprising a macrolide and an albumin (such as human serum albumin), wherein the average particle size of the nanoparticles in the composition is no greater than about 200nm (such as less than about 200nm, for example no greater than about 100 nm). In some embodiments, there is provided a method of promoting positive remodeling in a blood vessel in an individual in need thereof, comprising injecting into the blood vessel wall or tissue surrounding the blood vessel wall an effective amount of a composition comprising nanoparticles comprising rapamycin and albumin (such as human serum albumin), wherein the rapamycin in the nanoparticles is coated with albumin, and wherein the average particle size of the nanoparticles in the composition is no greater than about 200nm (such as less than about 200nm, for example, no greater than about 100 nm). In some embodiments, there is provided a method of promoting positive remodeling in a blood vessel in an individual in need thereof, comprising injecting into the blood vessel wall or tissue surrounding the blood vessel wall an effective amount of Nab-rapamycin. In some embodiments, the nanoparticle composition is injected at or adjacent to the negative remodeling site (e.g., no more than about 2, 1, or 0.5cm from the negative remodeling site). In some embodiments, the nanoparticle composition is injected away from the negative remodeling site (e.g., at least about any of: 1, 2, 3, 4, 5, 6,7, 8, 9, or 10cm from the negative remodeling site). In some embodiments, the injection is through a catheter with a needle.

As used herein, positive remodeling refers to an increase in the diameter of a blood vessel compared to the diameter of a blood vessel without injection. In some embodiments, injection of the nanoparticle composition results in an increase in the diameter of the blood vessel compared to the diameter of a blood vessel without injection of about any of the following: 10%, 20%, 30%, 40%, 60%, 70%, 80%, 95% or more.

In some embodiments, there is provided a method of inhibiting vascular fibrosis (e.g., medial vascular fibrosis) in a blood vessel of an individual in need thereof, comprising injecting into the blood vessel wall or tissue surrounding the blood vessel wall an effective amount of a composition comprising nanoparticles comprising a macrolide (e.g., rapamycin) and an albumin. In some embodiments, there is provided a method of inhibiting vascular fibrosis (such as medial vascular fibrosis) in a blood vessel of an individual in need thereof, comprising injecting into the blood vessel wall or tissue surrounding the blood vessel wall an effective amount of a composition comprising nanoparticles comprising a macrolide and an albumin (such as human serum albumin), wherein the macrolide in the nanoparticles is coated with the albumin. In some embodiments, there is provided a method of inhibiting vascular fibrosis (such as medial vascular fibrosis) in a blood vessel of an individual in need thereof, comprising injecting into the blood vessel wall or tissue surrounding the blood vessel wall an effective amount of a composition comprising nanoparticles comprising a macrolide and an albumin (such as human serum albumin), wherein the average particle size of the nanoparticles in the composition is no greater than about 200nm (such as less than about 200nm, for example no greater than about 100 nm). In some embodiments, there is provided a method of inhibiting vascular fibrosis (such as medial vascular fibrosis) in a blood vessel of an individual in need thereof, comprising injecting into the blood vessel wall or tissue surrounding the blood vessel wall an effective amount of a composition comprising nanoparticles comprising rapamycin and albumin (such as human serum albumin), wherein the rapamycin in the nanoparticles is coated with the albumin, and wherein the average particle size of the nanoparticles in the composition is no greater than about 200nm (such as less than about 200nm, for example no greater than about 100 nm). In some embodiments, there is provided a method of inhibiting vascular fibrosis (e.g., medial vascular fibrosis) in a blood vessel of an individual in need thereof, comprising injecting into the blood vessel wall or tissue surrounding the blood vessel wall an effective amount of Nab-rapamycin. In some embodiments, the nanoparticle composition is injected at or adjacent to the site of vascular fibrosis (e.g., no more than about any of 2, 1, 0.5cm from the site of vascular fibrosis). In some embodiments, the nanoparticle composition is injected away from the site of vascular fibrosis (e.g., at least about any of: 2, 3, 4, 5, 6,7, 8, 9, or 10cm from the site of vascular fibrosis). In some embodiments, the injection is through a catheter with a needle.

As used herein, vascular fibrosis refers to the formation of a wide range of fibrous (connective) tissues in the blood vessel, including, for example, medial fibrosis or adventitial fibrosis. Vascular fibrosis is often associated with massive extracellular matrix deposition and myofibroblast and fibroblast proliferation. Thus, the methods described herein, in some embodiments, inhibit fibrous tissue formation in a blood vessel, e.g., inhibit any of the following compared to an uninjected blood vessel: 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80% or 90% fibrous tissue formation. In some embodiments, the method inhibits extracellular matrix deposition in a blood vessel, e.g., inhibits any of the following compared to an uninjected blood vessel: 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80% or 90% extracellular matrix deposition. In some embodiments, the method inhibits myofibroblast proliferation in a blood vessel, e.g., inhibits any of the following compared to an uninjected blood vessel: 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80% or 90% myofibroblast proliferation. In some embodiments, the method inhibits proliferation of fibroblasts in a blood vessel, e.g., inhibits any of the following compared to a non-injected blood vessel: 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80% or 90% fibroblast proliferation. In some embodiments, the vascular fibrosis is associated with a vascular interventional procedure such as angioplasty, stenting, or atherectomy. The nanoparticle composition may thus be injected during or after the vascular interventional procedure.

Thus, in some embodiments, the methods described herein inhibit luminal stenosis, e.g., inhibit any of the following compared to an uninjected vessel: 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80% or 90% luminal stenosis. In some embodiments, the luminal stenosis is associated with a vascular interventional procedure such as angioplasty, stenting, or atherectomy. The nanoparticle composition may thus be injected during or after the vascular interventional procedure.

In some embodiments, there is provided a method of treating restenosis in a blood vessel in an individual in need thereof, comprising injecting into the blood vessel wall or tissue surrounding the blood vessel wall an effective amount of a composition comprising nanoparticles comprising a macrolide (such as rapamycin) and an albumin. In some embodiments, there is provided a method of treating restenosis in a blood vessel in an individual in need thereof, comprising injecting into the blood vessel wall or tissue surrounding the blood vessel wall an effective amount of a composition comprising nanoparticles comprising a macrolide and an albumin (such as human serum albumin), wherein the macrolide in the nanoparticles is coated with the albumin. In some embodiments, there is provided a method of treating restenosis in a blood vessel in an individual in need thereof, comprising injecting into the blood vessel wall or tissue surrounding the blood vessel wall an effective amount of a composition comprising nanoparticles comprising a macrolide and an albumin (such as human serum albumin), wherein the average particle size of the nanoparticles in the composition is no greater than about 200nm (such as less than about 200nm, for example no greater than about 100 nm). In some embodiments, there is provided a method of treating restenosis in a blood vessel in an individual in need thereof, comprising injecting into the blood vessel wall or tissue surrounding the blood vessel wall an effective amount of a composition comprising nanoparticles comprising rapamycin and albumin (such as human serum albumin), wherein the rapamycin in the nanoparticles is coated with albumin, and wherein the average particle size of the nanoparticles in the composition is no greater than about 200nm (such as less than about 200nm, for example, no greater than about 100 nm). In some embodiments, there is provided a method of treating restenosis in a blood vessel in an individual in need thereof, comprising injecting into the blood vessel wall or tissue surrounding the blood vessel wall an effective amount of Nab-rapamycin. In some embodiments, the nanoparticle composition is injected at or near the disease site (e.g., no more than about 2, 1, or 0.5cm from the disease site). In some embodiments, the nanoparticle composition is injected away from the disease site (e.g., at least about any of: 1, 2, 3, 4, 5, 6,7, 8, 9, or 10cm from the disease site). In some embodiments, the injection is through a catheter with a needle.

In some embodiments, there is provided a method of reducing adventitial leukocytes in a blood vessel in an individual in need thereof, comprising injecting into the blood vessel wall or tissue surrounding the blood vessel wall an effective amount of a composition comprising nanoparticles comprising a macrolide (such as rapamycin) and an albumin. In some embodiments, there is provided a method of reducing adventitial leukocytes in a blood vessel in an individual in need thereof, comprising injecting into the blood vessel wall or tissue surrounding the blood vessel wall an effective amount of a composition comprising nanoparticles comprising a macrolide and an albumin (such as human serum albumin), wherein the macrolide in the nanoparticles is coated with the albumin. In some embodiments, there is provided a method of reducing adventitial leukocytes in a blood vessel in an individual in need thereof, comprising injecting into the blood vessel wall or tissue surrounding the blood vessel wall an effective amount of a composition comprising nanoparticles comprising a macrolide and an albumin (such as human serum albumin), wherein the average particle size of the nanoparticles in the composition is no greater than about 200nm (such as less than about 200nm, for example no greater than about 100 nm). In some embodiments, there is provided a method of reducing adventitial leukocytes in a blood vessel in an individual in need thereof, comprising injecting into the blood vessel wall or tissue surrounding the blood vessel wall an effective amount of a composition comprising nanoparticles comprising rapamycin and albumin (such as human serum albumin), wherein the rapamycin in the nanoparticles is coated with albumin, and wherein the average particle size of the nanoparticles in the composition is no greater than about 200nm (such as less than about 200nm, for example, no greater than about 100 nm). In some embodiments, there is provided a method of reducing adventitial leukocytes in a blood vessel in an individual in need thereof, comprising injecting into the blood vessel wall or tissue surrounding the blood vessel wall an effective amount of Nab-rapamycin. In some embodiments, the nanoparticle composition is injected into adventitial tissue.

In some embodiments, there is provided a method of reducing adventitial vessels in a blood vessel of an individual in need thereof, comprising injecting into the blood vessel wall or tissue surrounding the blood vessel wall an effective amount of a composition comprising nanoparticles comprising a macrolide (such as rapamycin) and an albumin. In some embodiments, there is provided a method of reducing adventitial vessels of a blood vessel in an individual in need thereof, comprising injecting into the blood vessel wall or tissue surrounding the blood vessel wall an effective amount of a composition comprising nanoparticles comprising a macrolide and an albumin (such as human serum albumin), wherein the macrolide in the nanoparticles is coated with the albumin. In some embodiments, there is provided a method of reducing adventitial vessels in a blood vessel of an individual in need thereof, comprising injecting into the blood vessel wall or tissue surrounding the blood vessel wall an effective amount of a composition comprising nanoparticles comprising a macrolide and an albumin (such as human serum albumin), wherein the average particle size of the nanoparticles in the composition is no greater than about 200nm (such as less than about 200nm, for example no greater than about 100 nm). In some embodiments, there is provided a method of reducing adventitial vessels of a blood vessel in an individual in need thereof, comprising injecting into the blood vessel wall or tissue surrounding the blood vessel wall an effective amount of a composition comprising nanoparticles comprising rapamycin and albumin (such as human serum albumin), wherein the rapamycin in the nanoparticles is coated with albumin, and wherein the average particle size of the nanoparticles in the composition is no greater than about 200nm (such as less than about 200nm, for example no greater than about 100 nm). In some embodiments, there is provided a method of reducing adventitial vessels in a blood vessel in an individual in need thereof, comprising injecting into the blood vessel wall or tissue surrounding the blood vessel wall an effective amount of Nab-rapamycin. In some embodiments, the nanoparticle composition is injected into adventitial tissue.

In some embodiments, the individual is a human. In some embodiments, the subject is at least about any of: 35. 40, 45, 50, 55, 60, 65, 70, 75, 80, or 85 years old. In some embodiments, the individual has asian descent. In some embodiments, the subject is a male. In some embodiments, the subject is a female. In some embodiments, the subject has a disease described below.

The methods described herein can be used to treat a variety of diseases. These include, for example, angina, aortic stenosis, arteriosclerosis obliterans, carotid stenosis, cerebrovascular artery disease, cerebrovascular occlusion, coronary artery disease, dilated cardiomyopathy, ischemic cardiomyopathy, intermittent claudication, peripheral arterial stenosis, renal artery disease, restenosis, small vessel disease, stenosis, aortic valve stenosis, arteriosclerotic transluminal sclerosis, proliferative arteriosclerosis, mitral valve stenosis, pulmonary valve stenosis, tricuspid valve stenosis, deep vein thrombosis, peripheral venous disease, and thrombophlebitis. The methods described herein may include treating any one or more of these diseases.

In some embodiments, the disease is selected from angina, myocardial infarction, congestive heart failure, cardiac arrhythmia, peripheral arterial disease, claudication, or chronic limb ischemia. Thus, for example, in some embodiments, there is provided a method of treating angina (or myocardial infarction or congestive heart failure or arrhythmia or peripheral artery disease or claudication or chronic limb ischemia) in an individual in need thereof, comprising injecting into the blood vessel wall or tissue surrounding the blood vessel wall an effective amount of a composition comprising nanoparticles comprising a macrolide (such as rapamycin) and an albumin. In some embodiments, there is provided a method of treating angina (or myocardial infarction or congestive heart failure or arrhythmia or peripheral arterial disease or claudication or chronic limb ischemia) in an individual in need thereof, comprising injecting into the blood vessel wall or tissue surrounding the blood vessel wall an effective amount of a composition comprising nanoparticles comprising a macrolide and an albumin (such as human serum albumin), wherein the macrolide in the nanoparticles is coated with the albumin. In some embodiments, there is provided a method of treating angina (or myocardial infarction or congestive heart failure or arrhythmia or peripheral arterial disease or claudication or chronic limb ischemia) in an individual in need thereof, comprising injecting into the blood vessel wall or tissue surrounding the blood vessel wall an effective amount of a composition comprising nanoparticles comprising a macrolide and an albumin (such as human serum albumin), wherein the average particle size of the nanoparticles in the composition is no greater than about 200nm (such as less than about 200nm, for example no greater than about 100 nm). In some embodiments, there is provided a method of treating angina (or myocardial infarction or congestive heart failure or arrhythmia or peripheral arterial disease or claudication or chronic limb ischemia) in an individual in need thereof, comprising injecting into the blood vessel wall or tissue surrounding the blood vessel wall an effective amount of a composition comprising nanoparticles comprising rapamycin and albumin (such as human serum albumin), wherein the rapamycin in the nanoparticles is coated with albumin, and wherein the average particle size of the nanoparticles in the composition is no greater than about 200nm (such as less than about 200nm, for example, no greater than about 100 nm). In some embodiments, there is provided a method of treating angina (or myocardial infarction or congestive heart failure or arrhythmia or peripheral arterial disease or claudication or chronic limb ischemia) in an individual in need thereof, comprising injecting into the blood vessel wall or tissue surrounding the blood vessel wall an effective amount of Nab-rapamycin. In some embodiments, the nanoparticle composition is injected at or near the disease site (e.g., no more than about 2, 1, or 0.5cm from the disease site). In some embodiments, the nanoparticle composition is injected away from the disease site (e.g., at least about any of: 1, 2, 3, 4, 5, 6,7, 8, 9, or 10cm from the disease site). In some embodiments, the injection is through a catheter with a needle. In some embodiments, the nanoparticle composition is injected during or after a vascular interventional procedure such as angioplasty, stenting, or atherectomy.

In some embodiments, the methods described herein comprise injecting the nanoparticle composition distal to the disease site. In some embodiments, the nanoparticle composition is injected proximal to the disease site. The delivery site may be in the same vessel as the disease treatment region, longitudinally spaced from the region, or may be in a different vessel. In some embodiments, the nanoparticle composition is injected at or adjacent to the disease site (e.g., no more than about any of 2, 1, or 0.5cm from (e.g., longitudinally from) the disease site). In some embodiments, the nanoparticle composition is injected away from the disease site (e.g., about any of 1, 2, 3, 4, 5, 6,7, 8, 9, or 10cm from (e.g., longitudinally from) the disease site). In some embodiments, the disease treatment area may have previously been implanted with a stent, with the delivery site spaced from the stent-longitudinally distal to the stent in the same coronary artery or distal to the stent in another coronary artery or vein.

In some embodiments, the methods described herein comprise injecting the nanoparticle composition with a needle (e.g., a deployable needle). The needle can be positioned such that the nanoparticle composition is delivered to a desired site. Thus, in some embodiments, the method comprises positioning a needle, penetrating a blood vessel wall, and delivering an effective amount of a nanoparticle composition into the blood vessel wall or tissue surrounding the blood vessel wall. For example, in some embodiments, the needle hole is located outside the extravascular elastic layer such that the nanoparticle composition is delivered to adventitial tissue surrounding the blood vessel. In some embodiments, the needle hole is positioned at a distance no greater than about 0.1mm, about 0.2mm, 0.5mm, 0.8mm, 1cm, 2cm, 3cm, 4cm, 5cm, 6cm, 7cm, or 8cm outside of the extravascular elastic layer.

In some embodiments, the holes are positioned at a distance from the inner wall of the blood vessel that is at least about 10% (including, e.g., at least about 20%, 30%, 40%, 60%, 70%, 80%, 90%) of the average lumen diameter of the blood vessel at the injection site. In some embodiments, the holes are positioned at a distance from the inner wall of the blood vessel that is about 10% to about 75% (including, e.g., about 20% to about 60%, about 30% to about 50%) of the average lumen diameter of the blood vessel at the injection site.

The determination of the position of the pinhole can be achieved in a number of ways. For example, after initial positioning of the needle is achieved, a bolus of contrast media or other visual medium may be injected through the needle. By observing the medium distribution, e.g. by fluoroscopy, the pore location can be estimated. In some embodiments, various sensors may be attached or otherwise coupled to the needle, typically near the delivery aperture, to detect the position of the needle. Exemplary sensors include temperature sensors, pH sensors, electrical impedance sensors, and the like. The back pressure against the injected suspension can also be measured to determine the needle position. Injection into the vessel wall will generally create a greater backpressure than injection into the adventitial space. The insertion force of the needle may also be monitored, for example, by providing a deflectometer on a portion of the needle.

Administration and method of administering nanoparticle compositions

The dose of macrolide nanoparticle composition injected into an individual (e.g., human) may vary depending on the type of injection vessel, the purpose of the method, and the type of disease being treated. In some embodiments, the injected amount of macrolide nanoparticle composition is sufficient to inhibit negative remodeling by more than about any of: 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 64%, 65%, 70%, 75%, 80%, 85% or 90%. Inhibition of negative remodeling can be assessed, for example, by measuring the vessel or lumen diameter of the blood vessel. In some embodiments, the injected amount of macrolide nanoparticle composition is sufficient to promote positive remodeling greater than about any of: 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 64%, 65%, 70%, 75%, 80%, 85% or 90%.

In some embodiments, the macrolide nanoparticle composition is injected in an amount sufficient to inhibit vascular fibrosis greater than about any of: 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 64%, 65%, 70%, 75%, 80%, 85% or 90%. In some embodiments, the vascular fibrosis is medial fibrosis. In some embodiments, the vascular fibrosis is adventitial fibrosis. Inhibition of vascular fibrosis can be assessed, for example, by assessing the amount of extracellular matrix deposition and/or myofibroblast and fibroblast proliferation. In some embodiments, vascular fibrosis is assessed by histopathological analysis, for example by staining with H & E or trichrome.

In some embodiments, the macrolide nanoparticle composition is injected in an amount sufficient to decrease the proliferation index by more than about any of the following: 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 64%, 65%, 70%, 75%, 80%, 85% or 90%. In some embodiments, the macrolide nanoparticle composition is injected in an amount sufficient to reduce luminal stenosis by more than about any of: 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 64%, 65%, 70%, 75%, 80%, 85% or 90%. In some embodiments, the macrolide nanoparticle composition is injected in an amount sufficient to reduce adventitial leukocytes by greater than about any of: 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 64%, 65%, 70%, 75%, 80%, 85% or 90%. In some embodiments, the macrolide nanoparticle composition is injected in an amount sufficient to reduce adventitial vessels by more than about any of: 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 64%, 65%, 70%, 75%, 80%, 85% or 90%.

In some embodiments, the amount of macrolide (e.g., rapamycin) in the composition is below a level that induces a toxicological effect (i.e., an effect above a clinically acceptable toxicity level) or at a level where potential side effects can be controlled or tolerated when the composition is injected into an individual.

In some embodiments, the unit injected amount of macrolide (e.g., rapamycin or a derivative thereof, e.g., rapamycin) is within any of the following ranges: about 0.001 to about 100mg, including, for example, about 0.001 to about 0.005mg, about 0.005 to about 0.025mg, about 0.025 to about 0.1mg, about 0.1 to about 0.5mg, about 0.5 to about 1mg, about 1 to about 2mg, about 2 to about 3mg, about 3 to about 4mg, about 4 to about 5mg, about 5 to about 6mg, about 6 to about 7mg, about 7 to about 8mg, about 8 to about 9mg, about 9 to about 10mg, about 10 to about 15mg, about 15 to about 20mg, about 20 to about 25mg, about 20 to about 50mg, about 25 to about 50mg, about 50 to about 75mg, or about 50 to about 100 mg. In some embodiments, the unit injected amount of macrolide (e.g., rapamycin) is in the following range: from about 0.001 to about 100mg, such as from about 0.005 to about 80mg, from about 0.05 to about 50mg, from about 0.1 to about 10mg, from about 0.1 to about 5mg, from about 0.5 to about 5mg, from about 0.05 to about 5mg, or from about 0.5 to about 2 mg.

In some embodiments, the macrolide (e.g., rapamycin) concentration in the nanoparticle composition is diluted (about 0.1mg/ml) or concentrated (about 100mg/ml), including, for example, any of the following: about 0.1 to about 50mg/ml, about 0.1 to about 20mg/ml, about 1 to about 10mg/ml, about 2mg/ml to about 8mg/ml, about 4 to about 6mg/ml, or about 5 mg/ml. In some embodiments, the concentration of macrolide (e.g., rapamycin) is at least about any of the following: 0.5mg/ml, 1.3mg/ml, 1.5mg/ml, 2mg/ml, 3mg/ml, 4mg/ml, 5mg/ml, 6mg/ml, 7mg/ml, 8mg/ml, 9mg/ml, 10mg/ml, 15mg/ml, 20mg/ml, 25mg/ml, 30mg/ml, 40mg/ml or 50 mg/ml. In some embodiments, the concentration of macrolide (e.g., rapamycin) is no greater than about any of the following: 100mg/ml, 90mg/ml, 80mg/ml, 70mg/ml, 60mg/ml, 50mg/ml, 40mg/ml, 30mg/ml, 20mg/ml, 10mg/ml or 5 mg/ml.

The unit injection volume of the nanoparticle composition may vary depending on the type of vessel injected, the purpose of the method, and the type of disease being treated. In some embodiments, the unit injection volume is from about 0.01 to about 50ml, including, for example, from about 0.01 to about 0.05ml, from about 0.05 to about 0.1ml, from about 0.1 to about 0.5ml, from about 0.5 to about 1ml, from about 1 to about 2ml, from about 2 to about 3ml, from about 3 to about 5ml, from about 5 to about 10ml, from about 10 to about 20ml, from about 20 to about 30ml, from about 30 to about 40ml, from about 40 to about 50 ml. In some embodiments, the unit injection volume is about 0.05 to about 2ml, about 0.1 to 1ml, about 0.25 to about 0.5ml, or about 0.5 to about 1ml, or about 1 to about 5 ml.

Exemplary dosing frequencies for administering the nanoparticle composition include, but are not limited to, about once every 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 11, or 12 months. In some embodiments, the interval between doses is greater than about any of the following: 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 8 months, or 12 months. In some embodiments, the administration is administered every 3, 6, 9, 12, 15, 18, 21, or 24 months. In some embodiments, the administration is administered at most every 3, 6, 9, 12, 15, 18, 21, or 24 months. In some embodiments, the administration is given at least every 3, 6, 9, 12, 15, 18, 21, or 24 months. In some embodiments, the nanoparticle composition is injected only once.

The nanoparticle composition may be injected during vascular interventional procedures. In some embodiments, the nanoparticle composition is injected once during the vascular interventional procedure. In some embodiments, the nanoparticle composition is injected after vascular interventional surgery. Exemplary vascular interventional procedures include, but are not limited to, angioplasty, stent implantation, and atherectomy.

Nanoparticle compositions

The nanoparticle compositions described herein include nanoparticles that include (in various embodiments consist essentially of or consist of) a macrolide (such as rapamycin) and an albumin (such as human serum albumin). Nanoparticles of poorly water soluble drugs (such as macrolides) have been disclosed, for example, in U.S. Pat. nos. 5,916,596; 6,506,405, respectively; 6,749,868, 6,537,579, 7,820,788, and U.S. patent publication nos. 2006/0263434, and 2007/0082838; PCT patent application WO08/137148, the entire contents of which are incorporated by reference.

In some embodiments, the composition comprises nanoparticles having an average (mean) diameter of no greater than about 1000 nanometers (nm), such as no greater than about any of the following: 900. 800, 700, 600, 500, 400, 300, 200 and 100 nm. In some embodiments, the nanoparticles have an average diameter of no greater than about 200 nm. In some embodiments, the nanoparticles have an average diameter of no greater than about 150 nm. In some embodiments, the nanoparticles have an average diameter of no greater than about 100 nm. In some embodiments, the nanoparticles have an average diameter of about 20 to about 400 nm. In some embodiments, the nanoparticles have an average diameter of about 40 to about 200 nm. In some embodiments, the nanoparticles have an average diameter of about 50 to about 100 nm. In some embodiments, the nanoparticles are not less than about 50 nm. In some embodiments, the nanoparticles may be sterile filtered.

In some embodiments, the average diameter of the nanoparticles in the compositions described herein is no greater than about 200nm, including, for example, no greater than any of about 190, 180, 170, 160, 150, 140, 130, 120, 110, 100, 90, 80, 70, or 60 nm. In some embodiments, at least about 50% (e.g., at least about any of 60%, 70%, 80%, 90%, 95%, or 99%) of the nanoparticles in the composition have a diameter no greater than about 200nm, including, for example, no greater than about any of 190, 180, 170, 160, 150, 140, 130, 120, 110, 100, 90, 80, 70, or 60 nm.

In some embodiments, the nanoparticles in the compositions described herein have an average diameter of no less than about 50nm, including, for example, no less than about any of 50, 60, 70, 80, 90, or 100 nm. In some embodiments, at least about 50% (e.g., at least about any of 60%, 70%, 80%, 90%, 95%, or 99%) of the nanoparticles in the composition have a diameter of no less than about 50nm, including, for example, no less than about any of 50, 60, 70, 80, 90, or 100 nm.

In some embodiments, at least about 50% (e.g., at least any of 60%, 70%, 80%, 90%, 95%, or 99%) of the nanoparticles in the composition fall within the following ranges: about 20 to about 400nm, including, for example, any of about 20 to about 200nm, about 40 to about 200nm, about 30 to about 180nm, and about 40 to about 150, about 50 to about 120, and about 60 to about 100 nm.

In some embodiments, the albumin has a thiol group that can form a disulfide bond. In some embodiments, at least about 5% (including, e.g., at least any of about 10%, 15%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 80%, or 90%) of the albumin in the nanoparticle portion of the composition is cross-linked (e.g., cross-linked by one or more disulfide bonds).

In some embodiments, the nanoparticle comprises a macrolide (such as rapamycin) coated with albumin (e.g., human serum albumin). In some embodiments, the composition comprises nanoparticles and macrolide in non-nanoparticle form (e.g., in the form of a rapamycin solution or in the form of a soluble albumin/nanoparticle complex), wherein at least about 50%, 60%, 70%, 80%, 90%, 95%, or 99% of the macrolide in any of the compositions are in nanoparticle form. In some embodiments, the macrolide in the nanoparticle constitutes greater than about any of 50%, 60%, 70%, 80%, 90%, 95%, or 99% by weight of the nanoparticle. In some embodiments, the nanoparticle has a non-polymeric matrix. In some embodiments, the nanoparticle comprises a macrolide core that is substantially free of polymer (e.g., polymeric matrix).

In some embodiments, the composition comprises albumin in the nanoparticle and non-nanoparticle portions of the composition, wherein at least about any one of 50%, 60%, 70%, 80%, 90%, 95%, or 99% of the albumin in the composition is in the non-nanoparticle portion of the composition.

In some embodiments, the weight ratio of albumin (such as human serum albumin) to macrolide in the 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 serum albumin) to macrolide in the composition falls within any one of the following ranges: 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 macrolide in the nanoparticle portion of the composition is about any of the following: 1:2, 1:3, 1:4, 1:5, 1:10, 1:15, or less. In some embodiments, the weight ratio of albumin (such as human serum albumin) to macrolide in the composition is any one 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, or about 1: 1.

In some embodiments, the nanoparticle composition includes one or more of the features described above.

The nanoparticles described herein can be present in a dry formulation (e.g., a lyophilized composition) or suspended in a biocompatible medium. Suitable biocompatible media include, but are not limited to, water, aqueous buffered media, saline, buffered saline, optionally amino acid buffered solutions, optionally protein buffered solutions, optionally sugar buffered solutions, optionally vitamin buffered solutions, optionally synthetic polymer buffered solutions, lipid containing emulsions, and the like.

In some embodiments, the pharmaceutically acceptable carrier comprises human serum albumin. Human Serum Albumin (HSA) is M_r65K, consisting of 585 amino acids. HAS is the most abundant protein in plasma, occupying 70-80% of the human plasma oncotic pressure. The amino acid sequence of HAS comprises a total of 17 disulfide bridges, 1 free thiol group (Cys34) and 1 tryptophan (Trp 214). Intravenous use of HSA solutions has been proposed for the prevention and treatment of hypovolemic shock (see, e.g., tulis, JAMA, 237, 355-360, 460-463, (1977)) and house et al, Surgery, gynecologane and obstercs, 150, 811-816(1980)), and in the treatment of neonatal hyperbilirubinemia in conjunction with exchange transfusion (see, e.g., Finlayson, Seminars in Thrombosis and hemostatis, 6, 85-120, (1980)). Other albumins are contemplated, such as bovine serum albumin. The use of such non-human albumin may be appropriate, for example, where the compositions are to be used in non-human mammals, such as veterinarians (including pets and agricultural contexts).

Human Serum Albumin (HSA) has multiple hydrophobic binding sites (8 in total, for fatty acids, HSA endogenous ligands) and binds to multiple macrolides, particularly neutral and negatively charged hydrophobic compounds (Goodmanet al, thermal Basis of therapeutics, 9)^thed, McGraw-Hill New York (1996)). Two high affinity binding sites in the subdomains IIA and IIIA of HAS been proposed, which are highly elongated hydrophobic pockets (pockets) with charged lysine and arginine residues near the surface, serving as attachment points for polar ligand moieties (see, e.g., Fehske et al, biochem. pharmacol., 30, 687-92(198a), Vorum, dan. med. bull., 46, 379-99(1999), Kragh-Hansen, dan. med. buzz., 1441, 131-40(1990), Curry et al, nat. struct. biol., 5, 827-35(1998), Sugio et al, protein. eng., 12, 439-46(1999), heal, Natur et al, for examplee, 358, 209-15(199b) and Carter et al, adv. protein. chem., 45, 153-. Rapamycin and propofol (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. biophysis.acta, 1478(a), 61-8(2000), Altmayer et al, arzneimitelforkung, 45, 1053-6(1995) and Garridoet et al, rev.e.estestrol.reanim., 41, 308-12 (1994)). In addition, docetaxel (docetaxel) has been shown to bind human plasma proteins (see, e.g., Urien et al, invest. New Drugs, 14(b), 147-51 (1996)).

The albumin (such as human serum albumin) in the composition typically acts as a carrier for the macrolide, i.e. the albumin in the composition makes the macrolide more readily suspendable in an aqueous medium or helps to remain suspended, as compared to a composition that does not comprise albumin. This may avoid the use of toxic solvents (or surfactants) for solubilizing the macrolide, and may reduce one or more side effects of administration of the macrolide to an individual (e.g., a human). Thus, in some embodiments, the compositions described herein are substantially free (e.g., free) of surfactants, such as Cremophor (including Cremophor)(BASF)). In some embodiments, the nanoparticle composition is substantially free (e.g., free) of surfactant. When the nanoparticle composition is injected into a subject, the composition is "substantially free of cremophor" or "substantially free of surfactant" if an insufficient amount of cremophor or surfactant in the composition produces one or more side effects in the subject. In some embodiments, the nanoparticle composition comprises less than about any of 20%, 15%, 10%, 7.5%, 5%, 2.5%, or 1% of an organic solvent or surfactant.

The amount of albumin in the compositions described herein will vary depending on the other components in the composition. In some embodiments, the composition comprises albumin in an amount sufficient to stabilize the macrolide in an aqueous suspension, e.g., in the form of a stable colloidal suspension (such as a stable nanoparticle suspension). In some embodiments, the amount of albumin is such that the rate of sedimentation of the macrolide in the aqueous medium is reduced. For particle-containing compositions, the amount of albumin also depends on the size and density of the macrolide nanoparticles.

The macrolide is "stabilized" in aqueous suspension if it remains suspended in the aqueous medium for a prolonged period of time (e.g., no visible precipitation or sedimentation), e.g., for a prolonged period of time of at least about any of the following: 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. Suspensions are generally, but not necessarily, suitable for administration to an individual (e.g., a human). The stability of the suspension is usually, but not necessarily, evaluated at storage temperature, such as room temperature (e.g. 20-25 ℃) or refrigerated storage conditions (e.g. 4 ℃). For example, the suspension is stable at storage temperatures if it does not exhibit flocculation or particle agglomeration visible to the naked eye or when viewed under a 1000-fold optical microscope about 15 minutes after suspension preparation. Stability may also be evaluated under accelerated test conditions, such as at temperatures above about 40 ℃.

In some embodiments, albumin is present in an amount sufficient to stabilize a particular concentration of macrolide in an aqueous suspension. For example, the macrolide concentration in the composition is from about 0.1 to about 100mg/ml, including for example any of the following: about 0.1 to about 50mg/ml, about 0.1 to about 20mg/ml, about 1 to about 10mg/ml, about 2mg/ml to about 8mg/ml, about 4 to about 6mg/ml, about 5 mg/ml. In some embodiments, the macrolide concentration is at least about any of the following: 1.3mg/ml, 1.5mg/ml, 2mg/ml, 3mg/ml, 4mg/ml, 5mg/ml, 6mg/ml, 7mg/ml, 8mg/ml, 9mg/ml, 10mg/ml, 15mg/ml, 20mg/ml, 25mg/ml, 30mg/ml, 40mg/ml, and 50 mg/ml. In some embodiments, the albumin is present in an amount that avoids the use of a surfactant (e.g., cremophor), such that the composition is free or substantially free of a surfactant (e.g., cremophor).

In some embodiments, the composition in liquid form 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)) albumin. In some embodiments, the composition in liquid form comprises from about 0.5% to about 5% (w/v) albumin.

In some embodiments, the weight ratio of albumin, e.g., albumin, to macrolide in the nanoparticle composition is such that a sufficient amount of macrolide binds to or is transported by the cell. Although the weight ratio of albumin to macrolide must be optimized for different albumin and macrolide combinations, typically the weight ratio (w/w) of albumin, e.g., albumin to macrolide is from about 0.01:1 to about 100:1, from about 0.02:1 to about 50:1, from about 0.05:1 to about 20:1, from about 0.1:1 to about 20:1, from about 1:1 to about 18:1, from about 2:1 to about 15:1, from about 3:1 to about 12:1, from about 4:1 to about 10:1, from about 5:1 to about 9:1, or about 9: 1. In some embodiments, the weight ratio of albumin to macrolide is about any of the following: 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 serum albumin) to macrolide in the composition is any one 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, or about 1: 1.

In some embodiments, the albumin enables the composition to be injected into an individual (e.g., a human) without significant side effects. In some embodiments, the amount of albumin (e.g., human serum albumin) is effective to reduce one or more side effects of macrolide administration to a human. The term "reducing one or more side effects of administration of a macrolide" means reducing, alleviating, eliminating or avoiding one or more of the unwanted effects caused by the macrolide as well as the side effects caused by the delivery vehicle used to deliver the macrolide (e.g., a solvent that renders the macrolide suitable for injection). Such side effects include, for example, myelosuppression, neurotoxicity, allergy, inflammation, venous irritation, phlebitis, pain, skin irritation, peripheral neuropathy, neutropenic fever (neutropenic lever), anaphylaxis, venous thrombosis, extravasation, and combinations thereof. However, these side effects are merely exemplary, and other side effects or combinations of side effects associated with macrolides may be reduced.

In some embodiments, the nanoparticle composition comprises Nab-rapamycin (Celgene Corp.). In some embodiments, the nanoparticle composition is Nab-rapamycin. Nab-rapamycin is a rapamycin formulation stabilized by human albumin USP, which can be dispersed in a physiological solution that can be directly injected. Nab-rapamycin forms a stable colloidal suspension of rapamycin when dispersed in a suitable aqueous medium such as 0.9% sodium chloride injection or 5% dextrose injection. The average particle size of the nanoparticles in the colloidal suspension was about 90 nm. Since HSA is freely soluble in water, Nab-rapamycin can be reconstituted at a wide range of concentrations from dilute (0.1mg/ml rapamycin) to concentrated (20mg/ml rapamycin), including for example, from about 2mg/ml to about 8mg/ml or about 5 mg/ml.

Methods of preparing nanoparticle compositions are known in the art. For example, nanoparticles comprising a macrolide (such as rapamycin) and an albumin (such as human serum albumin) can be prepared under conditions of high shear (e.g., sonication, high pressure homogenization, or similar forms). Such methods are disclosed, for example, in U.S. Pat. nos. 5,916,596; 6,506,405, respectively; 6,749,868, 6,537,579 and 7,820,788, and U.S. patent publication nos. 2007/0082838, 2006/0263434 and PCT application WO 08/137148.

Briefly, a macrolide (such as rapamycin) is dissolved in an organic solvent and the solution can be added to an albumin solution. The mixture was homogenized under high pressure. The organic solvent can then be removed by evaporation. The obtained dispersion may 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 may 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).

Other Components in the nanoparticle compositions

The negatively charged components include, but are not limited to, bile salts of bile acids consisting of glycocholic acid, cholic acid, chenodeoxycholic acid, taurocholic acid, glycochenodeoxycholic acid, taurochenodeoxycholic acid, lithocholic acid, deoxycholic acid, dehydrocholic acid, and others, phospholipids, including lecithin (egg yolk) type phospholipids, including phosphatidylcholines, palmitoyl oleoyl phosphatidylcholine, palmitoyl linoleoyl phosphatidylcholine, stearoyl oleoyl phosphatidylcholine, stearoyl arachidoyl phosphatidylcholine, and dipalmitoyl phosphatidylcholine.

In some embodiments, the composition is suitable for administration to a human. In some embodiments, the compositions are suitable for administration to mammals, such as, in the case of veterinarians, pets and agricultural animals. There are a variety of suitable nanoparticle composition formulations (see, e.g., U.S. Pat. nos. 5,916,596, 6,096,331, and 7,820,788). The following formulations and methods are exemplary only, and are in no way limiting. Formulations suitable for oral administration may consist of: (a) a liquid solution, such as an effective amount of the compound, dissolved in a diluent such as water, saline, or orange juice; (b) capsules, sachets or tablets each containing a predetermined amount of active ingredient as a solid or particulate; (c) a suspension in a suitable liquid; and (d) a suitable emulsion. Tablet forms may include one or more of lactose, mannitol, corn starch, potato starch, microcrystalline cellulose, acacia, gelatin, colloidal silicon dioxide, croscarmellose sodium, talc, magnesium stearate, stearic acid and other excipients, colorants, diluents, buffering agents, wetting agents, preservatives, flavoring agents and pharmacologically compatible excipients. Lozenge forms may include the active ingredient in a flavoring agent, typically sucrose and acacia or tragacanth, as well as pastilles comprising the active ingredient in an inert base such as gelatin and glycerin or sucrose and acacia, emulsions, gels and the like containing such excipients in addition to the active ingredient, as is known in the art.

Examples of suitable carriers, excipients, and diluents include, but are not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginates, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, saline solutions, syrups, methylcellulose, methyl-and propylhydroxybenzoates, talc, magnesium stearate, and mineral oil. The formulations may additionally include lubricating agents, wetting agents, emulsifying and suspending agents, preserving agents, sweetening agents, or flavoring agents.

Formulations suitable for parenteral administration, e.g., injection, include aqueous and non-aqueous, sterile isotonic injection solutions, which may contain antioxidants, buffers, bacteriostats, and solutes that render the formulation compatible with the blood of the intended recipient; and aqueous and non-aqueous sterile suspensions which may include suspending agents, solubilizers, thickeners, stabilizers, and preservatives. The formulations may be presented in unit-dose or multi-dose sealed containers, for example, ampoules and vials, and may be stored in a freeze-dried condition requiring only the addition of the sterile liquid excipient, for example water, for injections, immediately prior to use. Ready-to-use injection solutions and suspensions can be prepared from sterile powders, granules, and tablets of the kind previously described. Injectable formulations are preferred.

In some embodiments, the composition is formulated to have a pH range of about 4.5 to about 9.0, including, for example, any one of the following: about 5.0 to about 8.0, about 6.5 to about 7.5, and about 6.5 to about 7.0. In some embodiments, the pH of the composition is formulated to be no less than about 6, including, for example, no less than about any of the following: 6.5, 7 or 8 (e.g. about 8). The composition may also be made isotonic with blood by the addition of suitable tonicity adjusting agents such as glycerin.

Kit and device

The invention also provides kits and devices for use in any of the methods described herein.

For example, in some embodiments, a catheter with a needle (e.g., a deployable needle) is provided, wherein the needle comprises a composition comprising nanoparticles comprising a macrolide (e.g., rapamycin) and an albumin. In some embodiments, a catheter with a needle (e.g., a deployable needle) is provided, wherein the needle comprises a composition comprising nanoparticles comprising a macrolide (e.g., rapamycin) coated with albumin. In some embodiments, a catheter with a needle (e.g., a deployable needle) is provided, wherein the needle comprises a composition comprising nanoparticles comprising a macrolide (e.g., rapamycin) and an albumin, wherein the average particle size of the nanoparticles in the composition is no greater than about 200nm (such as less than about 200nm, for example, no greater than about 100 nm). In some embodiments, a catheter with a needle (e.g., a deployable needle) is provided, wherein the needle comprises a composition comprising nanoparticles comprising a macrolide (e.g., rapamycin) coated with albumin, wherein the average particle size of the nanoparticles in the composition is no greater than about 200nm (such as less than about 200nm, for example, no greater than about 100 nm). In some embodiments, a catheter with a needle (e.g., a deployable needle) is provided, wherein the needle comprises Nab-rapamycin. In some embodiments, the needle is encapsulated in the balloon. In some embodiments, the needle diameter is about 0.1 to about 3mm, including for example about 0.2 to about 2mm, about 0.5 to about 1mm, about 0.6 to about 0.9mm, or about 0.9 mm. The pin length is typically between about 20 and 3000 microns, including for example, between about 20-50, about 50-100, about 100-200, about 200-300, about 300-400, about 400-500, about 500-600, about 600-700, about 700-800, about 800-900, about 1-2, and about 2-3 microns. In some embodiments, the catheter comprises greater than 1 (e.g., 2, 3, 4, 5, 6,7, or more) needles.

Also provided are kits comprising one or more containers comprising a macrolide-containing nanoparticle composition (or unit dosage form and/or article of manufacture), and in some embodiments, further comprising instructions for use according to any of the methods described herein. In some embodiments, the kit includes a catheter with a needle that can be advanced from the lumen of the vessel, through the wall of the vessel (e.g., so that the needle hole is positioned outside the outer elastic layer of the wall); and a nanoparticle composition comprising a macrolide and an albumin, wherein the nanoparticle composition is injectable through a needle. In some embodiments, the kit further comprises a syringe. In some embodiments, the syringe is filled with an effective amount of the nanoparticle composition.

The kit may further comprise individual selection instructions suitable for treatment. The instructions provided in the kits of the invention are typically written 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 storage or optical disk) are also acceptable.

The kit of the invention is in suitable packaging. Suitable packaging includes, but is not limited to, vials, bottles, jars, flexible packaging (e.g., sealed Mylar or plastic bags), and the like. The kit may optionally provide additional components, such as buffers and explanatory information. The present application thus also provides articles of manufacture including vials (e.g., sealed vials), bottles, jars, flexible packaging, and the like.

Instructions for use of the nanoparticle compositions generally include information about the dosage, schedule of administration, and specific instructions for delivery of the nanoparticle composition. The container may be a unit dose, a bulk (e.g., multi-dose pack), or a sub-unit dose. Kits may also include multiple unit doses of macrolide and pharmaceutical composition and instructions for use, and packaged in sufficient quantities to be stored and used in pharmacies such as hospital pharmacies and compound pharmacies.

Those skilled in the art will appreciate that there are numerous embodiments within the scope and spirit of the present invention. The invention will now be described in more detail with reference to the following non-limiting examples. The following examples further illustrate the invention but, of course, should not be construed as in any way limiting its scope.

Examples

Example 1 periadventitial injection method Using micro-irrigation catheter

This example demonstrates Nab-rapamycin injected in periadventitial tissue. Nab-rapamycin (Celgene Corporation) was reconstituted in saline at 5mg/ml prior to injection.

For injection of Nab-rapamycin into periadventitial tissuesA micro-irrigation catheter (mercatormed systems, San leindro CA) was introduced into the artery while it was in a contracted state and a 0.9mm needle was covered in a balloon (fig. 1A). When the balloon is inflated, the needle is projected outward perpendicular to the catheter axis, at which point the backing (backing) balloon provides the opposite force to slide the needle into the artery wall (fig. 1B). Nab-rapamycin was then delivered through the needle into periadventitial tissues.

Example 2 periadventitial delivery of Nab-rapamycin in a porcine femoral artery balloon injury model

This experiment was performed to determine whether periadventitial delivery of Nab-rapamycin can reduce luminal stenosis in a porcine femoral balloon angioplasty injury model.

Sixteen young male Yorkshire hybrid pigs (average weight 34.9. + -. 2.3kg) were used in 2 study groups (FIG. 2). Percutaneous access is achieved through the carotid artery after general anesthesia. Animals were given intravenous heparin (5000 units). All pigs remained with 81mg aspirin daily after surgery. After sacrifice, the femoral artery was flushed with 1 liter of lactated Ringer's solution. The arteries were then harvested (pharmacokinetic group), or subsequently fixed by perfusion with 10% buffered formalin (perfusion fixed) at 120mmHg for 10 minutes and then harvested (histopathological group).

Nab-rapamycin was injected by periadventitial injection. Initial diagnostic angiography revealed a target femoral artery diameter of 4mm (fig. 3A). A micro-infusion catheter was placed in the mid-femoral artery and the balloon was inflated (fig. 3B). Periadventitial injections of Nab-rapamycin solution and 20% iodinated contrast agent (IsoVue 370) showed circumferential coverage of the vessels (fig. 3C-3E). The completion of the angiogram reveals the unobstructed femoral artery (fig. 3F). 32 injection sites were 100% surgically successful. The average injection time was 90 seconds (1 ml/min). No exfoliation, early or late thrombosis, bleeding or arteriovenous fistula.

Histomorphometry results were analyzed following periadventitial injection of Nab-rapamycin in the femoral artery. On day 28 post-treatment, the femoral artery treated with periadventitial Nab-rapamycin had a significantly increased luminal cross-sectional area p of 0.01(ANOVA) (fig. 4A), and a significantly increased total vessel cross-sectional area (fig. 4B), p of 0.005 (ANOVA). The percentage of luminal stenosis at day 28 had a tendency to decrease by Nab-rapamycin treatment. Luminal narrowing of femoral artery treated with periadventitial Nab-rapamycin (500 μ g) decreased by 42% on day 28 (19.5+ 3% vs 11.4+ 0.8%, p ═ 0.01 t-test) (fig. 4C). On day 28, mean medial fibrotic core of Nab-rapamycin treated arteries was significantly reduced compared to vehicle-only treated control arteries (fig. 4D), with p <0.0001 (ANOVA).

Pharmacokinetic results were analyzed after periadventitial injection of Nab-rapamycin in the femoral artery. Blood (serum) rapamycin levels were elevated within the first hour after a single periadventitial injection of Nab-rapamycin at 500 μ g, but declined by day 1 and could not be detected by day 28 (fig. 5A). In the femoral artery and peripheral perivascular tissues, rapamycin concentrations exceeded 1500-fold of serum concentrations at 1 hour. Rapamycin lasted 8 days and was not detectable by day 28 (fig. 5B).

Histopathological results after periadventitial injection of Nab-rapamycin in the femoral artery were analyzed. Representative femoral artery segments 28 days post treatment with vehicle (FIGS. 6A and 6B) or Nab-rapamycin 500 μ g (FIGS. 6C and 6D) are shown in FIG. 6. Nab-rapamycin treatment was associated with a significant reduction in medial fibrosis with similar levels of internal elastic layer injury (inset, 100 ×). The vessels were stained with H & E (fig. 6A and 6C) and trichromatic methods (fig. 6B and 6D) and imaged at 25 x.

Further histomorphometric analysis showed that Nab-rapamycin treatment was associated with a significantly reduced proliferation index (fig. 7A). On the other hand, endothelialization of the control and Nab-rapamycin treated femoral arteries was not different at day 28 (fig. 7B).

Further pharmacokinetic analysis showed that the proliferation index of balloon-injured arteries treated with 500 μ g Nab-rapamycin decreased significantly between days 3 and 28, with p ═ 0.004(ANOVA) (fig. 8A). Re-endothelialization occurred by day 8 (fig. 8B).

Furthermore, on day 3, adventitial leukocytes were significantly reduced in arteries treated with periadventitial Nab-rapamycin (fig. 9A). By day 28, Nab-rapamycin treated arteries had significantly reduced adventitial vessels (fig. 9B).

The results reported herein demonstrate that periadventitial delivery of Nab-rapamycin is associated with a temporary increase and a rapid decrease in serum rapamycin levels. Rapamycin levels in perivascular tissues exceeded 1500-fold higher than blood rapamycin levels at 1 hour post-treatment, and rapamycin remained in perivascular tissues for at least 8 days (fig. 5B). Balloon-injured femoral artery treated with Nab-rapamycin was significantly larger than vehicle-treated arteries, indicating less negative remodeling. In addition, periadventitial delivery of Nab-rapamycin resulted in a significant reduction in medial fibrosis.

Nab-rapamycin treated arteries demonstrated significantly reduced early (day 3) adventitial leukocyte infiltration. Ki-67 proliferation index of Nab-rapamycin treated arteries was significantly reduced at day 28 (FIG. 8A).

Endothelialization of the control and Nab-rapamycin treated femoral arteries was not different at day 28, and re-endothelialization of vessels following balloon-injured Nab-rapamycin treatment appeared to occur within the first week and appeared to be complete by day 8 (fig. 10). Nab-rapamycin treatment resulted in a significant decrease in proliferation and a significant decrease in medial fibrosis score, suggesting that rapamycin may affect the mechanism of balloon-damaged femoral remodeling.

Early reduction in outer membrane leukocyte infiltration and subsequent reduction in medial fibrosis and Ki-67 proliferation index at day 28 revealed a mechanism by which Nab-rapamycin can be effective around the outer membrane.

Although the foregoing invention has been described in some detail by way of illustration and example for purposes of clarity of understanding, it will be apparent to those skilled in the art that certain minor changes and modifications will be practiced. Therefore, the description and examples should not be construed as limiting the scope of the invention.

Claims (10)

1. A method of inhibiting negative remodeling in a blood vessel in an individual in need thereof, comprising injecting into the blood vessel wall or tissue surrounding the blood vessel wall an effective amount of a composition comprising nanoparticles comprising a macrolide and an albumin.

2. A method of inhibiting vascular fibrosis in a blood vessel of an individual in need thereof, comprising injecting into the blood vessel wall or tissue surrounding the blood vessel wall an effective amount of a composition comprising nanoparticles comprising a macrolide and an albumin.

3. The method of claim 2, wherein the vascular fibrosis is medial vascular fibrosis or adventitial fibrosis.

4. A method of promoting positive remodeling in a blood vessel in an individual in need thereof, comprising injecting into the blood vessel wall or tissue surrounding the blood vessel wall an effective amount of a composition comprising nanoparticles comprising a macrolide and an albumin.

5. The method of any one of claims 1-4, wherein the blood vessel is an artery.

6. The method of claim 5, wherein the artery is a coronary artery or a peripheral artery.

7. The method of claim 6, wherein the artery is selected from the group consisting of a renal artery, a cerebral artery, a pulmonary artery, and a leg artery.

8. The method of any one of claims 1-4, wherein the blood vessel is a vein.

9. The method of any one of claims 1-8, wherein the nanoparticle composition is injected into the vessel wall.

10. The method of any one of claims 1-8, wherein the nanoparticle composition is injected into the tissue surrounding the blood vessel wall.