Patent Application
20230131324
The present technology relates generally to formulations of antibodies to anti-endothelial lipase. The formulations are stable and reduce undesirable drug product characteristics, such as reversible self-association, precipitation and cloudiness. The formulations also have viscosities suitable for injection either manually or with automated injection systems.
Endothelial lipase is a phospholipase of the triglyceride lipase family that catalyzes the conversion of high-density lipoprotein (HDL) to low-density lipoprotein (LDL). Jaye et al., Nature Genetics 21:424-428 (1999). HDL is informally referred to as “good cholesterol” and has been shown to transport fat molecules out of artery walls, reduce macrophage accumulation, and therefore help prevent or even regress atherosclerosis (the narrowing of arteries). Humanized monoclonal antibodies inhibiting the activity of endothelial lipase have been developed, examples of which are described in US Application Publication No. 2017/0260290.
Increased plasma endothelial lipase concentrations have been associated with a deteriorated lipoprotein-lipid profile along with elevated plasma triglyceride and apolipoprotein B concentrations, as well as with smaller LDL particle size. (Paradis et al., Can J Cardiol 22: 31B-34B (2006).) Elevated proinflammatory cytokine concentrations and an increased prevalence of the metabolic syndrome have also been observed among individuals with elevated plasma endothelial lipase concentrations. (Id.) Given these and other factors, endothelial lipase has been considered to play an important role in cardiovascular disease. (Id.)
Despite the effectiveness of current therapies for cardiovascular disease such as high potency statins, there remains a significant residual risk of major adverse cardiovascular events in patients with acute coronary syndrome. The majority of myocardial infarctions occur in patients with normal LDL levels, and despite treatment with high dose and highly potent statins, PCSK9 inhibitors, and/or ezetimibe following a myocardial infarction, the residual risk of a second cardiovascular event remains high. For example, the IMPROVE-IT study (ezetimibe+statin) had a 33% risk of a cardiovascular event with 7 years of follow-up. In addition, ODYSSEY (PCSK9 inhibitor+statin) has a 9.5% residual risk over 2.8 years.
Several attempts to pharmacologically raise HDL levels using different mechanisms of action were found to not significantly improve cardiovascular outcomes. In particular, four trials of cholesterol transfer protein (CETP) inhibitors have been completed. While CETP inhibitors raise HDL cholesterol, three of the four trials did not improve cardiovascular outcomes, and in the one trial that did reduce cardiovascular events, the effect was modest with only a 9% relative risk reduction. This approach has been criticized since the inhibition of CETP causes a block in LDL receptor-mediated reverse cholesterol transport.
Humans with partial and complete loss of function mutations in the gene encoding endothelial lipase exhibit elevated HDL cholesterol, increased cholesterol efflux capacity, and trends towards reduced cardiovascular risk. Thus, neutralization of EL represents a promising therapeutic mechanism.
Biological pharmaceutical products, often called biologics, are frequently administered via injection. Solutions of large molecular weight biologics, such as monoclonal antibodies, typically have high concentrations and high viscosities that can lead to issues with product suitability and injection delivery. Particular suitability concerns for antibodies include reversable self-association (the formation of oligomeric species as a result of noncovalent intermolecular interactions), precipitation of protein and the osmolality and cloudiness of the solution. Viscosity and suitability can affect manufacturing, stability, delivery, and safety of the biologic. Thus, there is a need in the art for suitable formulations of biologics, such as anti-endothelial lipase antibodies, having suitable concentrations of active agent and viscosities suitable for injection.
The present disclosure is directed to a pharmaceutical formulation comprising: (i) about 40 mg/mL to about 160 mg/mL of an antibody that specifically binds to human endothelial lipase, and (ii) about 50 mM to about 260 mM arginine salt. In some embodiments, the pharmaceutical formulation comprises about 125 mg/mL of the antibody that specifically binds to endothelial lipase. In some embodiments, the pharmaceutical formulation comprises about 100 mg/mL of the antibody that specifically binds to endothelial lipase.
In some embodiments of the pharmaceutical formulation, the arginine salt comprises arginine hydrochloride, arginine acetate, arginine glutamate, or arginine sulfate. In some embodiments, the pharmaceutical formulation comprises arginine hydrochloride. In some embodiments, the pharmaceutical formulation comprises about 220 mM arginine hydrochloride. In some embodiments, the pharmaceutical formulation comprises about 160 mM arginine hydrochloride. In some embodiments, the pharmaceutical formulation comprises about 80 mM arginine hydrochloride.
In some embodiments of the pharmaceutical formulation, the antibody that specifically binds to endothelial lipase has a first heavy chain complementarity determining region (CDR1) with an amino acid sequence at least 80% identical to SEQ ID NO: 1. In some embodiments of the pharmaceutical formulation, the antibody that specifically binds to endothelial lipase has a heavy chain CDR1 comprising SEQ ID NO: 1. In some embodiments of the pharmaceutical formulation, the antibody that specifically binds to endothelial lipase has a second heavy chain complementarity determining region (CDR2) with an amino acid sequence at least 90% identical to SEQ ID NO: 2. In some embodiments of the pharmaceutical formulation, the antibody that specifically binds to endothelial lipase has a heavy chain CDR2 comprising SEQ ID NO:2. In some embodiments of the pharmaceutical formulation, the antibody that specifically binds to endothelial lipase has a third heavy chain complementarity determining region (CDR3) with an amino acid sequence at least 90% identical to SEQ ID NO: 3. In some embodiments of the pharmaceutical formulation, the antibody that specifically binds to endothelial lipase has a heavy chain CDR3 comprising SEQ ID NO:3.
In some embodiments of the pharmaceutical formulation, the antibody that specifically binds to endothelial lipase has a first light chain complementarity determining region (CDR1) with an amino acid sequence at least 90% identical to SEQ ID NO: 5. In some embodiments of the pharmaceutical formulation, the antibody that specifically binds to endothelial lipase has a light chain CDR1 comprising SEQ ID NO:5. In some embodiments of the pharmaceutical formulation, the antibody that specifically binds to endothelial lipase has a second light chain complementarity determining region (CDR2) with an amino acid sequence at least 80% identical to SEQ ID NO: 6. In some embodiments of the pharmaceutical formulation, the antibody that specifically binds to endothelial lipase has a light chain CDR2 comprising SEQ ID NO:6. In some embodiments of the pharmaceutical formulation, the antibody that specifically binds to endothelial lipase has a third light chain complementarity determining region (CDR3) with an amino acid sequence at least 80% identical to SEQ ID NO: 7. In some embodiments of the pharmaceutical formulation, the antibody that specifically binds to endothelial lipase has a light chain CDR3 comprising SEQ ID NO:7.
In some embodiments of the pharmaceutical formulation, the heavy chain of the antibody that specifically binds to endothelial lipase comprises an amino acid sequence at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 4. In some embodiments of the pharmaceutical formulation, the light chain of the antibody that specifically binds to endothelial lipase comprises an amino acid sequence at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 8. In some embodiments of the pharmaceutical formulation, the antibody that specifically binds to endothelial lipase comprises a heavy chain having an amino acid sequence comprising SEQ ID NO:4 and a light chain comprising an amino acid sequence comprising SEQ ID NO: 8.
In some embodiments of the pharmaceutical formulation, the antibody that specifically binds to endothelial lipase comprises a heavy chain comprising an amino acid sequence at least 90% identical to SEQ ID NO:9 and a light chain comprising an amino acid sequence at least 90% identical to SEQ ID NO:10. In some embodiments of the pharmaceutical formulation, the antibody that specifically binds to endothelial lipase comprises a heavy chain comprising SEQ ID NO:9 and a light chain comprising SEQ ID NO:10.
In some embodiments, the pharmaceutical formulation further comprises histidine. In some embodiments, the pharmaceutical formulation further comprises from about 5 mM to about 40 mM histidine. In some embodiments, the pharmaceutical formulation further comprises about 20 mM histidine. In some embodiments, the pharmaceutical formulation further comprises about 10 mM histidine. In some embodiments of the pharmaceutical formulation further comprising histidine, the histidine is a hydrochloride salt.
In some embodiments, the pharmaceutical formulation further comprises a surfactant. In some embodiments, the pharmaceutical formulation further comprises the surfactant polysorbate 20, polysorbate 60 or polysorbate 80. In some embodiments, the pharmaceutical formulation further comprises the surfactant polysorbate 80. In some embodiments, the pharmaceutical formulation further comprises from about 0.005% to about 0.05% polysorbate 80. In some embodiments, the pharmaceutical formulation further comprises about 0.01% polysorbate 80. In some embodiments, the pharmaceutical formulation further comprises about 0.02% polysorbate 80. In some embodiments, the pharmaceutical formulation further comprises about 0.04% polysorbate 80.
In some embodiments, the pharmaceutical formulation has a pH less than 7.0. In some embodiments, the pharmaceutical formulation has a pH of about 5.0 to about 7.0. In some embodiments, the pharmaceutical formulation has a pH of about 6.0. In some embodiments, the pharmaceutical formulation has a viscosity of less than 30 cP at 18° C. In some embodiments, the pharmaceutical formulation has an osmolality of about 260 to about 500 mOsm/kg.
In some embodiments, the pharmaceutical formulation is suitable for injection. In some embodiments, the pharmaceutical formulation is suitable for intravenous injection. In some embodiments, the pharmaceutical formulation is suitable for subcutaneous injection. In some embodiments, the pharmaceutical formulation is suitable for intramuscular injection.
In some embodiments, the pharmaceutical formulation is reconstituted from a lyophilized formulation. In some embodiments, the pharmaceutical formulation is a liquid formulation. In some embodiments, the pharmaceutical formulation is suitable for administration with an injection device such as an autoinjector device or a large bolus injector. In some embodiments, the pharmaceutical formulation is suitable for administration with an injection device such as a pre-filled syringe.
The present disclosure is also directed to methods comprising administering any of the pharmaceutical formulations disclosed herein. In some embodiments, the disclosure is directed to a method for treating cardiovascular disease in a mammalian subject in need thereof comprising administering to the subject any of the pharmaceutical formulations disclosed herein. In some embodiments, the disclosure is directed to a method for preventing cardiovascular disease in a mammalian subject in need thereof comprising administering to the subject any of the pharmaceutical formulations disclosed herein. In some embodiments, the disclosure is directed to a method for reducing atherosclerosis in a mammalian subject in need thereof comprising administering to the subject any of the pharmaceutical formulations disclosed herein. In some embodiments, the disclosure is directed to a method for reducing the risk of cardiovascular death, myocardial infarction, stroke, and/or coronary revascularization in a mammalian subject with prior acute coronary syndrome comprising administering to the subject any of the pharmaceutical formulations disclosed herein.
In some embodiments, the disclosure is directed to a method for preventing a secondary cardiovascular event in a mammalian subject in need thereof comprising administering to the subject any of the pharmaceutical formulations disclosed herein. In some embodiments, the disclosure is directed to a method for reducing the risk of a major adverse cardiovascular event in a mammalian subject in need thereof comprising administering to the subject any of the pharmaceutical formulations disclosed herein. In some embodiments, the disclosure is directed to a method for increasing the concentration of high-density lipoprotein in the blood of a mammalian subject in need thereof comprising administering to the subject any of the pharmaceutical formulations disclosed herein.
In some embodiments, the disclosure is directed to a method for increasing one or more clinical endpoints related to high-density lipoprotein in a mammalian subject in need thereof comprising administering to the subject any of the pharmaceutical formulations disclosed herein, wherein the clinical endpoints are HDL-c blood concentration, HDL particle number, HDL particle size, HDL phospholipid blood concentration, ApoA1 blood concentration and cholesterol efflux capacity. In some embodiments, the disclosure is directed to a method for decreasing inflammation in a mammalian subject in need thereof comprising administering to the subject any of the pharmaceutical formulations disclosed herein. In some embodiments, the disclosure is directed to a method for preventing inflammation in a mammalian subject in need thereof comprising administering to the subject any of the pharmaceutical formulations disclosed herein.
In embodiments of the methods disclosed herein, the pharmaceutical formulation is administered with an injection device. In embodiments of the methods disclosed herein, the pharmaceutical formulation is administered with an autoinjection device. In embodiments of the methods disclosed herein, the pharmaceutical formulation is administered with a large volume bolus injector. In embodiments of the methods disclosed herein, the pharmaceutical formulation is administered with a pre-filled syringe.
In embodiments of the methods disclosed herein, the mammalian subject is a human.
The present disclosure is further directed to a lyophilized pharmaceutical formulation comprising i) an antibody that specifically binds to endothelial lipase and ii) arginine hydrochloride; wherein the lyophilized pharmaceutical formulation can be reconstituted to a concentration of about 100 mg/mL to about 160 mg/mL of an antibody that specifically binds to endothelial lipase, and about 120 mM to about 260 mM arginine salt; and wherein the lyophilized pharmaceutical formulation is pharmaceutically acceptable.
The lyophilized pharmaceutical formulation comprising i) an antibody that specifically binds to endothelial lipase, ii) arginine hydrochloride, iii) histidine or a salt thereof, and iv) polysorbate 80; wherein the lyophilized pharmaceutical formulation can be reconstituted to a concentration of about 125 mg/mL antibody that specifically binds to endothelial lipase, about 160 mM arginine hydrochloride, about 20 mM histidine or a salt thereof and about 0.04% polysorbate 80; and wherein the lyophilized pharmaceutical formulation is pharmaceutically acceptable.
The present disclosure is further directed to a lyophilized pharmaceutical formulation comprising i) an antibody that specifically binds to endothelial lipase, ii) arginine hydrochloride, iii) histidine or a salt thereof, and iv) polysorbate 80; wherein the lyophilized pharmaceutical formulation can be reconstituted to a concentration of about 125 mg/mL antibody that specifically binds to endothelial lipase, about 220 mM arginine hydrochloride, about 20 mM histidine or a salt thereof and about 0.04% polysorbate 80; and wherein the lyophilized pharmaceutical formulation is pharmaceutically acceptable.
The present disclosure is further directed to a lyophilized pharmaceutical formulation comprising i) an antibody that specifically binds to endothelial lipase, ii) arginine hydrochloride, iii) histidine or a salt thereof, and iv) polysorbate 80; wherein the lyophilized pharmaceutical formulation can be reconstituted to a concentration of about 100 mg/mL antibody that specifically binds to endothelial lipase, about 160 mM arginine hydrochloride, about 20 mM histidine or a salt thereof and about 0.04% polysorbate 80; and wherein the lyophilized pharmaceutical formulation is pharmaceutically acceptable. In embodiments of the lyophilized pharmaceutical formulations disclosed herein, the lyophilized pharmaceutical formulation can be reconstituted to a pH of about 6.0.
The present disclosure is further directed to a liquid pharmaceutical formulation comprising about 125 mg/mL of an antibody that specifically binds to endothelial lipase, about 220 mM arginine hydrochloride, about 10 mM histidine or a salt thereof and about 0.02% polysorbate 80 and is pharmaceutically acceptable.
The present disclosure is further directed to a liquid pharmaceutical formulation comprising about 100 mg/mL of an antibody that specifically binds to endothelial lipase, about 220 mM arginine hydrochloride, about 10 mM histidine or a salt thereof and about 0.02% polysorbate 80 and is pharmaceutically acceptable.
The present disclosure is further directed to a liquid pharmaceutical formulation comprising about 125 mg/mL of an antibody that specifically binds to endothelial lipase, about 220 mM arginine hydrochloride, about 20 mM histidine or a salt thereof and about 0.02% polysorbate 80 and is pharmaceutically acceptable.
The present disclosure is further directed to a liquid pharmaceutical formulation comprising about 100 mg/mL of an antibody that specifically binds to endothelial lipase, about 220 mM arginine hydrochloride, about 20 mM histidine or a salt thereof and about 0.02% polysorbate 80 and is pharmaceutically acceptable.
The present disclosure is further directed to a liquid pharmaceutical formulation comprising about 50 mg/mL of an antibody that specifically binds to endothelial lipase, about 80 mM arginine hydrochloride, about 10 mM histidine or a salt thereof and about 0.02% polysorbate 80, and is pharmaceutically acceptable.
In embodiments of the liquid pharmaceutical formulations disclosed herein, the liquid pharmaceutical formulation has a pH of about 6.0.
The present disclosure provides pharmaceutical formulations comprising an antibody that specifically binds to human endothelial lipase and an arginine salt. The present disclosure also provides methods for using the pharmaceutical formulations disclosed herein.
It should be appreciated that the particular implementations shown and described herein are examples and are not intended to otherwise limit the scope of the application in any way.
The published patents, patent applications, websites, company names, and scientific literature referred to herein are hereby incorporated by reference in their entirety to the same extent as if each was specifically and individually indicated to be incorporated by reference. Any conflict from any reference cited herein and the specific teachings of this specification shall be resolved in favor of the latter. Likewise, any conflict from an art-understood definition of a word or phrase and a definition of the word or phrase as specifically taught in this specification shall be resolved in favor of the latter.
As used herein, “a” or “an” may mean one or more. As used herein, when used in conjunction with the word “comprising,” the words “a” or “an” may mean one or more than one. As used herein, “another” or “a further” may mean at least a second or more.
Throughout this application, the term “about” is used to indicate that a value includes the inherent variation of error for the method/device being employed to determine the value, or the variation that exists among the study subjects. Typically, the term “about” is meant to encompass approximately or less than 1%, 2%, 3%, 4%, 5%, 6%, 7% 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19% or 20% variability, depending on the situation.
The use of the term “or” in the claims is used to mean “and/or”, unless explicitly indicated to refer only to alternatives or the alternatives are mutually exclusive, although the disclosure supports a definition that refers to only alternatives and “and/or.”
As used herein, the terms “comprising” (and any variant or form of comprising, such as “comprise” and “comprises”), “having” (and any variant or form of having, such as “have” and “has”), “including” (and any variant or form of including, such as “includes” and “include”) or “containing” (and any variant or form of containing, such as “contains” and “contain”) are inclusive or open-ended and do not exclude additional, unrecited, elements or method steps. It is contemplated that any embodiment discussed in this specification can be implemented with respect to any method, system, host cells, expression vectors, and/or composition of the present disclosure. Furthermore, compositions, systems, host cells, and/or vectors of the present disclosure can be used to achieve methods and proteins of the present disclosure.
The use of the term “for example” and its corresponding abbreviation “e.g.” (whether italicized or not) means that the specific terms recited are representative examples and embodiments of the disclosure that are not intended to be limited to the specific examples referenced or cited unless explicitly stated otherwise.
Technical and scientific terms used herein have the meaning commonly understood by one of skill in the art to which the present application pertains, unless otherwise defined. Reference is made herein to various methodologies and materials known to those of skill in the art.
The term “antibody” as used herein can refer to a protein comprising at least two heavy chains and two light chains connected by disulfide bonds. The term “antibody” can include naturally occurring antibodies as well as all recombinant forms of antibodies, e.g., humanized antibodies, fully human antibodies and chimeric antibodies. Each heavy chain can comprise a heavy chain variable region (VH) and a heavy chain constant region (CH). Each light chain can comprise a light chain variable region (VL) and a light chain constant region (CL). The term “antibody”, however, can also include other types of antibodies such as single domain antibodies, heavy chain antibodies, i.e. antibodies only composed of one or more, in particular two heavy chains, and nanobodies, i.e. antibodies only composed of a single monomeric variable domain. Examples of fragments or derivatives of an antibody include (i) Fab fragments, monovalent fragments consisting of the variable region and the first constant domain of each the heavy and the light chain; (ii) F(ab)2 fragments, bivalent fragments comprising two Fab fragments linked by a disulfide bridge at the hinge region; (iii) Fd fragments consisting of the variable region and the first constant domain CHi of the heavy chain; (iv) Fv fragments consisting of the heavy chain and light chain variable region of a single arm of an antibody; (v) scFv fragments, Fv fragments consisting of a single polypeptide chain; (vi) (Fv)2 fragments consisting of two Fv fragments covalently linked together; (vii) a heavy chain variable domain; and (viii) multibodies consisting of a heavy chain variable region and a light chain variable region covalently linked together in such a manner that association of the heavy chain and light chain variable regions can only occur intermolecular but not intramolecular.
Endothelial lipase (sometimes abbreviated LIPG or EL) is a lipase enzyme that is secreted by vascular endothelial cells. Endothelial lipase plays a role in lipoprotein metabolism, cytokine expression and the regulation of lipid composition in cells. Yu, et al., Histology and Histopathology 33 (1): 1-10 (2018). Endotheial lipase is a phospholipase of the triglyceride lipase family that catalyzes the conversion of high-density lipoprotein (HDL) to low-density lipoprotein (LDL). Jaye et al., Nature Genetics 21:424-428 (1999). Endothelial lipase is thought to play a role in cardiovascular diseases such as atherosclerosis. Paradis, et al., The Canadian Journal of Cardiology. 22 Suppl B (Suppl B): 31B-34B (2006).
The amino acid sequences for human endothelial lipase is known in the art and the mature version of the protein (lacking the leader sequence) is provided herein as the sequence of SEQ ID NO:13.
The pharmaceutical formulations disclosed comprise an antibody that specifically binds to endothelial lipase. In embodiments, exemplary antibodies that may be used in the pharmaceutical formulations are those disclosed in US Published Patent Application 2017/0260290 to Naito et al., which is hereby incorporated by reference herein for its disclosure of these antibodies. In embodiments, the antibody that specifically binds endothelial lipase has the heavy chain amino acid sequence from antibody h55A1-S6 disclosed in US2017/0260290. In embodiments, the antibody that specifically binds endothelial lipase has the light chain amino acid sequence from antibody h55A1-F1 disclosed in US2017/0260290. In embodiments, the antibody that specifically binds endothelial lipase has the heavy chain amino acid sequence from antibody h55A1-S6 and the light chain amino acid sequence from antibody h55A1-F1 as disclosed in US2017/0260290. Specific antibody sequences referred to herein are taken from US2017/0260290 as indicated in the Sequence table below.
In embodiments, the antibody that specifically binds to endothelial lipase is MEDI5884, as described herein and in the examples below.
In embodiments, the antibody that specifically binds to endothelial lipase comprises the six CDRs of the MEDI5884 antibody listed as provided in Tables 1 and 2.
1The VH CDRs in Table 1 are determined according to Kabat.
2The VL CDRs in Table 2 are determined according to Kabat.
In embodiments, the antibody that specifically binds to endothelial lipase comprises the VH of the MEDI5884 antibody listed in Table 3.
In embodiments, the antibody that specifically binds to endothelial lipase comprises the VL of the MEDI5884 antibody listed in Table 4.
In embodiments, the antibody that specifically binds to endothelial lipase comprises the VH and the VL of the MEDI5884 antibody listed in Tables 3 and 4.
In embodiments, the antibody that specifically binds to endothelial lipase comprises the heavy chain sequence of the MEDI5884 antibody listed in Table 5.
In embodiments, the antibody that specifically binds to endothelial lipase comprises the light chain sequence of the MEDI5884 antibody listed in Table 6.
In embodiments, the antibody that specifically binds to endothelial lipase comprises the heavy chain sequence and the light chain sequence of the MEDI5884 antibody listed in Tables 5 and 6.
With respect to the heavy chain of the antibody that binds endothelial lipase, in embodiments, the heavy chain is a gamma heavy chain. The constant region of a human IgG4P heavy chain can comprise the following amino acid sequence:
With respect to the light chain of the antibody that binds endothelial lipase, in some aspects of the present disclosure, the light chain is a kappa light chain. The constant region of a human C kappa light chain can comprise the following amino acid sequence:
In embodiments, the antibody that specifically binds endothelial lipase has a first heavy chain complementarity determining region (CDR1) with an amino acid sequence at least 80% identical to SEQ ID NO: 1. In embodiments, the antibody that specifically binds endothelial lipase has a second heavy chain complementarity determining region (CDR2) with an amino acid sequence at least 80% identical to SEQ ID NO: 2. In embodiments, the antibody that specifically binds endothelial lipase has a third heavy chain complementarity determining region (CDR3) with an amino acid sequence at least 80% identical to SEQ ID NO: 3. In embodiments, the antibody that specifically binds endothelial lipase has two heavy chain complementarity determining regions at least 80% identical to two of SEQ ID NOs. 1-3. In embodiments, the antibody that specifically binds endothelial lipase has heavy chain complementarity determining regions at least 80% identical to each of SEQ ID NOs. 1-3. In embodiments, the antibody that specifically binds endothelial lipase has a heavy chain variable domain amino acid sequence at least 80% identical to SEQ ID NO: 4. In embodiments, the antibody that specifically binds endothelial lipase has a heavy chain amino acid sequence at least 80% identical to SEQ ID NO: 9.
In embodiments, the antibody that specifically binds endothelial lipase has a first heavy chain complementarity determining region (CDR1) with an amino acid sequence at least 90% identical to SEQ ID NO: 1. In embodiments, the antibody that specifically binds endothelial lipase has a second heavy chain complementarity determining region (CDR2) with an amino acid sequence at least 90% identical to SEQ ID NO: 2. In embodiments, the antibody that specifically binds endothelial lipase has a third heavy chain complementarity determining region (CDR3) with an amino acid sequence at least 90% identical to SEQ ID NO: 3. In embodiments, the antibody that specifically binds endothelial lipase has two heavy chain complementarity determining regions at least 90% identical to two of SEQ ID NOs. 1-3. In embodiments, the antibody that specifically binds endothelial lipase has heavy chain complementarity determining regions at least 90% identical to each of SEQ ID NOs. 1-3. In embodiments, the antibody that specifically binds endothelial lipase has a heavy chain variable domain amino acid sequence at least 90% identical to SEQ ID NO: 4. In embodiments, the antibody that specifically binds endothelial lipase has a heavy chain amino acid sequence at least 90% identical to SEQ ID NO: 9.
In embodiments, the antibody that specifically binds endothelial lipase has a heavy chain CDR1 with an amino acid sequence at least 95% identical to SEQ ID NO: 1. In embodiments, the antibody that specifically binds endothelial lipase has a heavy chain CDR2 with an amino acid sequence at least 95% identical to SEQ ID NO: 2. In embodiments, the antibody that specifically binds endothelial lipase has a heavy chain CDR3 with an amino acid sequence at least 95% identical to SEQ ID NO: 3. In embodiments, the antibody that specifically binds endothelial lipase has two heavy chain CDRs at least 95% identical to two of SEQ ID NOs. 1-3. In embodiments, the antibody that specifically binds endothelial lipase has heavy chain CDRs at least 95% identical to each of SEQ ID NOs. 1-3. In embodiments, the antibody that specifically binds endothelial lipase has a heavy chain variable domain amino acid sequence at least 95% identical to SEQ ID NO: 4. In embodiments, the antibody that specifically binds endothelial lipase has a heavy chain amino acid sequence at least 95% identical to SEQ ID NO: 9.
In embodiments, the antibody that specifically binds endothelial lipase has a heavy chain CDR1 with an amino acid sequence at least 98% identical to SEQ ID NO: 1. In embodiments, the antibody that specifically binds endothelial lipase has a heavy chain CDR2 with an amino acid sequence at least 98% identical to SEQ ID NO: 2. In embodiments, the antibody that specifically binds endothelial lipase has a heavy chain CDR3 with an amino acid sequence at least 98% identical to SEQ ID NO: 3. In embodiments, the antibody that specifically binds endothelial lipase has two heavy chain CDRs at least 98% identical to two of SEQ ID NOs. 1-3. In embodiments, the antibody that specifically binds endothelial lipase has heavy chain CDRs at least 98% identical to each of SEQ ID NOs. 1-3. In embodiments, the antibody that specifically binds endothelial lipase has a heavy chain variable domain amino acid sequence at least 98% identical to SEQ ID NO: 4. In embodiments, the antibody that specifically binds endothelial lipase has a heavy chain amino acid sequence at least 98% identical to SEQ ID NO: 9.
In embodiments, the antibody that specifically binds endothelial lipase has a heavy chain CDR1 with an amino acid sequence at least 99% identical to SEQ ID NO: 1. In embodiments, the antibody that specifically binds endothelial lipase has a heavy chain CDR2 with an amino acid sequence at least 99% identical to SEQ ID NO: 2. In embodiments, the antibody that specifically binds endothelial lipase has a heavy chain CDR3 with an amino acid sequence at least 99% identical to SEQ ID NO: 3. In embodiments, the antibody that specifically binds endothelial lipase has two heavy chain CDRs at least 99% identical to two of SEQ ID NOs. 1-3. In embodiments, the antibody that specifically binds endothelial lipase has heavy chain CDRs at least 99% identical to each of SEQ ID NOs. 1-3. In embodiments, the antibody that specifically binds endothelial lipase has a heavy chain variable domain amino acid sequence at least 99% identical to SEQ ID NO: 4. In embodiments, the antibody that specifically binds endothelial lipase has a heavy chain amino acid sequence at least 99% identical to SEQ ID NO: 9.
In embodiments, the antibody that specifically binds endothelial lipase has a heavy chain CDR1 with an amino acid sequence comprising SEQ ID NO: 1. In embodiments, the antibody that specifically binds endothelial lipase has a heavy chain CDR2 with an amino acid sequence comprising SEQ ID NO: 2. In embodiments, the antibody that specifically binds endothelial lipase has a heavy chain CDR3 with an amino acid sequence comprising SEQ ID NO: 3. In embodiments, the antibody that specifically binds endothelial lipase has two heavy chain CDRs comprising two of SEQ ID NOs. 1-3. In embodiments, the antibody that specifically binds endothelial lipase has heavy chain CDRs comprising each of SEQ ID NOs. 1-3. In embodiments, the antibody that specifically binds endothelial lipase has a heavy chain variable domain amino acid sequence comprising SEQ ID NO: 4. In embodiments, the antibody that specifically binds endothelial lipase has a heavy chain amino acid sequence comprising SEQ ID NO: 9.
In embodiments, the antibody that specifically binds endothelial lipase has a light chain CDR1 with an amino acid sequence at least 80% identical to SEQ ID NO: 5. In embodiments, the antibody that specifically binds endothelial lipase has a light chain CDR2 with an amino acid sequence at least 80% identical to SEQ ID NO: 6. In embodiments, the antibody that specifically binds endothelial lipase has a light chain CDR3 with an amino acid sequence at least 80% identical to SEQ ID NO: 7. In embodiments, the antibody that specifically binds endothelial lipase has two light chain CDRs at least 80% identical to two of SEQ ID NOs. 5-7. In embodiments, the antibody that specifically binds endothelial lipase has light chain CDRs at least 80% identical to each of SEQ ID NOs. 5-7. In embodiments, the antibody that specifically binds endothelial lipase has a light chain variable domain amino acid sequence at least 80% identical to SEQ ID NO: 8. In embodiments, the antibody that specifically binds endothelial lipase has a light chain amino acid sequence at least 80% identical to SEQ ID NO: 10.
In embodiments, the antibody that specifically binds endothelial lipase has a light chain CDR1 with an amino acid sequence at least 90% identical to SEQ ID NO: 5. In embodiments, the antibody that specifically binds endothelial lipase has a light chain CDR2 with an amino acid sequence at least 90% identical to SEQ ID NO: 6. In embodiments, the antibody that specifically binds endothelial lipase has a light chain CDR3 with an amino acid sequence at least 90% identical to SEQ ID NO: 7. In embodiments, the antibody that specifically binds endothelial lipase has two light chain CDRs at least 90% identical to two of SEQ ID NOs. 5-7. In embodiments, the antibody that specifically binds endothelial lipase has light chain CDRs at least 90% identical to each of SEQ ID NOs. 5-7. In embodiments, the antibody that specifically binds endothelial lipase has a light chain variable domain amino acid sequence at least 90% identical to SEQ ID NO: 8. In embodiments, the antibody that specifically binds endothelial lipase has a light chain amino acid sequence at least 90% identical to SEQ ID NO: 10.
In embodiments, the antibody that specifically binds endothelial lipase has a light chain CDR1 with an amino acid sequence at least 95% identical to SEQ ID NO: 5. In embodiments, the antibody that specifically binds endothelial lipase has a light chain CDR2 with an amino acid sequence at least 95% identical to SEQ ID NO: 6. In embodiments, the antibody that specifically binds endothelial lipase has a light chain CDR3 with an amino acid sequence at least 95% identical to SEQ ID NO: 7. In embodiments, the antibody that specifically binds endothelial lipase has two light chain CDRs at least 95% identical to two of SEQ ID NOs. 5-7. In embodiments, the antibody that specifically binds endothelial lipase has light chain CDRs at least 95% identical to each of SEQ ID NOs. 5-7. In embodiments, the antibody that specifically binds endothelial lipase has a light chain variable domain amino acid sequence at least 95% identical to SEQ ID NO: 8. In embodiments, the antibody that specifically binds endothelial lipase has a light chain amino acid sequence at least 95% identical to SEQ ID NO: 10.
In embodiments, the antibody that specifically binds endothelial lipase has a light chain CDR1 with an amino acid sequence at least 98% identical to SEQ ID NO: 5. In embodiments, the antibody that specifically binds endothelial lipase has a light chain CDR2 with an amino acid sequence at least 98% identical to SEQ ID NO: 6. In embodiments, the antibody that specifically binds endothelial lipase has a light chain CDR3 with an amino acid sequence at least 98% identical to SEQ ID NO: 7. In embodiments, the antibody that specifically binds endothelial lipase has two light chain CDRs at least 98% identical to two of SEQ ID NOs. 5-7. In embodiments, the antibody that specifically binds endothelial lipase has light chain CDRs at least 98% identical to each of SEQ ID NOs. 5-7. In embodiments, the antibody that specifically binds endothelial lipase has a light chain variable domain amino acid sequence at least 98% identical to SEQ ID NO: 8. In embodiments, the antibody that specifically binds endothelial lipase has a light chain amino acid sequence at least 98% identical to SEQ ID NO: 10.
In embodiments, the antibody that specifically binds endothelial lipase has a light chain CDR1 with an amino acid sequence at least 99% identical to SEQ ID NO: 5. In embodiments, the antibody that specifically binds endothelial lipase has a light chain CDR2 with an amino acid sequence at least 99% identical to SEQ ID NO: 6. In embodiments, the antibody that specifically binds endothelial lipase has a light chain CDR3 with an amino acid sequence at least 99% identical to SEQ ID NO: 7. In embodiments, the antibody that specifically binds endothelial lipase has two light chain CDRs at least 99% identical to two of SEQ ID NOs. 5-7. In embodiments, the antibody that specifically binds endothelial lipase has light chain CDRs at least 99% identical to each of SEQ ID NOs. 5-7. In embodiments, the antibody that specifically binds endothelial lipase has a light chain variable domain amino acid sequence at least 99% identical to SEQ ID NO: 8. In embodiments, the antibody that specifically binds endothelial lipase has a light chain amino acid sequence at least 99% identical to SEQ ID NO: 10.
In embodiments, the antibody that specifically binds endothelial lipase has a light chain CDR1 with an amino acid sequence comprising SEQ ID NO: 5. In embodiments, the antibody that specifically binds endothelial lipase has a light chain CDR2 with an amino acid sequence comprising SEQ ID NO: 6. In embodiments, the antibody that specifically binds endothelial lipase has a light chain CDR3 with an amino acid sequence comprising SEQ ID NO: 7. In embodiments, the antibody that specifically binds endothelial lipase has two light chain CDRs comprising two of SEQ ID NOs. 5-7. In embodiments, the antibody that specifically binds endothelial lipase has light chain CDRs comprising each of SEQ ID NOs. 5-7. In embodiments, the antibody that specifically binds endothelial lipase has a light chain variable domain amino acid sequence comprising SEQ ID NO: 8. In embodiments, the antibody that specifically binds endothelial lipase has a light chain amino acid sequence comprising SEQ ID NO: 10.
In embodiments, the antibody that specifically binds endothelial lipase comprises one of more complementarity determining regions selected from SEQ ID NOs: 1-3 and 5-7. In embodiments, the antibody that specifically binds endothelial lipase has a heavy chain variable domain amino acid sequence comprising SEQ ID NO: 4 and a light chain variable domain amino acid sequence comprising SEQ ID NO: 8.
In embodiments, the concentration of the antibody that specially binds endothelial lipase in the pharmaceutical formulation is from about 40 mg/mL and about 160 mg/mL. In embodiments, the concentration of the antibody in the pharmaceutical formulation is from about 100 mg/mL and about 150 mg/mL. In embodiments, the concentration of the antibody in the pharmaceutical formulation is from about 110 mg/mL and about 140 mg/mL. In embodiments, the concentration of the antibody in the pharmaceutical formulation is from about 120 mg/mL and about 130 mg/mL. In embodiments, the concentration of the antibody in the pharmaceutical formulation is from about 40 mg/mL and about 60 mg/mL. In embodiments, the concentration of the antibody in the pharmaceutical formulation is from about 90 mg/mL and about 110 mg/mL. In embodiments, the concentration of the antibody in the pharmaceutical formulation is about 125 mg/mL. In embodiments, the concentration of the antibody in the pharmaceutical formulation is about 50 mg/mL. In embodiments, the concentration of the antibody in the pharmaceutical formulation is about 100 mg/mL.
The pharmaceutical formulations disclosed provide a high enough concentration of antibody specific to endothelial lipase to be effective in a low volume suitable for injection. In embodiments, the pharmaceutical formulations provide a high enough concentration of antibody specific to endothelial lipase to be effective at a viscosity suitable for injection. In embodiments, the pharmaceutical formulations provide a high enough concentration of antibody specific to endothelial lipase to be effective while avoiding undesirable reversible self-association (RSA). In embodiments, the pharmaceutical formulations provide a high enough concentration of antibody specific to endothelial lipase to be effective at an osmolality suitable for injection. In embodiments, the pharmaceutical formulations provide a high enough concentration of antibody specific to endothelial lipase to be effective while being storage stable.
It is frequently a challenge when formulating injectable antibodies to be able to formulate the antibody at concentration high enough to allow for effective dosing in a volume suitable for injection. Increasing the concentration of antibody in a formulation can lead to undesirable increases in viscosity, RSA, precipitation and cloudiness that make the formulation unsuitable for injection into a subject.
The pharmaceutical formulations disclosed herein comprise an arginine (Arg) salt as a viscosity regulating agent. In embodiments, the arginine salt is arginine hydrochloride, arginine acetate, arginine glutamate or arginine sulfate. In embodiments, the arginine salt is arginine hydrochloride. In embodiments, the pharmaceutical formulations may comprise more than one arginine salt, for example, mixtures of two or more of arginine hydrochloride, arginine acetate, arginine glutamate and arginine sulfate
In embodiments, the concentration of the arginine salt in the pharmaceutical formulation is from about 50 mM to about 260 mM. In embodiments, the concentration of the arginine salt in the pharmaceutical formulation is from about 80 mM to about 260 mM. In embodiments, the concentration of the arginine salt in the pharmaceutical formulation is from about 160 mM to about 240 mM. In embodiments, the concentration of the arginine salt in the pharmaceutical formulation is from about 180 mM to about 230 mM. In embodiments, the concentration of the arginine salt in the pharmaceutical formulation is from about 200 mM to about 225 mM. In embodiments, the concentration of the arginine salt in the pharmaceutical formulation is from about 110 mM to about 140 mM. In embodiments, the concentration of the arginine salt in the pharmaceutical formulation is from about 50 mM to about 110 mM. In embodiments, the concentration of the arginine salt in the pharmaceutical formulation is from about 60 mM to about 100 mM. In embodiments, the concentration of the arginine salt in the pharmaceutical formulation is from about 70 mM to about 90 mM. In embodiments, the concentration of the arginine salt in the pharmaceutical formulation is about 220 mM. In embodiments, the concentration of the arginine salt in the pharmaceutical formulation is about 160 mM. In embodiments, the concentration of the arginine salt in the pharmaceutical formulation is about 80 mM.
In embodiments, the pharmaceutical formulation has a concentration of from about 50 mM to about 260 mM arginine hydrochloride (ArgHCl). In embodiments, the pharmaceutical formulation has a concentration of from about 80 mM to about 260 mM arginine hydrochloride. In embodiments, the pharmaceutical formulation has a concentration of from about 160 mM to about 240 mM arginine hydrochloride. In embodiments, the pharmaceutical formulation has a concentration of from about 180 mM to about 230 mM arginine hydrochloride. In embodiments, the pharmaceutical formulation has a concentration of from about 200 mM to about 225 mM arginine hydrochloride. In embodiments, the pharmaceutical formulation has a concentration of from about 110 mM to about 140 mM arginine hydrochloride (ArgHCl). In embodiments, the pharmaceutical formulation has a concentration of from about 50 mM to about 110 mM arginine hydrochloride. In embodiments, the pharmaceutical formulation has a concentration of from about 60 mM to about 100 mM arginine hydrochloride. In embodiments, the pharmaceutical formulation has a concentration of from about 70 mM to about 90 mM arginine hydrochloride. In embodiments, the pharmaceutical formulation has a concentration of about 220 mM arginine hydrochloride. In embodiments, the pharmaceutical formulation has a concentration of about 160 mM arginine hydrochloride. In embodiments, the pharmaceutical formulation has a concentration of about 80 mM arginine hydrochloride.
In embodiments, the pharmaceutical formulations further comprise histidine (His) as a buffer. In embodiments, the histidine in the pharmaceutical formulation may be in base form or salt form, or in combinations thereof. In embodiments where the histidine is in salt form, the salt form may be histidine hydrochloride, histidine acetate, histidine or histidine sulfate, or combinations thereof. In embodiments, the pharmaceutical formulation comprises histidine hydrochloride (HisHCl) In embodiments, the pharmaceutical formulation has a concentration of from about 5 mM to about 40 mM histidine and/or histidine salt. In embodiments, the pharmaceutical formulation has a concentration of about 10 mM histidine and/or histidine salt. In embodiments, the pharmaceutical formulation has a concentration of about 20 mM histidine and/or histidine salt.
In embodiments, the pharmaceutical formulations comprise other buffers such as acetate and phosphate buffers. In embodiments, the pharmaceutical formulation has a concentration of from about 5 mM to about 40 mM buffer. In embodiments, the pharmaceutical formulation has a concentration of about 10 mM buffer. In embodiments, the pharmaceutical formulation has a concentration of about 20 mM buffer.
In embodiments, the pharmaceutical formulations further comprise a surfactant. In embodiments, the surfactant is polysorbate 20, polysorbate 60, polysorbate 80 or combinations thereof. In embodiments, the surfactant is polysorbate 80 (PS80). In embodiments, the pharmaceutical formulation has a concentration of from about 0.005% to about 0.05% surfactant. In embodiments, the pharmaceutical formulation has a concentration of from about 0.02% to about 0.04% surfactant. In embodiments, the pharmaceutical formulation has a concentration of about 0.02% surfactant. In embodiments, the pharmaceutical formulation has a concentration of about 0.04% surfactant. In embodiments, the pharmaceutical formulation has a concentration of about 0.01% surfactant. In embodiments, the pharmaceutical formulation has a concentration of from about 0.01% to about 0.05% PS80. In embodiments, the pharmaceutical formulation has a concentration of from about 0.02% to about 0.04% PS80. In embodiments, the pharmaceutical formulation has a concentration of about 0.02% PS80. In embodiments, the pharmaceutical formulation has a concentration of about 0.04% PS80. In embodiments, the pharmaceutical formulation has a concentration of about 0.01% PS80.
In embodiments, the pharmaceutical formulations can further comprise additional excipients. In embodiments, the pharmaceutical formulation further comprises excipients suitable for use in injectable formulations, e.g., buffers, viscosity regulating agents, salts and stabilizers. In embodiments, the pharmaceutical formulation further comprises one or more of lysine, lysine hydrochloride, proline, glutamic acid, sodium sulfate, sodium chloride or dimethyl sulfoxide (DMSO).
In embodiments, the pharmaceutical formulations have a pH of less than 7.0. In embodiments, the pharmaceutical formulations have a pH of from about 5.0 to about 7.0. In embodiments, the pharmaceutical formulations have a pH of about 6.0.
The pharmaceutical formulations are designed to have a viscosity suitable for injection, e.g., with either an autoinjection of manual injection device. A formulation with too high of a viscosity requires greater injection force may not flow properly through the injection device either at all or only with the use of an undesirable large bore needle. Viscosity of such formulations is measured using methods known in the art as described further in the examples. As viscosity changes with temperature, it is usually measured at a common temperature, for example, 18° C. In embodiments, the pharmaceutical formulation has a viscosity of less than or equal to 30 centipoise (cP) at 18° C., i.e., about 1 to about 30 cP, about 2 to about 28 cP, about 3 to about 26 cP, about 5 to about 25 cP, about 6 to about 22 cP, about 7 to about 20 cP, about 8 to about 20 cP, about 9 to about 20 cP, about 10 to about 20 cP, or about 12 to about 18 cP at 18° C. In embodiments, the pharmaceutical formulation has a viscosity of less than or equal to 20 centipoise (cP) at 18° C. In embodiments, the pharmaceutical formulation has a viscosity of less than or equal to 15 centipoise (cP) at 18° C.
The pharmaceutical formulations are designed to have a volume suitable for injection with either an autoinjector or manual injector. In embodiments, the pharmaceutical formulation can have a volume of about 0.1 to about 4.0 mL, about 0.5 to about 3.5 mL, about 1.0 to about 3.0 mL, or about 1.5 to about 2.0 mL. In embodiments, the pharmaceutical formulation can have an injection volume of about 0.1 mL, about 0.25 mL, about 0.5 mL, about 0.75 mL, about 1.0 mL, about 1.25 mL, about 1.5 mL, about 1.75 mL, about 2.0 mL, about 2.25 mL, about 2.5, mL, about 2.75 mL, about 3.0 mL, about 3.25 mL, about 3.5 mL, about 3.75 mL or about 4.0 mL.
The pharmaceutical formulations can also be designed to prevent reversable self-association (RSA) between the antibody molecules in the formulation. RSA includes native, non-covalent, and reversible oligomerization of monomeric species of antibody or other protein. RSA is more likely in formulations having a high concentration of antibody because of the reduced intermolecular distance between individual molecules and the increased probability of molecular collisions. RSA can pose manufacturing and delivery challenges and can cause pain upon administration. RSA can be measured using techniques known in the art such as dynamic light scattering, analytical ultracentrifugation, differential scanning calorimetry, differential scanning fluorimetry and high performance size-exclusion chromatography.
The pharmaceutical formulations can also be designed to have an osmolality suitable for injection. The osmolality of an injectable formulation can be associated with pain upon injection and tolerability. Specific instruments and techniques for measuring osmolality are available in the art and described in the examples below. In embodiments, the pharmaceutical formulation has an osmolality of from about 260 mOsm/kg to about 500 mOsm/kg, i.e. from about 280 mOsm/kg to about 490 mOsm/kg, from about 320 mOsm/kg to about 480 mOsm/kg, from about 340 mOsm/kg to about 470 mOsm/kg, from about 360 mOsm/kg to about 460 mOsm/kg, from about 380 mOsm/kg to about 460 mOsm/kg, from about 400 mOsm/kg to about 460 mOsm/kg and from about 440 mOsm/kg to about 460 mOsm/kg. In embodiments, the pharmaceutical formulation has an osmolality of from about 440 mOsm/kg to about 460 mOsm/kg.
The pharmaceutical formulations can be designed to be suitable for injection to a subject and are suitable for use in injection systems known in the art. In embodiments, the pharmaceutical formulation is suitable for intravenous injection. In embodiments, the pharmaceutical formulation is suitable for subcutaneous injection. In embodiments, the pharmaceutical formulation is suitable for injection with an injection device. In embodiments, the pharmaceutical formulation is suitable for injection with an autoinjection device. In embodiments, the pharmaceutical formulation is suitable for injection with a large volume bolus injector. In embodiments, the pharmaceutical formulation is suitable for injection with a pre-filled syringe.
In embodiments, the pharmaceutical formulation is reconstituted from a lyophilized formulation. Liquids containing the formulation to be lyophilized can be lyophilized according to standard methods in the art. The lyophilized version of the formulation may be referred to as a drug product (DP) formulation. In embodiments, the lyophilized DP is reconstituted using the same volume of liquid present prior to lyophilization. In embodiments, the lyophilized DP is reconstituted using half the volume of liquid present prior to lyophilization (a half reconstitution or half recon). In a half reconstitution, the concentration of each component following reconstitution is then double the concentration of these components in the liquid prior to lyophilization. In embodiments, the lyophilized DP can be reconstituted with less volume than the original liquid. In embodiments, the lyophilized DP can be reconstituted with more volume than the original liquid.
In embodiments, the pharmaceutical formulation is a liquid formulation. The liquid version of the formulation may be referred to as a drug substance (DS) formulation or a liquid drug product (DP) formulation.
The pharmaceutical formulations can be designed to be storage stable. In embodiments, the pharmaceutical formulation is storage stable in lyophilized form. In embodiments, the pharmaceutical formulation is storage stable in liquid form. In embodiments, the pharmaceutical formulation is storage stable at a temperature of between about −80° C. to about −10° C. for a period of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 48 or 60 months. In embodiments, the pharmaceutical formulation is storage stable at a temperature of between about 16° C. to about 22° C. for a period of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or 18 months.
In embodiments, the pharmaceutical formulation is a lyophilized pharmaceutical formulation comprising an antibody that specifically binds to endothelial lipase and arginine hydrochloride, wherein the lyophilized pharmaceutical formulation can be reconstituted to a concentration of about 100 mg/mL to about 160 mg/mL of an antibody that specifically binds to endothelial lipase, and about 120 mM to about 260 mM arginine salt.
In embodiments, the pharmaceutical formulation is a lyophilized pharmaceutical formulation comprising an antibody that specifically binds to endothelial lipase, arginine hydrochloride, histidine or a salt thereof, and polysorbate 80, wherein the lyophilized pharmaceutical formulation can be reconstituted to a concentration of about 125 mg/mL antibody that specifically binds to endothelial lipase, about 160 mM arginine hydrochloride, about 20 mM histidine or a salt thereof and about 0.04% polysorbate 80.
In embodiments, the pharmaceutical formulation is a lyophilized pharmaceutical formulation comprising an antibody that specifically binds to endothelial lipase, arginine hydrochloride, histidine or a salt thereof, and polysorbate 80, wherein the lyophilized pharmaceutical formulation can be reconstituted to a concentration of about 125 mg/mL antibody that specifically binds to endothelial lipase, about 220 mM arginine hydrochloride, about 20 mM histidine or a salt thereof and about 0.04% polysorbate 80.
In embodiments, the pharmaceutical formulation is a lyophilized pharmaceutical formulation comprising an antibody that specifically binds to endothelial lipase, arginine hydrochloride, histidine or a salt thereof, and polysorbate 80, wherein the lyophilized pharmaceutical formulation can be reconstituted to a concentration of about 100 mg/mL antibody that specifically binds to endothelial lipase, about 160 mM arginine hydrochloride, about 20 mM histidine or a salt thereof and about 0.04% polysorbate 80.
In embodiments, the pharmaceutical formulation is a liquid pharmaceutical formulation comprising about 125 mg/mL of an antibody that specifically binds to endothelial lipase, about 220 mM arginine hydrochloride, about 10 mM histidine or a salt thereof and about 0.02% polysorbate 80.
In embodiments, the pharmaceutical formulation is a liquid pharmaceutical formulation comprising about 100 mg/mL of an antibody that specifically binds to endothelial lipase, about 220 mM arginine hydrochloride, about 10 mM histidine or a salt thereof and about 0.02% polysorbate 80.
In embodiments, the pharmaceutical formulation is a liquid pharmaceutical formulation comprising about 125 mg/mL of an antibody that specifically binds to endothelial lipase, about 220 mM arginine hydrochloride, about 20 mM histidine or a salt thereof and about 0.02% polysorbate 80.
In embodiments, the pharmaceutical formulation is a liquid pharmaceutical formulation comprising about 100 mg/mL of an antibody that specifically binds to endothelial lipase, about 220 mM arginine hydrochloride, about 20 mM histidine or a salt thereof and about 0.02% polysorbate 80.
In embodiments, the pharmaceutical formulation is a liquid pharmaceutical formulation comprising about 50 mg/mL of an antibody that specifically binds to endothelial lipase, about 80 mM arginine hydrochloride, about 10 mM histidine or a salt thereof and about 0.02% polysorbate 80.
The present disclosure is also directed to methods comprising administering any of the pharmaceutical formulations disclosed herein. The methods described involve inhibition of a function of endothelial lipase by administration of a pharmaceutical formulation comprising an antibody that specifically binds to endothelial lipase. In embodiments, the methods comprise administering a pharmaceutical formulation comprising an antibody that specifically binds to endothelial lipase and an arginine salt.
In embodiments, the pharmaceutical formulation is used in a method for treating cardiovascular disease in a mammalian subject in need thereof. In embodiments, the pharmaceutical formulation is used in a method for preventing cardiovascular disease in a mammalian subject in need thereof. In embodiments, the pharmaceutical formulation is used in a method for reducing atherosclerosis in a mammalian subject in need thereof. In embodiments, the pharmaceutical formulation is used in a method for reducing the risk of cardiovascular death, myocardial infarction, stroke, and/or coronary revascularization in a mammalian subject with prior acute coronary syndrome.
In embodiments, the pharmaceutical formulation is used in a method for preventing a secondary cardiovascular event in a mammalian subject in need thereof. In embodiments, the pharmaceutical formulation is used in a method for reducing the risk of a major adverse cardiovascular event in a mammalian subject in need thereof. In embodiments, the pharmaceutical formulation is used in a method for increasing the concentration of high-density lipoprotein in the blood of a mammalian subject in need thereof.
In embodiments, the pharmaceutical formulation is used in a method for increasing one or more clinical endpoints related to high-density lipoprotein in a mammalian subject in need thereof. In this embodiment, the clinical endpoints can be one or more of HDL-c blood concentration, HDL particle number, HDL particle size, HDL phospholipid blood concentration, ApoA1 blood concentration and cholesterol efflux capacity. In embodiments, the pharmaceutical formulation is used in a method for decreasing inflammation in a mammalian subject in need thereof. In embodiments, the pharmaceutical formulation is used in a method for preventing inflammation in a mammalian subject in need thereof.
The methods embodied herein may be practiced using an injection device, including an autoinjection device as described above. In embodiments, the methods embodied herein may be practiced using a large volume bolus injector or a pre-filled syringe. In embodiments, the pre-filled syringe is an accessorized pre-filled syringe having one or more safety feature, e.g., a retracting needle after injection, and/or one or more ergonomics features, e.g., a finger flange or other accessory that assists in holding the syringe.
In embodiments, doses of the antibody that specifically binds to endothelial lipase for the methods embodied herein range from 50 mg to 400 mg per month. In embodiments, doses of the antibody that specifically binds to endothelial lipase for the methods embodied herein range from 30 mg to 500 mg per month. In embodiments, the dose of antibody is from 100 mg to 350 mg per month. In embodiments, the dose of antibody is from 250 mg to 350 mg per month. In embodiments, the dose of antibody is about 250 mg per month. In embodiments, the dose of antibody is 250 mg per month. In embodiments, the dose of antibody is about 125 mg per month. In embodiments, the dose of antibody is 125 mg per month. One skilled in the art will understand that the pharmaceutical formulation may need to be administered multiple times or at increased volumes to achieve the desired dose. For example, for a pharmaceutical formulation with an antibody concentration of 125 mg/mL may administered with 2 mL in a single injection or via two injections of 1 mL in order to achieve a dose of 250 mg. One skilled in the art will also understand that increasing the dosing frequency, such as increasing from monthly to bi-weekly, will allow for multiple injections to achieve a higher total dose.
In embodiments of the methods embodied herein, the mammalian subject is a human. In embodiments of the methods embodied herein, the mammalian subject is an animal such as an agricultural animal (e.g., cattle, sheep, swine), research animal (e.g., mice, rats, monkeys, chimpanzees) or companion animal (e.g., dogs, cats and rabbits).
MEDI5884 is an IgG4P monoclonal antibody directed against endothelial lipase enzyme (SEQ ID NO: 13) involved in conversion of high density lipoprotein (HDL) to low density lipoprotein (LDL). The term “MEDI5884” refers to anti-EL antibody that comprises the heavy chain of SEQ ID NO:9 and the light chain of SEQ ID NO:10. MEDI5884 is also referred to as “S6F1-4P,” and it comprises the heavy chain variable region of the h55A1-S6 antibody and the light chain variable region of the h55A1-F1 antibody, which are disclosed in US Published Application No. 2017/0260290, which is herein incorporated by reference in its entirety. MEDI5884 has an overall molecular weight of approximately 150 kDa. The observed intact mass and peptide map are consistent with the expected primary structure, based on the DNA sequence. The heavy chain variable region of MEDI5884 is provided below as SEQ ID NO:4. The light chain variable region of MEDI5884 is provided below as SEQ ID NO:8. The full length heavy chain of MEDI5884 is provided below as SEQ ID NO:9. The full length light chain of MEDI5884 is provided below as SEQ ID NO:10. The experimentally determined extinction coefficient is 1.44 (mg/mL)−1cm−1 and the isoelectric point (pI) is in the range of 8.4-8.9. The first thermal transition as determined by differential scanning calorimetry is 56.4° C.
MEDI5884 binds to the endothelial lipase (EL) expressed on the surface of the Human Embryonic Kidney (HEK) 293 cells (the HEK 293 cell line has been engineered to express EL on the cell surface). The cell-bound MEDI5884 is then measured with a fixed concentration of an Eu-N1 anti-human IgG4P, which binds to the Fc region of the MEDI5884. The Eu-N1 anti-human IgG4P bound to MEDI5884 is measured by time-resolved fluorescence and the signal is directly proportional to MEDI5884 concentration.
MEDI5884 was evaluated through a pre-clinical developability study to understand the formulation development risks. The developability study was performed at 100 mg/ml in 20 mM His/His-HCl, 240 mM Sucrose, 0.02% (w/v) PS80, pH 6.0. Based on the developability study, MEDI5884 was found to be developable, however with a relatively high viscosity (12.6 cP at 23° C.) and Kd value (−19.6 mL/g). Based on this information and the anticipated clinical dose requirements (subcutaneous administration), a concentration of 100 mg/ml MEDI5884 following reconstitution of a lyophilized Drug Product (DP) was selected for use in a Phase 1 clinical trial.
The PS80 level in the drug substance (DS) was set to 0.02% (w/v), since this is known to provide adequate stability and protection from interfacial stress in most instances. However, the PS80 concentration in drug product (DP) post reconstitution is 0.04% (w/v) due to the half reconstitution approach.
Initial formulation development was performed as summarized in Table 7 below. However, the drug substance (DS) formulation #1 showed precipitation and cloudiness during the tangential flow filtration (TFF) process. In addition, the drug product (DP) formulation #1 showed presence of reversible self-association (RSA) by Dynamic Light Scattering (DLS;
Based on results obtained with DLS & AUC, the Arginine DP formulation (#3) showed absence of RSA (
A lyophilized formulation of MEDI5884, at a composition (post reconstitution) of 100 mg/mL in 20 mM Histidine/Histidine-HCl, 160 mM Arginine-HCl, 0.04% (w/v) Polysorbate, pH 6.0, provides appropriate stability and solubility and was suitable as a formulation for phase 1 clinical trial material supply.
MEDI5884 can be stored long term as Drug Substance (DS; formulated with Polysorbate 80) and Unformulated Drug Substance (UDS; formulated without Polysorbate 80) in a Celsius pak at 2-8° C. and at −40° C., respectively.
All the materials used for the study in this example were of USP or Multicompendial grade. All the solutions and buffers were prepared using USP or HPLC water and were filtered through 0.2 m PVDF filters (Millipore, Millex GV, SLG033RB) before further use. MEDI5884 samples for stability studies were prepared by following aseptic processing in a Biosafety Cabinet (BSC). Bulk material was stored at 2-8° C.
MEDI5884 protein concentrations were determined by measuring absorbance at 280 nm with a Trinean HT-A280 spectrophotometer (Gentbrugge, Belgium). A measured extinction coefficient of 1.44 (mg/mL)−1 cm−1 was used to calculate protein concentrations. As measurement of MEDI5884 on the Trinean HT-A280 does not require dilution, density correction factors were not used.
Size Exclusion Chromatography (SEC) was used for purity determination by high pressure size exclusion chromatography (HPSEC). An Agilent HPLC system (Santa Clara, Calif.) with a TSK-Gel G3000 column was used for analysis. 250 μg of antibody was loaded on the column by injecting 25 ul of the antibody diluted to 10 mg/ml in PBS. Standard integration parameters were used for automatic integration of aggregate, monomer and fragment peaks.
iii) Visual Appearance
Visual inspection of the samples was performed by examining the samples in their respective container for particles, color, and clarity using appropriate standards.
Sub-visible particles analysis was performed using light obscuration. For high accuracy (HIAC) analysis, samples were diluted to 5 mg/mL with formulation buffer and degassed for a minimum of 1 hour prior to testing.
Osmolality was measured using Advanced Instrument Inc. 2020 freezing point depression osmometer (Norwood, Mass.). System suitability was assessed by running a reference standard.
The viscosity of MEDI5884 Drug Product at 100 mg/mL was measured using an Anton Paar MCR301 Rheometer (Graz, Austria) with 40 mm cone and plate accessory. Viscosities are reported at the high-shear limit of 1000 per second shear rate.
vii) Lyophilization of Product Vials
Lyophilization was performed using a Lyostar III lyophilizer (SP Scientific, Warminster, Pa.).
viii) Determination of the Glass Transition Temperature (Tg′) for Lyophilization
Tg′ was determined using a TA Instruments Differential Scanning Calorimetry Instrument, Model Q2000 (New Castle, Del.). About 20 μL of sample was loaded in the pan. The samples were cooled to −80° C. at 5° C./min. They were then heated to 20° C. at 5° C./min to pass through the glass transition temperature. The Tg′ was determined as the sigmoidal curve/inflection in the heat flow curve. Integration for Tg′ was performed using Universal Analysis software to determine the onset, mid and end of glass transition and midpoint Tg′ values were reported as (Tg′).
The residual water present in the freeze-dried formulations was measured using Karl Fischer titration using a Mettler Toledo, model DL39 (Columbus, Ohio). Dry methanol was used to reconstitute the freeze-dried material. The residual water was determined by mass balance.
Freeze drying microscopy was performed using a liquid nitrogen vapor cooled Olympus Microscope (Tokyo, Japan) with sample holder capable of maintaining a vacuum on the sample. About 5-10 μL of sample was frozen to −40° C. and then the vacuum was started and the drying front of the cake was used to focus the microscope. Iterative ramp increases in temperature (fastest=2° C./min; slowest=0.5° C./min) were used to observe collapse in the microscope images of the drying cake. Onset of collapse (Tc) was reported as collapse temperature, which was observed as bright visible holes in the dried matrix at the sublimation front.
Stability of MEDI5884 lyophilized Drug Product was assessed at 100 mg/ml formulated in 20 mM His/His-HCl, 160 mM Arg-HCl, 0.04% (w/v) PS80, pH 6.0. Samples were placed in stability chambers at 40° C./75% relative humidity (RH), 25° C./60% RH and 5° C.
i) DS and UDS Storage and Strategy
DS and UDS storage in Celsius pak at 2-8 and −40° C. respectively were selected to assess long-term storage stability of MEDI5884. Controlled Freeze/Thaw assessment was performed for DS and UDS in the Celsius-paks container.
i) Controlled DS & UDS Freeze Thaw (F/T) and Frozen Storage Assessment in Celsius Pak
Freeze thaw and frozen storage stability were assessed in a representative storage container (i.e., Celsius pak), temperature and process. Celsius paks were filled with UDS and DS and frozen and thawed in a controlled manner three times. After the completion of third thaw, the Celsius paks were frozen again and stored at −40 and 25° C. for long term stability assessment for DS and UDS. For UDS, long term stability was also performed at 2-8 C and one month frozen storage data is shown below in Table 8.
DS showed no significant change in purity by size exclusion chromatography (SEC). Based on this assessment, tuED5884 is stable as a frozen DS in Celsius paks at −40° C. Also, there was no impact on stability for DS post 3×FT after 3 month of storage at 25° C.
Liquid Drug Substance stability is assessed in vials to support liquid hold times during storage, shipment and manufacturing. The liquid formulation stability was performed at 2-8, 25 and 40° C. in 3 cc glass vials with 1.0 mL fill volume. Results are shown in Table 9.
There was no significant change in pH, osmolality or protein concentration during the study, and visible and sub-visible particles remained low.
Viscosity measured at 23° C. was 5.4 cP for MEDI5884 at 100 mg/mL in 20 mM Histidine/Histidine-HCl, 160 mM Arginine-HCl, 0.04% (w/v) Polysorabte 80, pH 6.0. No challenges are expected in manufacturing or subcutaneous dose administration based on these viscosity measurements.
Drug product stability was assessed by SEC, visual inspection and melt flow index (MFI). Key results are reported below in Tables 10 and 11. These results support the use of this lyophilized drug product for Phase 1 clinical trials.
pH, osmolality and protein concentration remained unchanged during the DP stability study. Reconstitution time was in the range of 4 to 10 mins across the stability study and did not change significantly with time on stability. Moisture content for lyophilized cake was <1% for all samples in the study.
Post-reconstitution stability was assessed by SEC, visual inspection and MFI in order to support in-use stability in the clinic as well as hold times for analytical testing. Vials were reconstituted and tested at t0 and material was subsequently held in the vial at 5° C. and 25° C. for up to 1 week. Stability is maintained in both cases with no significant change in monomer purity and low visible and sub-visible particle levels comparable to t0. Data are shown in Table 12 below. Protein concentration, pH and osmolality were unchanged during the study (data not shown). Therefore, the material can be held at 5° C. up to 7 days without concerns regarding stability of reconstituted drug product.
This example describes the development of formulations of MEDI5884 having suitable viscosities for use in autoinjector devices for subcutaneous administration. Certain parameters of the formulation were tested for their effect on the viscosity of the formulation. All methods performed in this example were performed as described in Example 3 unless otherwise indicated.
a) pH Buffer Screen
This study was performed in order to determine a suitable pH for the product formulation. Formulations were tested for the viscosity effects of three different buffers each at three different pH values. Formulations tested contained either 100 mg/mL or 165 mg/mL MEDI5884 in 160 mM ArgHCl and 20 mm of the buffer indicated at the pH indicated. Tests were performed at 18° C. Results are shown in
b) Excipient Screen
Formulations were tested for the viscosity effects of various excipients. The formulations tested had a MEDI5884 concentration of 150 mg/mL in 20 mM His/His-HCl at pH 6.0. Tests were performed at 18° C. All excipients were present at a concentration of 190 mM except for DMSO, which was at a concentration of 5%. A Histidine only formulation was tested having no additional excipient added. Excipients tested were: the positively charged amino acids lysine HCl (Lys HCl) and arginine HCl (Arg HCl); the hydrophobic amino acid proline; the negatively charged amino acid glutamic acid (as sodium glutamate); the salts sodium sulfate (Na2SO4) and sodium chloride (NaCl); and the cosolvent dimethyl sulfoxide (DMSO). Results of the excipient screen are shown in
C) Determination of Viscosity at Different ArgHCl Concentrations
Formulations were tested for the viscosity effects of different concentrations of ArgHCl excipient in order to determine a suitable concentration of ArgHCl. The formulations tested had a MEDI5884 concentration of 145, 165 or 175 mg/mL in 20 mM His/HisHCl 0.02% PS80 and the concentrations of ArgHCl indicated. Formulations had a pH of 6.0 and were tested at 18° C. Results of this viscosity screening are shown in
d) pH Robustness Screen
Formulations were tested for the effects of variations in pH. The formulations tested had a MEDI5884 concentration of 165 mg/mL in 20 mM His/HisHCl and 220 mM ArgHCl in either His or acetate buffer at the pH values indicated. Tests were performed at 18° C. Results of the pH robustness screen are shown in
e) Determination of Viscosity at Different MEDI5884 Concentrations
Formulations having concentrations of greater than 100 mg/mL MEDI5884 may be needed for use in autoinjector delivery systems. This viscosity study was conducted in order to determine suitable concentrations of MEDI5884 in the formulation. The viscosity and glide force (GF) of formulations having varying concentrations of MEDI5884 were tested at concentrations of 135, 150 and 165 mg/mL MEDI5884 in 20 mM His/His-HCl, 220 mM Arg-HCl, 0.02% (w/v) PS80 at pH 6.0. Viscosity at 18° C. was measured using the methods described in Example 3. Glide force was also measured using pre-filled syringes (PFS) containing the formulation at 18° C. using Instron. Results are shown in Table 13.
f) Drug Product Stability Studies
The stability of drug product (DP) formulations was tested using the stability study methods described in Example 3. As the target formulation had a MED5884 concentration of 125 mg/mL, bracketing concentrations of 110 mg/mL and 140 mg/mL were used. Each formulation had a concentration of 20 mM Histidine/Histidine HCl, 220 mM Arg-HCl and 0.02% (w/v) PS80 at pH 6.0. Studies were performed at 5° C. for 16 months, 25° C. for 6 months and 40° C. for 3 months. The rate of monomer loss per month, percent aggregation per month and fragmentation per month were all measured. Results are shown in Table 14 below.
As can be seen in Table 14, no significant change in monomer purity was seen after 16 months at 2-8 C, and there were low aggregation and fragmentation rates at 25° C. and 40° C.
It will be readily apparent to one of ordinary skill in the relevant arts that other suitable modifications and adaptations to the methods and applications described herein can be made without departing from the scope of any of the embodiments. The above examples are included herewith for purposes of illustration only and are not intended to be limiting.
It is to be understood that while certain embodiments have been illustrated and described herein, the claims are not to be limited to the specific forms or arrangement of parts described and shown. In the specification, there have been disclosed illustrative embodiments and, although specific terms are employed, they are used in a generic and descriptive sense only and not for purposes of limitation. Modifications and variations of the embodiments are possible in light of the above teachings. It is therefore to be understood that the embodiments may be practiced otherwise than as specifically described.
While various embodiments have been described above, it should be understood that they have been presented only as illustrations and examples of the present technology, and not by way of limitation. It will be apparent to persons skilled in the relevant art that various changes in form and detail can be made therein without departing from the spirit and scope of the present technology. Thus, the breadth and scope of the present technology should not be limited by any of the above-described embodiments, but should be defined only in accordance with the appended claims and their equivalents. It will also be understood that each feature of each embodiment discussed herein, and of each reference cited herein, can be used in combination with the features of any other embodiment. All patents and publications discussed herein are incorporated by reference herein in their entirety.
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/US21/19213 | 2/23/2021 | WO |
| Number | Date | Country | |
|---|---|---|---|
| 62980658 | Feb 2020 | US |
