EA046788B1 - COMPOSITIONS OF CXCR4 INHIBITORS, METHODS OF PREPARATION AND APPLICATION - Google Patents

Abstract

The invention provides a compound of formula X4P-001 and pharmaceutical compositions containing the compound of formula I and said compound. The proposed compounds are characterized by reduced impurity levels, and compositions based on them are suitable for the treatment, prevention, or amelioration of a disease, disorder, or condition associated with a chemokine receptor, such as CXCR4.

Description

Field of technology

The present invention provides compounds that inhibit C-X-C chemokine receptor type 4 (CXCR4). The present invention also provides pharmaceutically acceptable compositions containing the compounds of the present invention and methods of using said compositions for the treatment of various disorders.

Cross reference to related applications

This application claims priority to U.S. Provisional Patent Application No. 62726010, filed Aug. 31, 2018, and also claims priority to U.S. Patent Application No. 16/215963, filed Dec. 11, 2018; the contents of which are incorporated herein by reference.

State of the art

C-X-C chemokine receptor type 4 (CXCR4), also known as fusin or cluster of differentiation 184 (CD184), is a seven-helix transmembrane G protein-coupled receptor (GPCR) belonging to class I GPCRs or the rhodopsin-like GPCR family. Under normal physiological conditions, CXCR4 performs several functions. functions and is mainly expressed in the hematopoietic and immune systems. CXCR4 was originally discovered as one of the coreceptors involved in human immunodeficiency virus (HIV) entry into cells. Subsequent studies have shown that this receptor is expressed in many tissues, including the brain, thymus, lymphatic tissues, spleen, stomach and small intestine, as well as certain cell types such as hematopoietic stem cells (HSC), mature lymphocytes and fibroblasts. CXCL12, previously designated SDF-1a, is the only known ligand for CXCR4. CXCR4 mediates stem cell migration during embryonic development, as well as during injury and inflammation. CXCR4 has been shown to play various roles in human diseases such as cell proliferative disorders, Alzheimer's disease, HIV, rheumatoid arthritis, pulmonary fibrosis, and others. For example, expression of CXCR4 and CXCL12 has been observed in several tumor types. CXCL12 is expressed by cancer-associated fibroblasts (CAFs) and is often present at high levels in the tumor microenvironment (TME). In clinical studies of a wide range of tumor types, including breast, ovarian, kidney, lung and melanoma, CXCR4/CXCL12 expression is associated with poor prognosis and an increased risk of metastases in lymph nodes, lung, liver and brain, which are the sites of expression of CXCL12. CXCR4 is frequently expressed in melanoma cells, especially in the CD133+ population, which is thought to represent melanoma stem cells. Experiments in vitro and in mouse models have demonstrated that CXCL12 is chemotactic for such cells.

In addition, there is now evidence implicating the CXCL12/CXCR4 axis in loss or failure of tumor response to angiogenesis inhibitors (also called angiogenic escape). In animal models of cancer, interference with CXCR4 function has been shown to alter the TME and sensitize tumors to immune attack through multiple mechanisms, such as eliminating tumor neovascularization and increasing the ratio of CD8+ T cells to Treg cells. These effects lead to significant reductions in tumor burden and increased overall survival in xenograft, syngeneic, and transgenic cancer models. See Vanharanta et al. (2013) Nat Med 19: 50-56; Gale and McColl (1999) BioEssays 21: 17-28; Highfill et al. (2014) Sci Transl Med 6:ra67; Facciabene et al. (2011) Nature 475: 226-230.

These data highlight the significant unmet need for CXCR4 inhibitors to treat many diseases and conditions mediated by aberrant or unwanted expression of the receptor, such as cell proliferative disorders.

Brief description of the invention

In the present invention, it has been found that the X4P-001 compositions of the present invention and pharmaceutically acceptable compositions thereof are effective as CXC receptor type 4 (CXCR4) inhibitors. In one aspect, the present invention provides composition X4P-001 containing a compound of formula I

I or a pharmaceutically acceptable salt thereof;

and at least one connection selected from the following:

- 1 046788

1-6 1-7 or a pharmaceutically acceptable salt thereof.

The X4P-001 compositions of the present invention and pharmaceutically acceptable compositions thereof are suitable for use in the treatment of various diseases, disorders or conditions associated with CXCR4, such as hyperproliferative conditions, including various cancers. Such diseases, disorders or conditions include the diseases, disorders or conditions described herein.

Brief description of drawings

In fig. 1 provides a brief description of the process for obtaining capsules containing the solid form of the medicinal product (single dosage form) X4P-001.

In fig. Figure 2 shows a schematic diagram of the original synthesis method (method 2 version or method 2) used to obtain X4P-001 for clinical studies.

In fig. 3 shows a comparison of method 3 (according to the invention, improved synthesis) and method 2 (original method) for obtaining X4P-001.

In fig. Figure 4 provides a detailed comparison of versions of method 2 and 3 for sequential top-down operations to isolate and extract APIs.

In fig. Table 5 shows HPLC and MS data for X4P-001, PTL/ST/0511, batch 3-1 (obtained using method 2), 25°C/60% RH, t=3 months. HPLC conditions 1 (detailed below); 1 mg/ml in methanol, injected sample volume 100 µl.

In fig. Table 6 shows HPLC and MS data for X4P-001, PTL/ST/0511, batch 3-1 (obtained using method 2), 25°C/60% RH, t=3 months. HPLC conditions 2 (detailed below); 1 mg/ml in methanol, injected sample volume 100 µl.

In fig. Table 7 shows HPLC and MS data for X4P-001, PTL/ST/0511, batch 3-1 (obtained using method 2), 25°C/60% RH, t=3 months. HPLC conditions 2 (detailed below); sample concentration in methanol 10 mg/ml, injected sample volume 100 µl.

In fig. 8 shows HPLC and MS data for X4P-001, sample after decomposition 80°C/RH 80%, t=1 day. HPLC conditions 2 (detailed below); sample concentration in methanol 10 mg/ml, injected sample volume 100 µl.

In fig. Figure 9 shows HPLC and MS data for X4P-001, sample after decomposition 80°C/RH 80%, t=7 days. HPLC conditions 2 (detailed below); sample concentration in methanol 10 mg/ml, injected sample volume 100 µl.

In fig. 10 shows HPLC and MS data for X4P-001, PTL/ST/0511, batch 3-1 (obtained using method 2), 25°C/60% RH, t=3 months. HPLC conditions 3 (detailed below); sample concentration in methanol 10 mg/ml, injected sample volume 100 µl.

Detailed Description of the Invention

1. General Description of Certain Aspects of the Present Invention In one aspect of the present invention there are provided compounds and compositions thereof suitable for use in treating, pre-

- 2 046788 preventing and/or reducing the risk of developing a disease, disorder or condition in the pathogenesis of which CXCR4 is involved. In some embodiments, such compounds include compounds of the formulas described in the present invention, or pharmaceutically acceptable salts thereof.

In another aspect, the present invention provides compositions, including formulations and unit dosage forms, containing X4P-001 (ie, the compound of Formula I, the structure of which is set forth below) or a pharmaceutically acceptable salt thereof, wherein such compositions exhibit an improved purity profile. In some embodiments, the proposed composition X4P-001 exhibits reduced levels of known impurities, such as those described in the present invention, and/or reduced levels of unknown impurities compared to similar compositions produced by conventional methods.

In another aspect of the present invention there is provided a composition X4P-001 containing a compound of formula I

or a pharmaceutically acceptable salt thereof;

and at least one compound selected from the following compounds:

or a pharmaceutically acceptable salt thereof.

2. Definitions

The compounds of the present invention include those generally described above and are further illustrated by the classes, subclasses and types described herein. For the purposes of this specification, the following definitions shall be used unless otherwise noted. For the purposes of the present invention, the chemical elements are defined according to the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, ^75th Ed. Additionally, general principles of organic chemistry are described in Organic Chemistry, Thomas Sorrell, University Science Books, Sausalito: 1999 and March's Advanced Organic Chemistry, ^5th Ed., Ed.: Smith, M.B. and March, J., John Wiley & Sons , New York: 2001, the entire contents of each of which are incorporated herein by reference. As used herein, the term composition X4P-001 or the proposed composition X4P-001 refers to a composition containing a compound of formula I (i.e., X4P-001) or a pharmaceutically acceptable salt thereof in combination with at least one additional compound selected from I-1, I-2, I-3, I-4, I-5, I-6 or I-7. For the avoidance of doubt, the pharmaceutical composition of the proposed composition X4P-001 refers to a composition containing a compound of formula I (i.e., X4P-001) or a pharmaceutically acceptable salt thereof in combination with at least one additional compound selected from I-1, I -2, I-3, I-4, I-5, I-6 or I-7, with

- 3 046788 together with a pharmaceutically acceptable excipient, for example an adjuvant, excipient, binder, carrier or transfer medium.

For the avoidance of doubt, and without being limited by theory, it is believed that compound I-1 at least partially exists in isomeric (eg, tautomeric) forms; for example, compound I-1 is believed to undergo the following interconversion to a compound of structure X-1a

1-1 ^Eeyore

Thus, it should be understood that the individual isomers, as well as tautomeric and other isomeric forms of I-1 and La, are included within the scope of the present invention. The proposed composition X4P-001 may contain additional known or unknown impurities. As used herein, the term impurity includes one or more degradation products that occur during storage of X4P-001, and/or one or more by-products generated in the chemical reaction used in the production of X4P-001. In some embodiments, the impurity results from oxidation initiated by decomposition light, reaction with a residual solvent such as water or isopropyl acetate, a side reaction that occurs during the process used to prepare X4P-001, or reaction of X4P-001 with an excipient, present in the pharmaceutical composition X4P-001.

As used herein, the term inhibitor is defined as a compound that binds to and/or inhibits CXCR4 with measurable affinity. In some embodiments, the inhibitor has an IC ₅₀ and/or binding constant of less than about 100 μM, less than about 50 μM, less than about 1 μM, less than about 500 nM, less than about 100 nM, less than about 10 nM, or less than about 1 nM.

As used herein, the term pharmaceutically acceptable salt refers to salts that, based on careful medical evaluation, are suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic reaction, or the like, and have an acceptable benefit/risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, S. M. Berge et al. pharmaceutically acceptable salts are described in detail in J. Pharmaceutical Sciences, 1977, 66, 1-19, incorporated herein by reference. Pharmaceutically acceptable salts of the compounds of the present invention include salts derived from suitable inorganic and organic acids and bases.

Examples of pharmaceutically acceptable, non-toxic acid addition salts include basic group (eg, amino) salts prepared using inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid, or using organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid, or by other methods known in the art, such as ion exchange. Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, besylate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentane propionate, digluconate, dodecyl sulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxyethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, mesylate, 2-naphthalene sulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate and the like.

Salts derived from suitable bases include alkali metal, alkaline earth metal, ammonium and N ⁺ (C _1-4 alkyl) ₄ salts. Typical alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium and the like. Other pharmaceutically acceptable salts, as appropriate, include non-toxic ammonium, quaternary ammonium and amine salts prepared using counterions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, (C _1-6 alkyl)sulfonate and arylsulfonate.

It is also intended that, unless otherwise indicated, the structures provided herein include all isomeric (e.g., enantiomeric, diastereomeric, and geometric (or conformational)) forms of said structures, e.g., R- and S-configurations for each center of asymmetry, Z- and E-isomers based on the position of the substituents relative to the double bond and conformational Z- and E-isomers. Thus, individual stereochemical isomers, as well as mixtures of enantiome

- 4 046788 diastereomers and geometric (or conformational) isomers of the compounds according to the present invention are included in the scope of the present invention. Unless otherwise indicated, all tautomeric forms of the compounds of the present invention are included within the scope of the present invention.

As used herein, a therapeutically effective amount or effective amount means an amount of a substance (eg, a therapeutic agent, composition and/or formulation) that produces a desired biological response. In some embodiments, a therapeutically effective amount of a substance is an amount that is sufficient when administered as part of a dosage regimen to a subject suffering from or susceptible to a disease, condition, or disorder to treat, diagnose, prevent, and/or delay the onset of said disease, conditions or disorders. As will be appreciated by those skilled in the art, the effective amount of a substance may vary depending on factors such as the desired biological outcome, the substance being delivered, the target cell or tissue, etc. For example, an effective amount of a compound in a formulation for treating a disease, condition or disorder is an amount that alleviates, mitigates, attenuates, suppresses, prevents, delays the onset of, reduces the severity and/or reduces the incidence of one or more symptoms or signs of said disease, condition or disorders. In some embodiments, a therapeutically effective amount is at least a minimum amount of a compound or composition containing a compound that is sufficient to treat one or more symptoms of a disease or disorder.

As used herein, the terms treating, treating, and administering treatment refer to partially or completely alleviating, suppressing, delaying the onset of, preventing, mitigating, and/or ameliorating a disease or disorder or one or more symptoms of a disease or disorder described herein. In some embodiments, treatment may be administered after the development of one or more symptoms. In some embodiments, the term providing treatment includes preventing or stopping the progression of a disease or disorder. In other embodiments, treatment may be provided in the absence of symptoms. For example, treatment may be provided to a susceptible individual prior to the onset of symptoms (eg, in light of a history of symptoms and/or in light of genetic factors or other susceptibility factors). Treatment may also be continued after symptoms have resolved, for example to prevent or delay their recurrence. Thus, in some embodiments, the term providing treatment includes preventing the recurrence or recurrence of a disease or disorder.

As used herein, the term CXCR4-mediated in relation to a disorder, disease and/or condition means any disease, disorder or condition in which CXCR4 or a mutant thereof is known to play a role. Accordingly, another embodiment of the present invention relates to the treatment or reduction of the severity of one or more diseases in which CXCR4 or a mutant thereof is known to play a role. CXCR4-mediated also includes diseases, disorders and conditions in which the CXCR4/CXCL12 axis is involved.

As used herein, the term unit dosage form refers to a physically discrete unit of therapeutic composition suitable for the subject being treated. However, it should be understood that the total daily intake of the X4P-001 compositions of the present invention will be determined by the attending physician based on a thorough medical evaluation. The specific effective dose for any particular subject or organism will depend on a number of factors, including the disorder being treated and the severity of said disorder; the activity of the particular active agent used; the specific composition used; age, body weight, general health, gender and diet of the subject; timing of administration and rate of elimination of the particular active agent used; duration of treatment; drugs and/or additional therapies used in combination or in parallel with the particular compound(s) employed, and other similar factors well known in the medical field.

3. Description of Illustrative Connections

The present invention has found that the compounds of the present invention and their compositions are suitable for use in the treatment, prevention and/or reduction of the risk of developing a disease, disorder or condition associated with CXCR4, or in which CXCR4 is associated with pathogenesis.

In one aspect of the present invention, there is provided a composition comprising X4P-001 or a pharmaceutically acceptable salt thereof with a purity of at least 98.5%. In some embodiments, X4P-001 in the specified composition has a purity of at least 98.5% and contains less than 1.5% wt./wt. impurities. In some embodiments, the composition comprises X4P-001 or a pharmaceutically acceptable salt thereof with a purity of at least 98.6%, 98.7%, 98.8%, 98.9%, 99.0%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8% or 99.9%.

In one aspect of the present invention there is provided a composition X4P-001 containing a compound of formula I

or a pharmaceutically acceptable salt thereof;

and a detectable amount of at least one of the following compounds:

or pharmaceutically acceptable salts thereof;

and wherein the composition X4P-001 does not contain the following compound in a detectable amount:

1-4 or a pharmaceutically acceptable salt thereof.

In some embodiments, the X4P-001 composition contains each of I-1, I-2, I-3, I-5, I-6, and I7 or a pharmaceutically acceptable salt thereof.

In some embodiments, the amount of I-1 or a pharmaceutically acceptable salt thereof is less than about 0.5% w/w. compositions Х4Р-001.

In some embodiments, the amount of I-2 or a pharmaceutically acceptable salt thereof is less than about 0.3% w/w. compositions Х4Р-001.

In some embodiments, the amount of I-3 or a pharmaceutically acceptable salt thereof is less than about 0.4% w/w. compositions Х4Р-001.

In some embodiments, the amount of I-5 or a pharmaceutically acceptable salt thereof is less than about 0.4% w/w. compositions Х4Р-001.

In some embodiments, the amount of I-6 or a pharmaceutically acceptable salt thereof is less than about 0.4% w/w. compositions Х4Р-001.

In some embodiments, the amount of I-7 or a pharmaceutically acceptable salt thereof is less than about 0.25% w/w. compositions Х4Р-001.

In some embodiments, the amount of I-1 or a pharmaceutically acceptable salt thereof is from about 0.02 to about 0.5% w/w. compositions Х4Р-001.

- 6 046788

In some embodiments, the amount of I-2 or a pharmaceutically acceptable salt thereof is from about 0.01 to about 0.3% w/w. compositions Х4Р-001.

In some embodiments, the amount of I-3 or a pharmaceutically acceptable salt thereof is from about 0.01 to about 0.4% w/w. compositions Х4Р-001.

In some embodiments, the amount of I-5 or a pharmaceutically acceptable salt thereof is from about 0.01 to about 0.4% w/w. compositions Х4Р-001.

In some embodiments, the amount of I-6 or a pharmaceutically acceptable salt thereof is from about 0.01 to about 0.4% w/w. compositions Х4Р-001.

In some embodiments, the amount of I-7 or a pharmaceutically acceptable salt thereof is from about 0.01 to about 0.25% w/w. compositions Х4Р-001.

In one aspect of the present invention, there is provided a pharmaceutical composition comprising the disclosed composition X4P-001 and a pharmaceutically acceptable adjuvant, carrier or vehicle.

In some embodiments, the pharmaceutically acceptable adjuvant includes at least one diluent, disintegrant, lubricant, and flow agent.

In another aspect of the present invention, there is provided a composition X4P-001 or a pharmaceutical composition thereof, wherein:

(a) the amount of I-1 or a pharmaceutically acceptable salt thereof is less than about 0.5% w/w. compositions X4P-001;

(b) the amount of I-2 or a pharmaceutically acceptable salt thereof is less than about 0.3% w/w. compositions X4P-001;

(c) the amount of I-3 or a pharmaceutically acceptable salt thereof is less than about 0.4% w/w. compositions X4P-001;

(d) the amount of I-5 or a pharmaceutically acceptable salt thereof is less than about 0.4% w/w. compositions X4P-001;

(e) wherein the amount of I-6 or a pharmaceutically acceptable salt thereof is less than about 0.4% w/w. composition X4P-001 and (f) the amount of I-7 or a pharmaceutically acceptable salt thereof is less than about 0.25% w/w. compositions Х4Р-001.

In one aspect of the present invention, there is provided a composition of X4P-001 or a pharmaceutical composition thereof, wherein:

(a) the amount of I-1 or a pharmaceutically acceptable salt thereof is from about 0.02 to about 0.5% w/w. compositions X4P-001;

(b) the amount of I-2 or a pharmaceutically acceptable salt thereof is from about 0.01 to about 0.3% w/w. compositions X4P-001;

(c) the amount of I-3 or a pharmaceutically acceptable salt thereof is from about 0.01 to about 0.4% w/w. compositions X4P-001;

(d) the amount of I-5 or a pharmaceutically acceptable salt thereof is from about 0.01 to about 0.4% w/w, composition X4P-001;

(e) the amount of I-6 or a pharmaceutically acceptable salt thereof is from about 0.01 to about 0.4% w/w. composition X4P-001 and (f) the amount of I-7 or a pharmaceutically acceptable salt thereof is from about 0.01 to about 0.25% w/w. compositions Х4Р-001.

In one aspect of the present invention, composition X4P-001 is provided, wherein:

(a) the amount of I-2 or a pharmaceutically acceptable salt thereof is from about 0.01 to about 0.2% w/w. compositions X4P-001;

(b) the amount of I-3 or a pharmaceutically acceptable salt thereof is from about 0.01 to about 0.2% w/w. compositions X4P-001;

(c) the amount of I-5 or a pharmaceutically acceptable salt thereof is from about 0.01 to about 0.2% w/w. compositions X4P-001;

(d) the amount of I-6 or a pharmaceutically acceptable salt thereof is from about 0.01 to about 0.4% w/w. composition X4P-001 and (e) the amount of I-7 or a pharmaceutically acceptable salt thereof is from about 0.01 to about 0.25% w/w. compositions Х4Р-001.

In some embodiments, the X4P-001 composition optionally further contains I1 or a pharmaceutically acceptable salt thereof in an amount of from about 0.02 to about 0.5% w/w. compositions Х4Р-001. In another aspect of the present invention, there is provided a dosage unit form containing a pharmaceutical composition comprising:

(a) the proposed composition X4P-001 in an amount of approximately 10-20% by weight of the composition;

(b) microcrystalline cellulose in an amount of about 70-85% by weight of the composition;

(c) croscarmellose sodium in an amount of about 5-10% by weight of the composition;

(d) sodium stearyl fumarate in an amount of about 0.5-2% by weight of the composition and

- 7 046788 (e) colloidal silicon dioxide in an amount of approximately 0.1-1.0% by weight of the composition.

In another aspect of the present invention, there is provided a dosage unit form containing a pharmaceutical composition comprising:

(a) the proposed composition X4P-001 in an amount of approximately 35-75% by weight of the composition;

(b) microcrystalline cellulose in an amount of about 5-28% by weight of the composition;

(c) calcium hydrogen phosphate dihydrate in an amount of about 7-30% by weight of the composition;

(d) croscarmellose sodium in an amount of about 2-10% by weight of the composition;

(e) sodium stearyl fumarate in an amount of about 0.3-2.5% by weight of the composition;

(f) colloidal silica in an amount of about 0.05-1.2% by weight of the composition; and (g) sodium lauryl sulfate in an amount of about 0.2-1.2% by weight of the composition.

In one aspect, the present invention provides a method of treating, preventing, or reducing the risk of developing a disease, disorder, or condition associated with CXCR4 in a subject in need thereof, comprising administering to said subject an effective amount of the disclosed composition X4P-001.

In some embodiments, said disease, disorder, or condition is a cancer selected from kidney cancer, renal tumor, renal carcinoma, ovarian cancer, or melanoma.

In one aspect, the present invention provides composition X4P-001 containing a compound of formula I

or a pharmaceutically acceptable salt thereof;

and at least one connection selected from the following:

or a pharmaceutically acceptable salt thereof.

In another aspect of the present invention there is provided an X4P-001 composition containing a compound of formula I

- 8 046788

or a pharmaceutically acceptable salt thereof;

and at least one connection selected from the following:

or a pharmaceutically acceptable salt thereof.

In some embodiments, the present invention provides composition X4P-001 containing a compound of formula I

or a pharmaceutically acceptable salt thereof; and connecting the following structure:

or a pharmaceutically acceptable salt thereof. In some embodiments, the total weight of I-6 and any additional impurities present is no more than about 0.8% w/w of composition X4P-001.

In some embodiments, the X4P-001 composition contains I-6 or a pharmaceutically acceptable salt thereof in at least a detectable amount.

In some embodiments, the X4P-001 composition contains two, three, or four compounds selected from I-1, I-2, I-3, I-4, I-5, I-6, or I-7, or pharmaceutically acceptable ones thereof salts

In some embodiments, the X4P-001 composition contains at least a detectable amount of two, three, or four compounds selected from I-1, I-2, I-3, I-4, I-5, I-6, or I -7; or pharmaceutically acceptable salts thereof.

In some embodiments, the X4P-001 composition contains three compounds selected from I

- 9 046788

1, I-2, I-3, I-4, I-5, I-6 or I-7, or pharmaceutically acceptable salts thereof. In some embodiments, the X4P-001 composition contains at least a detectable amount of three compounds selected from I-1, I-2, I-3, I-4, I-5, I-6, or I-7, or pharmaceutical grade thereof acceptable salts.

In some embodiments, the X4P-001 composition contains each of I-1, I-2, I-3, I-5, I-6, and I7 or a pharmaceutically acceptable salt thereof. In some embodiments, the X4P-001 composition contains at least a detectable amount of each of I-1, I-2, I-3, I-5, I-6, and I-7, or a pharmaceutically acceptable salt thereof.

In one aspect, the present invention provides composition X4P-001 containing a compound of formula I

or a pharmaceutically acceptable salt thereof;

and a detectable amount of at least one of the following compounds:

or pharmaceutically acceptable salts thereof.

In some embodiments, the X4P-001 composition does not contain the following compound in a detectable amount:

or a pharmaceutically acceptable salt thereof.

In some embodiments, the X4P-001 composition does not contain the following compound in a detectable amount:

- 10 046788

or a pharmaceutically acceptable salt thereof.

In some embodiments, the total amount of I-6 or a pharmaceutically acceptable salt thereof is no more than about 0.2% w/w. composition X4P-001 relative to the total weight in composition X4P-001 of the compound of formula I and one or more compounds selected from I-1, I-2, I-3, I-4, I-5, I-6 or I-7 ; or pharmaceutically acceptable salts thereof. In some embodiments, the X4P-001 composition contains at least a detectable amount of I-6 or a pharmaceutically acceptable salt thereof. In some embodiments, the X4P-001 composition contains no more than 0.15% wt./wt. I-6 relative to the total weight in composition X4P-001 of the compound of formula I and one or more compounds selected from I-1, I-2, I-3, I-4, I-5, I-6 or I-7; or pharmaceutically acceptable salts thereof. In some embodiments, the X4P-001 composition contains at least a detectable amount of I-6 or a pharmaceutically acceptable salt thereof. In some embodiments, the total amount of I-6 or a pharmaceutically acceptable salt thereof is no more than about 0.2% w/w. composition X4P-001 relative to the total weight in composition X4P-001 of the compound of formula I and one or more compounds selected from I-2, I-3, I-5, I-6 or I-7; or pharmaceutically acceptable salts thereof. In some embodiments, the X4P-001 composition contains at least a detectable amount of I-6 or a pharmaceutically acceptable salt thereof. In some embodiments, the X4P-001 composition contains no more than 0.15% wt./wt. I-6 relative to the total weight in composition X4P-001 of the compound of formula I and one or more compounds selected from I-2, I-3, I-5, I-6 and I-7; or pharmaceutically acceptable salts thereof. In some embodiments, the X4P-001 composition contains at least a detectable amount of I-6 or a pharmaceutically acceptable salt thereof.

In some embodiments, one or more compounds selected from I-1, I-2, I-3, I-4, I-5, I6, or I-7; or their pharmaceutically acceptable salts together represent less than about 3.0% wt./wt. composition X4P-001 relative to the total weight in composition X4P-001 of the compound of formula I and one or more compounds selected from I-1, I-2, I-3, I-4, I-5, I-6 or I-7 ; or pharmaceutically acceptable salts thereof. In some embodiments, one or more compounds selected from I-1, I-2, 1I-3, I-4, I-5, I-6, or I-7; or pharmaceutically acceptable salts thereof are present in the composition X4P-001 in at least a detectable amount. In some embodiments, the X4P-001 composition contains a compound of Formula I or a pharmaceutically acceptable salt thereof and does not contain a compound of Formula I-4 or a pharmaceutically acceptable salt thereof in a detectable amount. In some embodiments, the total amount of organic impurities, including one or more compounds selected from I-1, I-2, I-3, I-4, I-5, I-6, or I-7; or pharmaceutically acceptable salts thereof, constitutes less than about 4.0% w/w of composition X4P-001.

In some embodiments, one or more compounds selected from I-2, I-3, I-5, I-6, and I-7; or their pharmaceutically acceptable salts together constitute less than about 3.0% wt./wt. composition X4P-001 relative to the total weight in composition X4P-001 of the compound of formula I and one or more compounds selected from I-2, I-3, I-5, I-6 or I-7; or pharmaceutically acceptable salts thereof. In some embodiments, one or more compounds selected from I-2, I-3, I-5, I-6, and I-7; or pharmaceutically acceptable salts thereof are present in the composition X4P-001 in at least a detectable amount. In some embodiments, the X4P-001 composition contains a compound of Formula I or a pharmaceutically acceptable salt thereof and does not contain either Compound I-1, Compound I-4, or their pharmaceutically acceptable salts in a detectable amount.

In some embodiments, the total amount of organic impurities including one or more compounds selected from I-2, I-3, I-5, I-6, or I-7; or their pharmaceutically acceptable salts, is less than about 4.0% wt./wt. compositions Х4Р-001.

In some embodiments, the amount of I-1 or a pharmaceutically acceptable salt thereof is less than about 0.5% w/w. compositions Х4Р-001.

In some embodiments, the amount of I-2 or a pharmaceutically acceptable salt thereof is less than about 0.3% w/w. compositions Х4Р-001.

In some embodiments, the amount of I-3 or a pharmaceutically acceptable salt thereof is less than about 0.4% w/w. compositions Х4Р-001.

In some embodiments, the X4P-001 composition does not contain I-4 or a pharmaceutically acceptable salt thereof in a detectable amount.

In some embodiments, the amount of I-5 or a pharmaceutically acceptable salt thereof with

- 11 046788 is less than about 0.07% w/w. compositions Х4Р-001.

In some embodiments, the amount of I-6 or a pharmaceutically acceptable salt thereof is less than about 0.4% w/w. compositions Х4Р-001.

In some embodiments, the amount of I-7 or a pharmaceutically acceptable salt thereof is less than about 0.25% w/w. compositions Х4Р-001.

In some embodiments, at least a detectable amount of I-1, I-2, I-3, I-5, I-6, and I-7 or pharmaceutically acceptable salts thereof is present in the X4P-001 composition.

In some embodiments, the amount of I-1 or a pharmaceutically acceptable salt thereof is from about 0.02 to about 0.5% w/w. compositions Х4Р-001.

In some embodiments, the amount of I-2 or a pharmaceutically acceptable salt thereof is from about 0.01 to about 0.3% w/w. compositions Х4Р-001.

In some embodiments, the amount of I-3 or a pharmaceutically acceptable salt thereof is from about 0.01 to about 0.4% w/w. compositions Х4Р-001.

In some embodiments, the amount of I-5 or a pharmaceutically acceptable salt thereof is from about 0.01 to about 0.07% w/w. compositions Х4Р-001.

In some embodiments, the amount of I-6 or a pharmaceutically acceptable salt thereof is from about 0.01 to about 0.4% w/w. compositions Х4Р-001.

In some embodiments, the amount of I-7 or a pharmaceutically acceptable salt thereof is from about 0.01 to about 0.25% w/w. compositions Х4Р-001.

In some embodiments, composition X4P-001 contains a compound of formula I or a pharmaceutically acceptable salt thereof with a purity of at least 99.3% by HPLC and contains less than 0.7% in total of additional compounds selected from I-1, I- 2, I-3, I-5, I-6 and I-7; or pharmaceutically acceptable salts thereof, as measured by HPLC. In some embodiments, at least a detectable amount of one or more of the additional compounds is present.

In some embodiments, composition X4P-001 contains a compound of formula I or a pharmaceutically acceptable salt thereof with a purity of at least 99.3% by HPLC and contains less than 0.7% in total of additional compounds selected from I-2, I- 3, I-5, I-6 or I-7; or pharmaceutically acceptable salts thereof, as measured by HPLC. In some embodiments, at least a detectable amount of one or more of the additional compounds is present. In some embodiments, at least a detectable amount of each of I-2, I-3, I-5, I-6, or I-7 or pharmaceutically acceptable salts thereof is present, as measured by HPLC.

In one aspect of the present invention there is provided a composition X4P-001 containing a compound of formula I

or a pharmaceutically acceptable salt thereof and one or more compounds selected from the following compounds:

or a pharmaceutically acceptable salt thereof in an amount of not more than about 0.5% w/w. compositions X4P-001;

- 12 046788

1-2 or a pharmaceutically acceptable salt thereof in an amount of not more than about 0.3% wt./wt. compositions X4P-001;

N/N X-NH

1-3 or a pharmaceutically acceptable salt thereof in an amount of not more than about 0.4% wt./wt. compositions X4P-001;

N ^z NH

1-5 or a pharmaceutically acceptable salt thereof in an amount of not more than about 0.5% w/w. compositions X4P-001;

1-6 or a pharmaceutically acceptable salt thereof in an amount of not more than about 0.4% wt./wt. composition X4P-001 or nh ₂ or a pharmaceutically acceptable salt thereof in an amount of not more than about 0.25% wt./wt. compositions X4P-001; where each % wt./wt. measured relative to the total weight in composition X4P-001 of the compound of formula I and one or more compounds selected from I-1, I-2, I-3, I-5, I-6 and I-7. In some embodiments, the X4P-001 composition contains a compound of Formula I or a pharmaceutically acceptable salt thereof and does not contain a compound of Formula I-4 or a pharmaceutically acceptable salt thereof in a detectable amount.

In one aspect of the present invention there is provided a composition X4P-001 containing a compound of formula I

- 13 046788

or a pharmaceutically acceptable salt thereof;

and one or more compounds selected from the following compounds:

or a pharmaceutically acceptable salt thereof in an amount of not more than about 0.3% w/w. compositions X4P-001;

1-3 or a pharmaceutically acceptable salt thereof in an amount of not more than about 0.4% wt./wt. compositions X4P-001;

1-5 or a pharmaceutically acceptable salt thereof in an amount of not more than about 0.5% w/w. compositions X4P-001;

1-6 or a pharmaceutically acceptable salt thereof in an amount of not more than about 0.4% wt./wt. compositions X4P-001; or

1-7 or a pharmaceutically acceptable salt thereof in an amount of not more than about 0.25% w/w of composition X4P-001; where each % wt./wt. measured relative to the total weight in the composition of the compound of formula I and one or more compounds selected from I-2, I-3, I-5, I-6 or I7. In some embodiments, the chiral purity of the X4P-001 composition is at least about 97% enantiomeric excess (% ee).

In some embodiments, the X4P-001 composition contains 7000, 6000, 5000, 4500, 4000, 3500, 3000, 2500, 2000, 1750, 1700, 1650, 1600, 1550, 1500, 1450, 1400, or ppm or less of toluene.

In some embodiments, toluene is used as a crystallization solvent to isolate X4P-001. In some embodiments, the specification for residual toluene in X4P-001 free base is such that the X4P-001 composition contains no more than 4500 ppm. In other embodiments, the X4P-001 composition contains no more than 4000 ppm, 3500 ppm, 3000 ppm, 2500 ppm, 2000 ppm, 1750 ppm, 1700 ppm, 1650 ppm, 1600 ppm, 1550 ppm, 1500 ppm, 1450 ppm, 1400 ppm or 1350 ppm toluene. Some implementations use a permissible daily exposure (PDE) approach. The term tolerable daily exposure (PDE) is defined as the pharmaceutically acceptable intake of residual solvents in a drug product. See, for example, Guidance for Industry: Q3C Impurities: Residual Solvents, published by the Food and Drug Administration (FDA) of the US Department of Health and Human Services.

In some embodiments, the % purity of the X4P-001 composition, as measured by HPLC, decreases by less than 1% when the X4P-001 composition is stored for three months at 25°C/60% relative humidity.

In some embodiments, the X4P-001 composition further contains one or more of the following compounds:

and wherein compounds I-8, I-9, I-10 and/or I-11 are present in an amount of less than about 25 parts per million (ppm) of the composition X4P-001.

In some embodiments, compounds I-8, I-9, I-10, and/or I-11 are present in an amount of less than about 50, 40, 30, 20, 15, 10, 5, 4, 3, 2, or 1 ppm compositions Х4Р-001. In some embodiments, compounds I-8, I-9, I-10, and/or I-11 are each independently present in an amount of from about 1 ppm to about 25 ppm, or from about 100 parts per billion (ppb) to 4 ppm. In some embodiments, the X4P-001 composition contains a compound of Formula I or a pharmaceutically acceptable salt thereof and does not contain a compound of Formula I-4 or a pharmaceutically acceptable salt thereof in a detectable amount.

In one aspect of the present invention there is provided a pharmaceutical composition comprising the proposed composition X4P-001 and a pharmaceutically acceptable adjuvant, carrier or carrier medium.

In one aspect of the present invention there is provided a solid unit dosage form prepared for oral administration containing the proposed composition X4P-001 or a pharmaceutical composition.

In some embodiments, the present invention provides the proposed composition of X4P-001 in combination with an additional therapeutic agent.

In another aspect of the present invention, there is provided a dosage unit form containing a pharmaceutical composition comprising:

(a) the proposed composition X4P-001 in an amount of approximately 10-20% by weight of the composition;

(b) microcrystalline cellulose in an amount of about 70-85% by weight of the composition;

(c) croscarmellose sodium in an amount of about 5-10% by weight of the composition;

(d) sodium stearyl fumarate in an amount of about 0.5-2% by weight of the composition; and (e) colloidal silicon dioxide in an amount of about 0.1-1.0% by weight of the composition.

In another aspect of the present invention, there is provided a dosage unit form containing a pharmaceutical composition comprising:

(a) the proposed composition X4P-001 in an amount of approximately 30-40% by weight of the composition;

(b) microcrystalline cellulose in an amount of about 20-25% by weight of the composition;

(c) calcium hydrogen phosphate dihydrate in an amount of about 30-35% by weight of the composition;

(d) croscarmellose sodium in an amount of about 5-10% by weight of the composition;

- 15 046788 (e) sodium stearyl fumarate in an amount of approximately 0.5-2% by weight of the composition;

(f) colloidal silica in an amount of about 0.1-1.0% by weight of the composition; and (g) sodium lauryl sulfate in an amount of about 0.1-1.0% by weight of the composition.

In another aspect of the present invention, there is provided a dosage unit form containing a pharmaceutical composition comprising:

(a) the proposed composition X4P-001 in an amount of approximately 35-75% by weight of the composition;

(b) microcrystalline cellulose in an amount of about 5-28% by weight of the composition;

(c) calcium hydrogen phosphate dihydrate in an amount of about 7-30% by weight of the composition;

(d) croscarmellose sodium in an amount of about 2-10% by weight of the composition;

(e) sodium stearyl fumarate in an amount of about 0.3-2.5% by weight of the composition;

(f) colloidal silica in an amount of about 0.05-1.2% by weight of the composition; and (g) sodium lauryl sulfate in an amount of about 0.2-1.2% by weight of the composition.

In some embodiments, said unit dosage form is in the form of a capsule.

In some embodiments, the capsule contains about 25 mg, about 100 mg, or about 200 mg of X4P-001 or a pharmaceutically acceptable salt thereof.

In another aspect, the present invention provides a method of treating, preventing, or reducing the risk of developing a disease, disorder, or condition associated with CXCR4 in a subject in need thereof, comprising administering to said subject the disclosed composition X4P-001.

In some embodiments, said disease, disorder, or condition is cancer.

In some embodiments, the cancer is selected from kidney cancer, renal tumor, renal carcinoma (including clear cell and papillary renal carcinoma), ovarian cancer, or melanoma.

In some embodiments, the X4P-001 composition comprises a compound of Formula I or a pharmaceutically acceptable salt thereof and one additional compound selected from I-1, I-2, I-3, I-4, I5, I-6, or I-7; or a pharmaceutically acceptable salt thereof.

In some embodiments, the X4P-001 composition contains a compound of Formula I or a pharmaceutically acceptable salt thereof and two additional compounds selected from I-1, I-2, I-3, I-4, I-5, I-6, or I- 7; or pharmaceutically acceptable salts thereof.

In some embodiments, the X4P-001 composition contains a compound of formula I or a pharmaceutically acceptable salt thereof and three additional compounds selected from I-1, I-2, I-3, I-4, I-5, I-6, or I- 7; or pharmaceutically acceptable salts thereof.

In some embodiments, composition X4P-001 contains a compound of formula I or a pharmaceutically acceptable salt thereof and four additional compounds selected from I-1, I-2, I-3, I-4, I-5, I-6, or I- 7; or pharmaceutically acceptable salts thereof.

In some embodiments, the X4P-001 composition contains a compound of Formula I or a pharmaceutically acceptable salt thereof and does not contain a compound of Formula I-4 or a pharmaceutically acceptable salt thereof in a detectable amount.

In some embodiments, said additional compound or compounds are present in at least a detectable amount.

In some embodiments, the composition of X4P-001 contains a compound of Formula I or a pharmaceutically acceptable salt thereof and each of I-1, I-2, I-3, I-4, I-5, I-6 and I-7 or a pharmaceutically acceptable salt thereof. acceptable salts.

In some embodiments, the X4P-001 composition comprises a compound of Formula I or a pharmaceutically acceptable salt thereof and one additional compound selected from I-2, I-3, I-5, I-6, or I-7; or a pharmaceutically acceptable salt thereof.

In some embodiments, the X4P-001 composition comprises a compound of Formula I or a pharmaceutically acceptable salt thereof and two additional compounds selected from I-2, I-3, I-5, I-6 and I7; or pharmaceutically acceptable salts thereof.

In some embodiments, the X4P-001 composition comprises a compound of formula I or a pharmaceutically acceptable salt thereof and three additional compounds selected from I-2, I-3, I-5, I-6 and I7; or pharmaceutically acceptable salts thereof.

In some embodiments, the X4P-001 composition contains a compound of formula I or a pharmaceutically acceptable salt thereof and four additional compounds selected from I-2, I-3, I-5, I-6 and I-7; or pharmaceutically acceptable salts thereof.

In some embodiments, the X4P-001 composition contains a compound of Formula I or a pharmaceutically acceptable salt thereof and does not contain a detectable amount of a compound of Formulas I-1 and I-4 or a pharmaceutically acceptable salt thereof.

In some embodiments, said additional compound or compounds are present in at least a detectable amount.

In some embodiments, the composition of X4P-001 contains a compound of formula I or a pharmaceutically acceptable salt thereof and each of I-2, I-3, I-5, I-6 and I-7 or a pharmaceutically acceptable salt thereof

- 16 046788 salts.

In some embodiments, the X4P-001 composition comprises a compound of Formula I or a pharmaceutically acceptable salt thereof and I-1 or a pharmaceutically acceptable salt thereof.

In some embodiments, the X4P-001 composition comprises a compound of Formula I or a pharmaceutically acceptable salt thereof and I-2 or a pharmaceutically acceptable salt thereof.

In some embodiments, the X4P-001 composition contains a compound of Formula I or a pharmaceutically acceptable salt thereof and I-3 or a pharmaceutically acceptable salt thereof.

In some embodiments, the X4P-001 composition contains a compound of Formula I or a pharmaceutically acceptable salt thereof and I-4 or a pharmaceutically acceptable salt thereof.

In some embodiments, the X4P-001 composition contains a compound of Formula I or a pharmaceutically acceptable salt thereof and I-5 or a pharmaceutically acceptable salt thereof.

In some embodiments, the X4P-001 composition contains a compound of Formula I or a pharmaceutically acceptable salt thereof and I-6 or a pharmaceutically acceptable salt thereof.

In some embodiments, the X4P-001 composition contains a compound of Formula I or a pharmaceutically acceptable salt thereof and I-7 or a pharmaceutically acceptable salt thereof.

In some embodiments, the composition of X4P-001 contains a compound of formula I or a pharmaceutically acceptable salt thereof and I-1, I-2, I-3, I-4, I-5, I-6 and I-7 or a pharmaceutically acceptable salt thereof .

In some embodiments, the % w/w amount of a compound in a subject composition X4P-001 is measured by comparing the area percentage of the compound on an HPLC chromatogram with any other compounds present in the composition X4P-001.

For example, when measured in this manner, 0.2% wt./wt. compound I-6 present in a composition X4P-001 containing I-6 and a compound of formula I means that said composition contains I-6 with a % peak area of 0.2% and a compound of formula I with a % peak area according to HPLC 99.8%. In other embodiments, % wt./wt. measured using other means known to one skilled in the art, such as those described in the present invention.

In one aspect of the present invention there is provided a method for producing the proposed composition X4P-001, wherein said composition is prepared essentially as described in the examples and figures herein.

In another aspect of the present invention, a compound selected from the compounds presented in table is provided. 1 below.

Table 1. Representative compounds of the present invention (5)-R-((177-benzo[<7]imidazol-2yl)methyl)-A'-(4(methyleneamino)butyl)-5,6,7,8tetrahydroquinoline-8 -amine 1-1 (5)-R-(4-(((1R-benzo[<7]imidazol-2yl)methyl)(5,6,7,8-tetrahydroquin olin-8yl)amino)butyl)formamide

1-2

-((1R-benzo[<7]imidazol-2yl)methyl)- 1R-benzo[<7]imidazol-2yl)methyl)-A ¹ -(5,6,7,8tetrahydroquin olin-8-yl)butan- 1,4diamine

1-3 (6)-4-(((4-(((1R-benzo[<7]imidazol-2yl)methyl)(5,6,7,8-tetrahydroquin olin-8yl)amino)butyl)amino) methoxy)benzoic acid (5)-R-(4-(((1R-benzo[<7]imidazol-2yl)methyl)(5,6,7,8(6)-4-(((1H-benzo[ c1]imidazol-2yl)methyl)(5,6,7,8-tetrahydroquine oline-8

- 17 046788 tetrahydroquinolin-8yl)amino)butyl)acetamide 1-5 yl)amino)butanal 1-6

(5)-M ¹ -(5,6,7,8-tetrahydroquinolin-8yl)butane-1,4-diamine 1-7

In some embodiments, the present invention provides the compound presented in table. 1 above, or a pharmaceutically acceptable salt thereof.

In some embodiments, the present invention provides a pharmaceutical composition that contains a compound of formula I or a pharmaceutically acceptable salt thereof and at least one of the compounds presented in table. 1 above, or pharmaceutically acceptable salts thereof. Said composition may contain 1, 2, 3, 4, 5, 6 or 7 of these compounds. In some embodiments, the composition does not contain Compound I-4 in a detectable amount. In some embodiments, the composition does not contain Compound I-1 in a detectable amount. In some embodiments, the composition does not contain compounds I-1 and I-4 in a detectable amount.

Typically, the wt.% of each impurity is determined by HPLC and measured either initially or after storage and optionally on an ongoing basis during the shelf life of the X4P001 composition. In some embodiments, the impurity content is measured after storing the composition under stress conditions, which are conditions of elevated temperature, humidity, or both, used to approximate the effect of long-term storage under ambient conditions. In some embodiments, the compound of Formula I, or a pharmaceutically acceptable salt thereof, is present in composition X4P-001 in an amount of at least about 96, 97, 97.5, 98, 98.5, 98.7, 98.9, 99.0, 99.1, 99.2, 99.3, 99.4, 99.5, 99.6, 99.7, 99.75, 99.8, 99.85, 99.9, 99.95, 99, 97 or 99.999 percent by weight, where the percentages are based on the free base of the compound and the total weight of the X4P-001 composition. In other embodiments, the X4P-001 composition contains no more than about 2.0 HPLC area percent total organic impurities, or in other embodiments no more than about 5.0, 4.0, 2.0, 1.9, 1.8, 1.7, 1.6, 1.5, 1.25, 1, 0.75, 0.5, 0.25, 0.2, 0.1, 0.01, 0.005 or 0.001 percent areas according to HPLC total organic impurities relative to the total area of the HPLC chromatogram.

In other embodiments, the X4P-001 composition contains no more than about 5.0, 4.0, 2.0, 1.9, 1.8, 1.7, 1.6, 1.5, 1.25, 1 , 0.75, 0.5, 0.25, 0.2, 0.1, 0.01, 0.005, or 0.001 area percent (measured by HPLC) of compounds I-1, I-2, I-3, I -4, I-5, I-6 and I-7 relative to the total area of the HPLC chromatogram. In other embodiments, the composition of X4P-001 contains a compound of Formula I or a pharmaceutically acceptable salt thereof and one or more additional compounds selected from I1, I-2, I-3, I-4, I-5, I-6, or I- 7; or pharmaceutically acceptable salts thereof and at least one pharmaceutically acceptable carrier. In some embodiments, the X4P-001 composition contains a compound of Formula I or a pharmaceutically acceptable salt thereof in an amount of from about 1 to about 99% by weight, where the percentages are based on the free base of the compound and the total weight of the X4P-001 composition. In other embodiments, the X4P001 composition contains no more than about 2.0 HPLC area percent total organic impurities, or in other embodiments no more than about 5.0, 4.0, 2.0, 1.9, 1. 8, 1.7, 1.6, 1.5, 1.25, 1.1, 1.05, 1, 0.95, 0.9, 0.8, 0.75, 0.7, 0, 6, 0.5, 0.25, 0.2, 0.1, 0.01, 0.005, or 0.001 HPLC area percent total organic impurities relative to the total area of the HPLC chromatogram. In some embodiments, one or more compounds selected from I-1, I-2, I3, I-4, I-5, I-6, or I-7; or pharmaceutically acceptable salts thereof, together or individually, constitute about 0.01 to 0.20 area percent of the HPLC chromatogram of a compound of formula I or a pharmaceutically acceptable salt thereof. In some embodiments, one or more compounds selected from I-1, I-2, I-3, I-4, I-5, I-6, or I-7; or pharmaceutically acceptable salts thereof, together or individually, are approximately 0.02-0.18, 0.03-0.16, 0.05-0.15, 0.075-0.13, 0.09-0.1, 0 .1-0.2 or 0.15-0.2 area percent of the HPLC chromatogram relative to the compound of formula I or a pharmaceutically acceptable salt thereof. In some embodiments, the above HPLC chromatogram area percentages are measured relative to the total area of the HPLC chromatogram and not relative to the peak area of a compound of formula I or a pharmaceutically acceptable salt thereof. In some embodiments, one or more compounds selected from I-1, I-2, I-3, I-4, I-5, I-6, or I-7; or pharmaceutically acceptable salts thereof, together or individually, constitute less than about 5.0 weight percent (% w/w), or about 0.01-5.0% w/w, relative to the compound of Formula I or a pharmaceutically acceptable compound thereof salt. In some embodiments, one or more compounds selected from I-1, I-2, I-3, I-4, I-5, I-6, or I-7; or pharmaceutically acceptable salts thereof

- 18 046788 individually or individually constitute less than about 3.0% w/w. compositions X4P-001 or are approximately 0.02-4.0, 0.03-3.5, 0.05-3.1, 0.05-2.9, 0.05-2.5, 0.05 -2.0, 0.05-1.8, 0.05-1.6, 0.05-1.5, 0.051.4, 0.05-1.2, 0.05-1.1, 0 .05-1.0, 0.05-0.9, 0.05-0.8, 0.05-0.7, 0.05-0.6, 0.05-0.5, 0.05 -0.4, 0.05-0.3, 0.05-0.2, 0.05-0.1 or about 0.1-0.5% w/w. compositions Х4Р-001.

In some embodiments, the total amount of organic impurities, including one or more compounds selected from I-1, I-2, I-3, I-4, I-5, I-6, or I-7; or pharmaceutically acceptable salts thereof, is less than about 0.05% w/w, about 0.1, 0.5, 1.0, 2.0, 3.0, or about 4.0% w/w . or less composition X4P-001. In some embodiments, the total amount of organic impurities is about 0.02-4.0, 0.03-3.5, 0.05-3.1, 0.05-2.9, 0.05-2.5, 0.05-2.0, 0.05-1.8, 0.05-1.6, 0.05-1.5, 0.05-1.4, 0.05-1.2, 0, 05-1.1, 0.05-1.0, 0.05-0.9, 0.05-0.8, 0.05-0.7, 0.05-0.6, 0.05- 0.5, 0.050.4, 0.05-0.3, 0.05-0.2, 0.05-0.1 or about 0.1-0.5% w/w. compositions Х4Р-001.

In some embodiments, the amount of I-1 or a pharmaceutically acceptable salt thereof is less than about 1.1%, 1%, 0.9%, 0.8%, 0.7%, 0.6%, 0.5%, 0.4%, 0.3%, 0.2%, 0.1%, 0.09%, 0.08%, 0.07%, 0.06%, 0.05%, 0.04%, 0.03%, 0.02% or 0.01% w/w compositions Х4Р-001. In some embodiments, I-1 or a pharmaceutically acceptable salt thereof is not detectable.

In some embodiments, the amount of I-2 or a pharmaceutically acceptable salt thereof is less than about 0.3%, 0.2%, 0.1%, 0.09%, 0.08%, 0.07%, 0.06 %, 0.05%, 0.04%, 0.03%, 0.02% or 0.01% w/w. compositions Х4Р-001.

In some embodiments, the amount of I-3 or a pharmaceutically acceptable salt thereof is less than about 0.4%, 0.3%, 0.2%, 0.1%, 0.09%, 0.08%, 0.07 %, 0.06%, 0.05%, 0.04%, 0.03%, 0.02% or 0.01% w/w. compositions Х4Р-001.

In some embodiments, the amount of I-4 or a pharmaceutically acceptable salt thereof is less than about 0.5%, 0.4%, 0.3%, 0.2%, 0.1%, 0.09%, 0.08 %, 0.07%, 0.06%, 0.05%, 0.04%, 0.03%, 0.02% or 0.01% w/w. compositions Х4Р-001. In some embodiments, I-4 or a pharmaceutically acceptable salt thereof is not detectable.

In some embodiments, the amount of I-5 or a pharmaceutically acceptable salt thereof is less than about 0.5%, 0.4%, 0.3%, 0.2%, 0.1%, 0.09%, 0.08 %, 0.07%, 0.06%, 0.05%, 0.04%, 0.03%, 0.02% or 0.01% w/w. compositions Х4Р-001.

In some embodiments, the amount of I-6 or a pharmaceutically acceptable salt thereof is less than about 0.5%, 0.4%, 0.3%, 0.2%, 0.1%, 0.09%, 0.08 %, 0.07%, 0.06%, 0.05%, 0.04%, 0.03%, 0.02% or 0.01% w/w. compositions Х4Р-001.

In some embodiments, the amount of I-7 or a pharmaceutically acceptable salt thereof is less than about 0.5%, 0.4%, 0.3%, 0.2%, 0.1%, 0.09%, 0.08 %, 0.07%, 0.06%, 0.05%, 0.04%, 0.03%, 0.02% or 0.01% w/w. compositions Х4Р-001.

In some embodiments, the amount of I-1 or a pharmaceutically acceptable salt thereof is from about 0.001 to about 1.1%, from about 0.01 to about 1.1%, from about 0.01 to about 0.8%, from 0 .01 to about 0.7%, from about 0.01 to about 0.6%, from 0.01 to about 0.5%, from 0.01 to about 0.4%, from 0.01 to about 0 .3%, from 0.01 to about 0.2%, from 0.01 to about 0.1%, from 0.01 to about 0.09%, from 0.01 to about 0.08%, from 0 .01 to about 0.07%, 0.01 to about 0.06%, 0.01 to about 0.05%, 0.01 to about 0.03%, or 0.01 to about 0. 02% w/w compositions Х4Р-001.

In some embodiments, the amount of I-2 or a pharmaceutically acceptable salt thereof is from about 0.001 to about 0.3%, from about 0.01 to about 0.3%, from 0.01 to about 0.2%, from 0. 01 to about 0.1%, 0.01 to about 0.09%, 0.01 to about 0.08%, 0.01 to about 0.07%, 0.01 to about 0.06 %, from 0.01 to about 0.05%, from 0.01 to about 0.03%, or from 0.01 to about 0.02% wt./wt. compositions Х4Р-001. In some embodiments, the amount of I3 or a pharmaceutically acceptable salt thereof is from about 0.001 to about 0.4%, from about 0.01 to about 0.4%, from about 0.01 to about 0.3%, from 0.01 to about 0.2%, from 0.01 to about 0.1%, from 0.01 to about 0.09%, from 0.01 to about 0.08%, from 0.01 to about 0.07% , from 0.01 to about 0.06%, from 0.01 to about 0.05%, from 0.01 to about 0.03%, or from 0.01 to about 0.02% w/w. compositions Х4Р-001.

In some embodiments, the amount of I-4 or a pharmaceutically acceptable salt thereof is from about 0.001 to about 0.5%, from about 0.01 to about 0.5%, from about 0.01 to about 0.3%, from 0 .01 to about 0.2%, from 0.01 to about 0.1%, from 0.01 to about 0.09%, from 0.01 to about 0.08%, from 0.01 to about 0, 07%, 0.01 to about 0.06%, 0.01 to about 0.05%, 0.01 to about 0.03%, or 0.01 to about 0.02% w/w . compositions Х4Р-001.

In some embodiments, the amount of I-5 or a pharmaceutically acceptable salt thereof is from about 0.001 to about 0.5%, from about 0.01 to about 0.5%, from about 0.01 to about 0.4%, from about 0.01 to about 0.3%, 0.01 to about 0.2%, 0.01 to about 0.1%, 0.01 to about 0.09%, 0.01 to about 0 .08%, 0.01 to about 0.07%, 0.01 to about 0.06%, 0.01 to about 0.05%, 0.01 to about 0.03%, or 0 .01 to about 0.02% w/w. compositions Х4Р-001. In some embodiments, the amount of I-6 or a pharmaceutical thereof

- 19 046788 commercially acceptable salt is from about 0.001 to about 0.5%, from about 0.01 to about 0.5%, from about 0.01 to about 0.4%, from about 0.01 to about 0, 3%, from 0.01 to about 0.2%, from 0.01 to about 0.1%, from 0.01 to about 0.09%, from 0.01 to about 0.08%, from 0. 01 to about 0.07%, 0.01 to about 0.06%, 0.01 to about 0.05%, 0.01 to about 0.03%, or 0.01 to about 0.02 % w/w compositions Х4Р-001. In some embodiments, the amount of I7 or a pharmaceutically acceptable salt thereof is from about 0.001 to about 0.5%, from about 0.01 to about 0.5%, from about 0.01 to about 0.4%, from about 0. 01 to about 0.3%, 0.01 to about 0.2%, 0.01 to about 0.1%, 0.01 to about 0.09%, 0.01 to about 0.08 %, 0.01 to about 0.07%, 0.01 to about 0.06%, 0.01 to about 0.05%, 0.01 to about 0.03%, or 0.01 to about 0.02% w/w. compositions Х4Р-001. In some embodiments, the amount of any additional or unknown impurities in the X4P-001 composition is from about 0.01 to about 0.2% w/w. the specified composition.

In some embodiments, the amount of n-hydroxybenzoic acid present in the X4P-001 composition is from about 0.01 to about 0.5% w/w. the specified composition. In some embodiments, the composition is substantially free of n-hydroxybenzoic acid. In some embodiments, p-hydroxybenzoic acid is not present in the composition in a detectable amount. In some embodiments, the X4P-001 composition contains a compound of Formula I or a pharmaceutically acceptable salt thereof and does not contain a compound of Formula I-4 or a pharmaceutically acceptable salt thereof in a detectable amount. In some embodiments, the chiral purity of the X4P-001 composition is at least about 97% enantiomeric excess (% ee). In some embodiments, the chiral purity of the compound of Formula I is at least 97% ee. In some embodiments, the chiral purity of the compound of Formula I is at least 98% ee. In some embodiments, the chiral purity of the compound of Formula I is at least 99% ee. In some embodiments, the chiral purity of the compound of formula I is at least 99.1, 99.2, 99.3, 99.4, 99.5, 99.6, 99.7, 99.8, or 99.9% ee .

In some embodiments, the present invention provides any compound described above and herein in isolated form. As used herein, the term isolated means that the connection is provided in a form that is separated from other components that may be present in the normal environment surrounding the connection. In some embodiments, the isolated compound is in solid form. In some embodiments, the isolated compound has at least about 50% purity as determined by a suitable HPLC method. In some embodiments, the isolated compound has a purity of at least about 60%, 70%, 80%, 90%, 95%, 98%, 99%, 99.5%, 99.6%, 99.7%, 99. 8%, 99.9%, 99.95%, 99.99% or 99.999%, determined by a suitable HPLC method. Expressed as a percentage, purity can be measured by weight percent of the desired compound (% w/w), by area % relative to the total area of the HPLC chromatogram, or by other methods known in the art.

Methods of preparation and analysis applicable to certain compounds of the present invention are described in US 7354934, WO 00/56729, USSN 60/232891 and USSN 60/234510, as well as in An, H.; Wang, T.; Mohan, V.; Griffey, R. H.; Cook, P. D. Tetrahedron 1998, 54, 3999-4012; the entire contents of each of which are incorporated herein by reference. The proposed compounds can be purified by any methods known in the art. Such methods include, for example, silica gel column chromatography; medium pressure liquid chromatography (MPLC); high performance liquid chromatography (HPLC); preparative HPLC (prepHPLC); flash chromatography (FC); liquid chromatography (LC); supercritical fluid chromatography (SFC); thin layer chromatography (TLC); preparative TLC (prep-TLC); liquid chromatography-mass spectrometry (LC-MS, LCMS or LC/MS); recrystallization; deposition; grinding into powder; distillation; derivatization; acid-base extraction and the like. The term purified, in purified form, or in isolated and purified form for a compound refers to the physical state of said compound after it has been isolated during synthesis (eg, from a reaction mixture), or from a natural source, or a combination thereof. Thus, the term purified, in purified form, or in isolated and purified form for a compound refers to the physical state of said compound (or a tautomer or stereoisomer thereof, or a pharmaceutically acceptable salt or solvate of said compound, said stereoisomer, or said tautomer) after its production in the process purification or by processes described in the present invention or well known to one skilled in the art (e.g., chromatography, recrystallization, and the like), in sufficient purity to be suitable for in vivo or medical use, and/or to be characterized using standard analytical methods described in the present invention or well known to one skilled in the art.

The term detectable amount as used herein refers to a component

- 20 046788 present in a sample, for example a sample of the proposed composition X4P-001, which is present at least in an amount that can be detected using analytical means known in the art. For example, in some embodiments, the detectable amount is at least an amount detectable by HPLC, LC-MS, mass spectrometry, NMR, or other analytical methods known to one skilled in the art or described in the present invention.

Mixtures of diastereomers can be separated into individual diastereomers based on their physicochemical differences by methods well known to those skilled in the art, such as, for example, chromatography and/or fractional crystallization. Enantiomers can be separated by converting a mixture of enantiomers into a mixture of diastereomers by reacting with a suitable optically active compound (e.g., a chiral auxiliary such as a chiral alcohol or Mosher acid chloride), separating the diastereomers, and converting (e.g., by hydrolysis) the individual diastereomers to the corresponding pure enantiomers. Enantiomers can also be separated using a chiral HPLC column.

4. Applications, preparation and administration

Pharmaceutically acceptable compositions

In one aspect of the present invention, there is provided a composition X4P-001 comprising a compound of the invention, a pharmaceutically acceptable salt or a pharmaceutically acceptable derivative thereof, or a composition of the invention X4P-001 and a pharmaceutically acceptable excipient, carrier, adjuvant or carrier medium. The amount of compound in the compositions of the present invention is such that it is effective to measurably inhibit CXCR4 or a mutant thereof in a biological sample or patient. In some embodiments, the amount of a compound in the compositions of the present invention is effective to measurably inhibit CXCR4 or a mutant thereof in a biological sample or patient. In some embodiments, a composition of the present invention is formulated for administration to a patient in need of such a composition. In some embodiments, the composition of the present invention is formulated for oral administration to a patient.

The term patient as used herein means an animal, preferably a mammal and most preferably a human. The term pharmaceutically acceptable carrier, adjuvant or transport medium refers to a non-toxic carrier, adjuvant or transport medium that does not neutralize the pharmacological activity of the compound with which it is formulated. Pharmaceutically acceptable carriers, adjuvants or transfer media that may be used in the compositions of the present invention include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins such as human serum albumin, buffering agents such as phosphates, glycine, sorbic acid, potassium sorbate, mixtures of partial glycerides of saturated vegetable fatty acids, water, salts or electrolytes such as protamine sulfate, sodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silicon dioxide, magnesium trisilicate, polyvinylpyrrolidone, substances based on cellulose, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, block copolymers of polyethylene and polyoxypropylene, polyethylene glycol and lanolin. Pharmaceutically acceptable derivative means any non-toxic salt, ester, ester salt or other derivative of a compound of the present invention capable of, when administered to a recipient, directly or indirectly, providing the compound of the present invention or an inhibitory active metabolite or residue thereof.

As used herein, the term inhibitory active metabolite or residue thereof means that the metabolite or residue of said compound is also an inhibitor of CXCR4 or a mutant thereof.

The compositions of the present invention can be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, vaginally, or through an implanted reservoir. The term parenteral as used herein includes subcutaneous, intravenous, intramuscular, intraarticular, intrasynovial, intrathoracic, intrathecal, intrahepatic, intralesional, and intracranial injection or infusion techniques. Preferably, said compositions are administered orally, intraperitoneally or intravenously. Sterile injectable forms of the compositions of the present invention may be an aqueous or oleaginous suspension. Said suspensions can be prepared according to methods known in the art, using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent, for example a solution in 1,3-butanediol. Acceptable transfer media and solvents that can be used include water, Ringer's solution, and isotonic sodium chloride solution. In addition, sterile fatty oils are traditionally used as a solvent or suspending medium. For this purpose, you can use any light fatty oil.

- 21 046788 lo, including synthetic mono- or diglycerides. Fatty acids such as oleic acid and its glyceride derivatives are suitable for use in the preparation of injectable formulations, as are natural pharmaceutically acceptable oils such as olive oil or castor oil, particularly in their polyoxyethylated variants. These oil solutions or suspensions may also contain a long chain alcohol diluent or dispersing agent, such as carboxymethylcellulose, or similar dispersing agents, which are widely used in the preparation of pharmaceutically acceptable dosage forms, including emulsions and suspensions. Other commonly used surfactants such as Tweens, Spans, and other emulsifying or bioavailability enhancing agents that are widely used in the manufacture of pharmaceutically acceptable solid, liquid, or other dosage forms may also be used for formulation purposes.

The pharmaceutically acceptable compositions of the present invention can be administered orally in any suitable oral dosage form, including, but not limited to, capsules, tablets, aqueous suspensions or solutions. For oral tablets, commonly used carriers include lactose and corn starch. Lubricants such as magnesium stearate are also commonly added. When administered orally in capsule form, suitable diluents include lactose and dried corn starch. When aqueous suspensions are required for oral administration, said active ingredient is combined with emulsifying and suspending agents. If necessary, some sweetening, flavoring or coloring agents may also be added.

Alternatively, the pharmaceutically acceptable compositions of the present invention can be administered in the form of suppositories for rectal administration. Said suppositories may be prepared by mixing said agent with a suitable non-irritating excipient which is solid at room temperature but liquid at rectal temperature and will thus melt in the rectum to release said drug. Such substances include cocoa butter, beeswax and polyethylene glycols.

Pharmaceutically acceptable compositions of the present invention can also be administered topically, particularly when the target of treatment includes areas or organs readily accessible for topical administration, including diseases of the eye, skin or lower intestinal tract. Suitable formulations for topical administration are readily available for each of the designated areas or organs.

Topical administration to the lower intestinal tract may be in a rectal suppository formulation (see above) or in a suitable enema formulation. Topical transdermal patches can also be used.

For topical applications, the pharmaceutically acceptable compositions of the invention may be provided in the form of a suitable ointment containing said active ingredient suspended or dissolved in one or more carriers. Carriers for topical administration of the compounds of the present invention include, but are not limited to, mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyoxyethylene, polyoxypropylene compound, emulsifying wax and water. Alternatively, the pharmaceutically acceptable compositions provided may be formulated in the form of a suitable lotion or cream containing said active ingredients suspended or dissolved in one or more pharmaceutically acceptable carriers. Suitable carriers include, but are not limited to, mineral oil, sorbitan monostearate, polysorbate 60, wax cetyl esters, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water.

For ophthalmic use, the pharmaceutically acceptable compositions of the invention may be formulated as micronized suspensions in sterile pH-adjusted isotonic saline or, preferably, as solutions in sterile isotonic pH-adjusted saline with or without a preservative such as benzalkonium chloride. Alternatively, for ophthalmic applications, these pharmaceutically acceptable compositions can be prepared in the form of an ointment such as petrolatum. Pharmaceutically acceptable compositions of the present invention can also be administered by nasal aerosol or inhalation. Such compositions are prepared according to methods well known in the art of preparing pharmaceutical formulations, and they can be prepared as saline solutions using benzyl alcohol or other suitable preservatives, absorption promoters to enhance bioavailability, fluorocarbons and/or other conventional solubilizing or dispersing agents .

Most preferably, the pharmaceutically acceptable compositions of the present invention are formulated for oral administration. Such formulations may be administered with or without food. In some embodiments, the pharmaceutically acceptable compositions of the present invention are administered without food. In other embodiments, the pharmaceutically acceptable compositions of the present invention are administered with food. The number of compounds of the present invention that can be combined with carrier substances to form a composition in

- 22 046788 single dose dosage form will vary depending on the subject being treated and the specific route of administration. Preferably, the proposed compositions should be formulated so that a dose of said inhibitor of 0.01-100 mg/kg body weight/day can be administered to a patient receiving said compositions. It should also be understood that the specific dosage and treatment regimen for any particular patient will depend on a number of factors, including the potency of the particular compound used, age, body weight, general health, gender, diet, time of administration, rate of elimination, drug combination , the assessment of the attending physician and the severity of the specific disease being treated. The amount of a compound of the present invention in a composition will also depend on the particular compound in said composition.

Compounds and compositions in accordance with the method of the present invention can be administered using any amount and any route of administration effective for treating or reducing the severity of cancer, autoimmune disorder, primary immunodeficiency, proliferative disorder, inflammatory disorder, neurodegenerative or neurological disorder, schizophrenia, disorder, bone-related, liver disease or heart disorder. The exact amount required will vary from subject to subject depending on the species, age and general condition of the subject, severity of the infection, the specific agent, route of administration, and the like. In some embodiments, the compounds of the present invention are formulated in unit dosage form for ease of administration and consistency of dosage.

The term patient or subject as used herein means an animal, preferably a mammal, and most preferably a human.

The pharmaceutically acceptable compositions of the present invention can be administered to humans and animals orally, rectally, parenterally, intracisternally, intravaginally, intraperitoneally, topically (in the form of powders, ointments or drops), buccally, in the form of an oral or nasal spray, or the like, depending on the severity infection being treated. In some embodiments, the compounds of the present invention may be administered orally or parenterally at a dosage of from about 0.01 to about 50 mg/kg, or, for example, from about 1 to about 25 mg/kg of subject body weight per day, one or more times per day. day to obtain the desired therapeutic effect.

Liquid dosage forms for oral administration include, but are not limited to, pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active compounds, these liquid dosage forms may contain inert diluents commonly used in the art, such as, for example, water or other solvents, solubilizing agents and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate , propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (particularly cottonseed, peanut, corn, germ, olive, castor and sesame oils), glycerin, tetrahydrofurfuryl alcohol, polyethylene glycols and sorbitan fatty acid esters, and mixtures of them. In addition to inert diluents, oral compositions may also contain adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring and odorant agents.

Injectable preparations, for example sterile aqueous or oleaginous injectable suspensions, can be prepared by methods known in the art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution, suspension or emulsion in a non-toxic parenterally acceptable diluent or solvent, for example a solution in 1,3-butanediol. Acceptable transfer media and solvents that may be used include water, USP Ringer's solution, and isotonic sodium chloride solution. In addition, sterile fatty oils are traditionally used as a solvent or suspending medium. For this purpose, any light fatty oil can be used, including synthetic mono or diglycerides. In addition, fatty acids such as oleic acid are used to prepare injection formulations.

Injectable formulations may be sterilized, for example, by filtration through a bacteria-retaining filter or by incorporating sterilizing agents in the form of sterile solid compositions that can be dissolved or dispersed in sterile water or other sterile injectable media prior to use.

To prolong the effect of a compound of the present invention, it is often desirable to slow down the absorption of the compound by subcutaneous or intramuscular injection. This can be achieved by using a liquid suspension of a crystalline or amorphous substance with poor water solubility. The rate of absorption of the specified compound in this case depends on the rate of its dissolution, which, in turn, may depend on the crystal size and crystalline form. As an alternative, delayed absorption of the parenterally administered form of the said compound is achieved by dissolving or suspending the said compound in oils

- 23 046788 new transfer medium. Depot injection forms are prepared by preparing microencapsulated matrices of the specified compound in biodegradable polymers such as polylactide-polyglycolide. Depending on the ratio of the compound to the polymer and the nature of the particular polymer used, the rate of release of the compound can be adjusted. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Injectable depot formulations are also prepared by incorporating the compound into liposomes or microemulsions that are compatible with body tissues.

Compositions for rectal or vaginal administration are preferably suppositories, which can be prepared by mixing the compounds of the present invention with suitable non-irritating excipients or carriers, such as cocoa butter, polyethylene glycol or suppository wax, which are solid at ambient temperature but liquid at body temperature and thus melt in the rectum or vaginal cavity and release the active compound.

Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In such solid dosage forms, the active compound is mixed with at least one inert pharmaceutically acceptable excipient or carrier, such as sodium citrate or calcium hydrogen phosphate, and/or a) fillers or extenders, such as starches, lactose, sucrose, glucose , mannitol and silicic acid, b) binders such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone, sucrose and gum arabic, c) humectants such as glycerin, d) leavening agents such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, some silicates and sodium carbonate, f) dissolution retarding agents such as paraffin, f) absorption accelerators such as quaternary ammonium compounds, g) wetting agents such as, for example, cetyl alcohol and monostearate glycerin, h) absorbents such as kaolin and bentonite clay, and i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof. In the case of capsules, tablets and pills, said dosage form may also contain buffering agents. Solid compositions of this type can also be used as fillers in soft and hard gelatin capsules using excipients such as lactose or milk sugar, as well as high molecular weight polyethylene glycols and the like. Solid dosage forms such as tablets, dragees, capsules, pills and granules can be prepared using coatings and coatings, such as enteric coatings and other coatings well known in the pharmaceutical formulation art. They may optionally contain opacifying agents and may also be formulated such that they release the active ingredient(s) only or predominantly in a specific part of the intestine, optionally in a delayed manner. Examples of encapsulating compositions that can be used include polymeric substances and waxes. Solid compositions of this type can also be used as fillers in soft and hard gelatin capsules using excipients such as lactose or milk sugar, as well as high molecular weight polyethylene glycols and the like.

Said active compounds may also be in microencapsulated form with one or more of the excipients mentioned above. Solid dosage forms such as tablets, dragees, capsules, pills and granules can be prepared using coatings and shells, such as enteric coatings, controlled release coatings and other coatings well known in the art of pharmaceutical formulations. In such solid dosage forms, the active compound may be mixed with at least one inert diluent such as sucrose, lactose or starch. Such dosage forms may also contain, as is common practice, additional substances other than inert diluents, such as tablet lubricants and other tableting excipients such as magnesium stearate and microcrystalline cellulose. In the case of capsules, tablets and pills, these dosage forms may also contain buffering agents. They may optionally contain opacifying agents and may also be formulated such that they release the active ingredient(s) only or predominantly in a specific part of the intestine, optionally in a delayed manner. Examples of encapsulating compositions that can be used include polymeric substances and waxes.

Dosage forms for topical or transdermal administration of a compound of the present invention include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants or patches. Said active ingredient is mixed under sterile conditions with a pharmaceutically acceptable carrier and any necessary preservatives or buffers that may be required. Ophthalmic composition, ear drops and eye drops are also included within the scope of the present invention. In addition, the present invention involves the use of transdermal patches, which have the additional advantage of providing controlled delivery of the compound to the body. Such dosage forms can be prepared using

- 24 046788 by dissolving or dispersing the specified compound in a suitable medium. Absorption enhancers can also be used to increase the flux of the compound through the skin. The rate can be controlled either by providing a rate-regulating membrane or by dispersing the compound in a polymer matrix or gel.

In some embodiments, the composition is formulated for oral administration in the form of a tablet or capsule. In some embodiments, the composition comprising X4P-001 is formulated for oral administration in capsule form.

In some embodiments, the proposed method includes administering to a patient one or more capsules containing from 10 to 1200 mg of the active ingredient X4P-001 and one or more pharmaceutically acceptable excipients. In some embodiments, said capsule consists of hard gelatin.

In some embodiments, the present invention provides a pharmaceutical composition comprising composition X4P-001, one or more diluents, a disintegrant, a lubricant, a flow agent, and a wetting agent. In some embodiments, the present invention provides a pharmaceutical composition containing from 10 to 1200 mg of composition X4P-001, microcrystalline cellulose, calcium hydrogen phosphate dihydrate, croscarmellose sodium, sodium stearyl fumarate, colloidal silicon dioxide and sodium lauryl sulfate. In some embodiments, the present invention provides a unit dosage form, wherein the unit dosage form contains a pharmaceutical composition containing 10,200 mg of composition X4P-001, microcrystalline cellulose, calcium hydrogen phosphate dihydrate, croscarmellose sodium, sodium stearyl fumarate, colloidal silicon dioxide and sodium lauryl sulfate. In some embodiments, the present invention provides a unit dosage form containing a pharmaceutical composition comprising composition X4P-001 present in an amount of about 10 mg, about 20 mg, about 25 mg, about 50 mg, about 75 mg, about 100 mg, about 150 mg, about 200 mg, about 250 mg, about 300 mg, about 400 mg, about 450 mg, about 500 mg, about 600 mg, about 700 mg, about 750 mg, about 800 mg, about 900 mg, about 1000 mg, about 1100 mg or about 1200 mg. In some embodiments, the proposed composition (or unit dosage form) is administered to a patient once daily, twice daily, three times daily, or four times daily. In some embodiments, the proposed composition (or unit dosage form) is administered to a patient once daily or twice daily.

In some embodiments, the present invention provides a pharmaceutical composition comprising:

(a) the proposed composition X4P-001, comprising approximately 30-40% by weight of the composition;

(b) microcrystalline cellulose in an amount of about 20-25% by weight of the composition;

(c) calcium hydrogen phosphate dihydrate in an amount of about 30-35% by weight of the composition;

(d) croscarmellose sodium in an amount of about 5-10% by weight of the composition;

(e) sodium stearyl fumarate in an amount of about 0.5-2% by weight of the composition;

(f) colloidal silica in an amount of about 0.1-1.0% by weight of the composition; and (g) sodium lauryl sulfate in an amount of about 0.1-1.0% by weight of the composition.

In some embodiments, the present invention provides a pharmaceutical composition comprising:

(a) the proposed composition X4P-001, comprising approximately 8-25% by weight of the composition;

(b) microcrystalline cellulose in an amount of about 65-85% by weight of the composition;

(c) croscarmellose sodium in an amount of about 2-10% by weight of the composition;

(d) sodium stearyl fumarate in an amount of about 0.1-3% by weight of the composition; and (e) colloidal silicon dioxide in an amount of about 0.05-0.7% by weight of the composition.

In some embodiments, the present invention provides a pharmaceutical composition comprising:

(a) the proposed composition X4P-001, comprising approximately 25-45% by weight of the composition;

(b) microcrystalline cellulose in an amount of about 10-35% by weight of the composition;

(c) calcium hydrogen phosphate dihydrate in an amount of about 15-45% by weight of the composition;

(d) croscarmellose sodium in an amount of about 2-10% by weight of the composition;

(e) sodium stearyl fumarate in an amount of about 0.3-2.5% by weight of the composition;

(f) colloidal silica in an amount of about 0.05-1.2% by weight of the composition; and (g) sodium lauryl sulfate in an amount of about 0.2-1.2% by weight of the composition.

In some embodiments, the present invention provides a pharmaceutical composition comprising:

(a) the proposed composition X4P-001, comprising approximately 35-75% by weight of the composition;

(b) microcrystalline cellulose in an amount of about 5-28% by weight of the composition;

(c) calcium hydrogen phosphate dihydrate in an amount of about 7-30% by weight of the composition;

(d) croscarmellose sodium in an amount of about 2-10% by weight of the composition;

- 25 046788 (e) sodium stearyl fumarate in an amount of approximately 0.3-2.5% by weight of the composition;

(f) colloidal silica in an amount of about 0.05-1.2% by weight of the composition; and (g) sodium lauryl sulfate in an amount of about 0.2-1.2% by weight of the composition.

In some embodiments, composition X4P-001 is present in an amount of about 10 mg, about 20 mg, about 25 mg, about 50 mg, about 75 mg, about 100 mg, about 150 mg, about 200 mg, about 250 mg, about 300 mg , about 400 mg, about 450 mg, about 500 mg, about 600 mg, about 700 mg, about 750 mg, about 800 mg, about 900 mg, about 1000 mg, about 1100 mg or about 1200 mg.

In some embodiments, the composition contains approximately 37 wt.% of the proposed composition X4P-001.

In some embodiments, the present invention provides a unit dosage form containing the proposed composition X4P-001 or a pharmaceutical composition.

In some embodiments, the present invention provides a dosage unit form comprising a pharmaceutical composition comprising:

(a) the proposed composition X4P-001 in an amount of approximately 10-30% by weight of the composition;

(b) microcrystalline cellulose in an amount of about 60-80% by weight of the composition;

(c) croscarmellose sodium in an amount of about 5-10% by weight of the composition;

(d) sodium stearyl fumarate in an amount of about 0.5-2% by weight of the composition; and (e) colloidal silicon dioxide in an amount of about 0.1-1.0% by weight of the composition.

In some embodiments, the present invention provides a dosage unit form containing a pharmaceutical composition comprising:

(a) the proposed composition X4P-001 in an amount of approximately 14.7% by weight of the composition;

(b) microcrystalline cellulose in an amount of about 78.1% by weight of the composition;

(c) croscarmellose sodium in an amount of about 6.0% by weight of the composition;

(d) sodium stearyl fumarate in an amount of about 1.0% by weight of the composition; and (e) colloidal silicon dioxide in an amount of about 0.2% by weight of the composition.

In some embodiments, the present invention provides a dosage unit form comprising a pharmaceutical composition comprising:

(a) the proposed composition X4P-001 in an amount of approximately 10-20% by weight of the composition;

(b) microcrystalline cellulose in an amount of about 25-40% by weight of the composition;

(c) calcium hydrogen phosphate dihydrate in an amount of about 35-55% by weight of the composition;

(d) croscarmellose sodium in an amount of about 4-15% by weight of the composition;

(e) sodium stearyl fumarate in an amount of about 0.3-2% by weight of the composition;

(f) colloidal silica in an amount of about 0.1-1.5% by weight of the composition; and (g) sodium lauryl sulfate in an amount of about 0.1-1.5% by weight of the composition.

In some embodiments, the present invention provides a dosage unit form comprising a pharmaceutical composition comprising:

(a) the proposed composition X4P-001 in an amount of approximately 12.85% by weight of the composition;

(b) microcrystalline cellulose in an amount of about 31.92% by weight of the composition;

(c) calcium hydrogen phosphate dihydrate in an amount of about 44.4% by weight of the composition;

(d) croscarmellose sodium in an amount of approximately 8.33% by weight of the composition;

(e) sodium stearyl fumarate in an amount of about 1.38% by weight of the composition;

(f) colloidal silica in an amount of about 0.42% by weight of the composition; and (g) sodium lauryl sulfate in an amount of about 0.7% by weight of the composition.

International Patent Application No. PCT/US 2016/066634 describes additional compositions and uses of X4P-001 and is incorporated herein by reference in its entirety.

Since it may be desirable to administer a combination of active compounds, for example, to provide treatment for a particular disease or condition, it is within the scope of the present invention that two or more pharmaceutical compositions, at least one of which contains a composition of the present invention, can be conveniently combined to in the form of a kit suitable for co-administration of said compositions. Thus, the kit according to the present invention contains two or more separate pharmaceutical compositions, at least one of which contains the composition X4P-001 according to the present invention, and means for holding said compositions separately from each other, such as a container, a divided bottle or divided into separate the foil bag. An example of such a set is the well-known blister pack used for packaging tablets, capsules and the like.

The kit of the present invention is particularly suitable for administering different dosage forms, for example oral and parenteral forms, for administering individual compositions at different dosage intervals, or for titrating individual compositions against each other. To facilitate treatment compliance, the kit usually contains instructions for administration and may be supplied with a memory aid.

- 26 046788

Uses of compounds and pharmaceutically acceptable compositions

It was found that during the synthesis of X4P-001, some impurities appear, such as the compounds listed above in table. 1 or their stereoisomers, or pharmaceutically acceptable salts. Isolating and characterizing each impurity is important for a number of reasons. Typically, pharmaceutical compositions require a high level of purity to meet regulatory standards for drug quality and purity. For example, the synthesis of X4P-001 typically produces impurities, including degradation products or production by-products, which may interfere with the therapeutic effects of X4P-001 and/or may be toxic if present in sufficient quantities. Accordingly, it is necessary to be able to determine the presence and quantities of such impurities and control the chemical purity, including stereochemical purity, of X4P-001. This requires the identification, isolation and chemical characterization of impurities that can be used in chromatographic procedures as standards to confirm the purity of X4P-001.

Accordingly, in one aspect of the present invention there is provided a method for preparing a compound of the invention or a pharmaceutically acceptable salt thereof, comprising contacting a suitable starting material or materials under the conditions set forth, for example, in the examples below, to produce the compound or a pharmaceutically acceptable salt thereof. In some embodiments, the compound or a pharmaceutically acceptable salt thereof is suitable for use as a reference standard and/or in methods for determining the presence of an impurity in a sample, such as a sample of a compound of Formula I or a pharmaceutically acceptable salt thereof.

The present invention also provides methods for determining an impurity comprising introducing a standard solution containing a compound of Formula I or a pharmaceutically acceptable salt thereof onto an HPLC column under a specified set of conditions to obtain a first HPLC chromatogram, wherein the specified amount and/or chemical entity of the compound present in the standard solution , are known; introducing a sample solution containing a compound of formula I or a pharmaceutically acceptable salt thereof onto an HPLC column under a specified set of conditions to obtain a second HPLC chromatogram; and determining the presence and/or amount of a compound in said sample solution. In some embodiments, said reference solution is administered multiple times. In some embodiments, the determination includes comparing the retention times of peaks in the first HPLC chromatogram and peaks in the second HPLC chromatogram to determine the presence of the compound in the sample solution. In other embodiments, the determination includes quantifying the peak areas of a sample solution and the peak areas of a reference solution in HPLC chromatograms and estimating from these results the amount of compound in the sample solution. In some embodiments, the HPLC column is a reverse phase column and the contents of the column are eluted using a mobile phase containing water, methanol, trifluoroacetic acid, or mixtures thereof.

The present invention also provides methods for determining the presence or amount of an impurity in a sample containing or essentially consisting of a compound of formula I or a pharmaceutically acceptable salt thereof, comprising introducing into an HPLC column, by one or more injections, a sample solution containing the material and bound ) with a reference compound having a known chemical structure, such as compound I-1, I-2, I-3, I-4, I-5, I-6 or I-7; obtaining an HPLC chromatogram and determining the presence and/or amount of a compound in the material. In some embodiments, the HPLC column is a reverse phase column and the contents of the column are eluted using a mobile phase containing water, methanol, trifluoroacetic acid, or mixtures thereof. The method may further include documenting in writing the chemical identity of the compound and the amount of the compound as an impurity. In some embodiments, the method further includes documenting in writing the chemical identity of the compound and the amount of the compound as an impurity in the material. In some cases, the amount of a compound in a material is determined by (i) identifying a peak in the chromatogram that corresponds to a peak in the control chromatogram of a compound known to have the structure I-1, I-2, I-3, I-4, I -5, I-6 or I-7; (ii) identifying the peak in the chromatogram that corresponds to the relative retention time of a compound known to have structure I-1, I-2, I-3, I-4, I-5, I-6 or I-7 ; and/or (iii) identifying a peak in the chromatogram that corresponds to a known amount of bound compound known to have the structure I-1, I-2, I-3, I-4, I-5, I-6, or I-7. In some embodiments, the HPLC column is a reverse phase column and the column is eluted using a mobile phase containing water, methanol, trifluoroacetic acid, or mixtures thereof. In some embodiments, the present invention provides a compound selected from I-1, I-2, I-3, I-4, I-5, I-6, or I7; or a pharmaceutically acceptable salt thereof, in a purity suitable for use as a reference or standard in various analytical methods (eg, HPLC, GC, SFC, LCMS), as more fully described below. In some embodiments, the compound or a pharmaceutically acceptable salt thereof may be isolated in at least 0.5% purity, at least 1% purity, pure

- 27 046788 that at least 5%, at least 10% pure, at least 15% pure, at least 25% pure, at least 50% pure, at least 75% pure, at least 95 pure % or a purity of at least 97%. In some embodiments, the compound or a pharmaceutically acceptable salt thereof is isolated and/or packaged as a solid.

In another aspect of the present invention, methods are provided for determining the presence and/or amount of I-1, I-2, I-3, I-4, I-5, I-6 or I-7; or pharmaceutically acceptable salts thereof. For example, the compound or a pharmaceutically acceptable salt thereof may be produced as an impurity during the synthesis of X4P-001. As used herein, the term impurity may refer to degradation products that are formed during storage of X4P-001 and/or by-products formed in the chemical reaction to produce X4P-001. In one embodiment, the method includes administering a reference solution containing I-1, I-2, I-3, I-4, I-5, I-6, or I-7; or pharmaceutically acceptable salts thereof, to an HPLC column under a set of conditions to obtain a first HPLC chromatogram, wherein said amount and/or chemical entity is I-1, I-2, I-3, I-4, I-5, I-6 or I-7; or pharmaceutically acceptable salts thereof present in the standard solution are known; injecting a sample solution containing X4P-001 into the HPLC column under the same set of conditions to obtain a second HPLC chromatogram and comparing the first HPLC chromatogram with the second HPLC chromatogram to determine the presence and/or amount of the impurity (I-1, I-2, I- 3, I-4, I-5, I-6 or I-7; or pharmaceutically acceptable salts thereof). The reference solution may be prepared by dissolving a sample (eg, a solid sample) of I-1, I-2, I-3, I-4, I-5, I-6, or I-7; or pharmaceutically acceptable salts thereof, in a first solvent, and said sample solution may be prepared by dissolving a sample of the solid in a second solvent. In some embodiments, the reference solution may contain additional compound(s), wherein the amount and/or nature of the additional compound(s) is also known. In one embodiment, the sample (eg, sample solution) may contain X4P-001. It should be understood that the present invention may include other samples that are expected to contain a compound selected from I-1, I-2, I-3, I-4, I-5, I-6 or I-7; or a pharmaceutically acceptable salt thereof.

In one embodiment, the presence of I-1, I-2, I-3, I-4, I-5, I-6, or I-7; or a pharmaceutically acceptable salt thereof in a sample solution can be determined by comparing the retention times of the peaks in the first HPLC chromatogram with the retention times of the peaks in the second HPLC chromatogram. For example, for a standard solution containing I-1, I-2, I-3, I-4, I-5, I-6, or I-7; or a pharmaceutically acceptable salt thereof, a chromatogram with a peak corresponding to I-1, I-2, I-3, I-4, I-5, I-6 or I-7 may be obtained; or a pharmaceutically acceptable salt thereof, and having a specific retention time. The sample solution can then be injected into the HPLC column under the same conditions as the standard solution, and the resulting chromatogram can be examined to determine whether it contains a peak at the same retention time as the peak corresponding to I-1, I-2, I-3, I-4, I-5, I-6 or I-7; or pharmaceutically acceptable salts thereof, in the HPLC chromatogram for the standard solution. The presence of such a peak may indicate that I-1, I-2, I-3, I-4, I-5, I-6 or I-7 are present in the sample; or a pharmaceutically acceptable salt thereof. In another embodiment, the amount is I-1, I-2, I-3, I-4, I-5, I-6, or I-7; or a pharmaceutically acceptable salt thereof in a sample solution can be determined by comparing the peak areas of the first HPLC chromatogram with the peak areas of the second HPLC chromatogram and calculating the content of I-1, I-2, I-3, I-4, I-5, I -6, or I-7; or a pharmaceutically acceptable salt thereof, in the sample solution.

Some embodiments of the present invention provide methods for determining an impurity in a material consisting essentially of X4P-001, wherein a sample solution containing the material and associated with a reference compound known to have the chemical structure I-1, I-2 , I-3, I-4, I-5, I-6, or I-7; or pharmaceutically acceptable salts thereof, as described in the present invention, are introduced into the HPLC column and an HPLC chromatogram is obtained to determine the presence and/or amount of the compound in the material. The methods of the present invention may further include documenting in writing the chemical nature of the compound and the amount of the compound as an impurity in the material. In other embodiments, the present invention provides methods for determining an impurity in a material consisting essentially of X4P-001, in which a solution of the material dissolved therein is injected onto an HPLC column and an HPLC chromatogram is obtained to determine the amount of the compound known to be present in the material. it has the structure I-1, I-2, I-3, I-4, I-5, I-6 or I-7; or pharmaceutically acceptable salts thereof, as described in the present invention. The chemical nature of the compound and the amount of the compound as an impurity in the material can then be documented. The amount of a compound in a material can be determined by (i) identifying a peak in the chromatogram that corresponds to a peak in the control chromatogram, (ii) identifying a peak in the chromatogram that corresponds to the relative retention time of a compound known to have structure I-1, I-2, I-3, I-4, I-5, I-6 or I-7; or pharmaceutically acceptable salts thereof, and/or (iii) identifying a peak in the chromatogram that corresponds to a known amount of bound compound known to have the structure I-1, I-2, I-3, I-4, I- 5, I-6, or I-7; or a pharmaceutically acceptable salt thereof.

Some embodiments of the present invention can be used to determine if

- 28 046788 quality and/or availability of I-1, I-2, I-3, I-4, I-5, I-6 or I-7; or pharmaceutically acceptable salts thereof, in a sample containing X4P-001. The sample may be freshly prepared material, or the sample may be a sample stored for a certain period of time. In one embodiment, a sample of X4P-001 may be stored and periodically analyzed using the methods described in the present invention to determine the presence and/or amount of I-1, I-2, I-3, I-4, I-5 , I-6, or I-7; or pharmaceutically acceptable salts thereof, in a sample that may be formed, for example, during the decomposition of X4P-001. In some cases, the sample may be placed under unfavorable conditions, that is, conditions specifically conducive to the degradation of X4P-001, such as elevated temperatures and/or increased humidity, and the sample is periodically analyzed using the methods described in the present invention to determine the presence of and/or amounts of I-1, I-2,I-3, I-4, I-5, I-6, or I-7; or pharmaceutically acceptable salts thereof. The compounds and compositions described in the present invention can be used to inhibit CXCR4 or a mutant thereof. Certain compounds and compositions described in the present invention have been found to be useful in the treatment, prevention and/or reduction of the risk of a disease, disorder or condition associated with CXCR4.

In one aspect of the present invention, there is provided a method of inhibiting the activity of CXCR4 or a mutant thereof in a patient, comprising the step of administering to said patient the proposed composition X4P-001. In other embodiments, the present invention provides a method of treating a disorder mediated by CXCR4 or a mutant thereof in a patient in need thereof, comprising the step of administering to said patient a composition of the present invention, X4P-001, or a pharmaceutically acceptable composition thereof. Such disorders are described in detail in the present invention.

Certain methods of treating a disease or disorder with X4P-001 are described in PCT Application No. PCT/US 2018/038776, filed June 21, 2018, the contents of which are incorporated herein by reference in their entirety. The compounds and compositions described herein can be used in such methods of treating a disease or disorder. In some embodiments, a composition comprising X4P-001 or a pharmaceutically acceptable salt thereof is administered orally in an amount of from about 200 to about 1200 mg per day. In some embodiments, the dosage composition may be administered twice daily in divided doses approximately 12 hours apart. In other embodiments, the dosage composition may be administered once daily. The terminal half-life of X4P-001 has been determined to generally be from about 12 to about 24 hours, or about 14.5 hours. The oral dose may be from about 100 to about 1200 mg once or twice daily. In some embodiments, the dose of X4P-0001 or a pharmaceutically acceptable salt thereof suitable for use in the present invention is from about 200 to about 800 mg per day. In other embodiments, the dose of X4P-001 or a pharmaceutically acceptable salt thereof suitable for use in the present invention may be from about 200 to about 600 mg, from about 400 to about 800 mg, from about 600 to about 1000 mg, or from about 800 up to approximately 1200 mg per day.

In one aspect, the present invention provides a method of treating cancer, such as described in the present invention, by administering to a patient in need thereof an effective amount of the disclosed composition X4P-001. In some embodiments, the method includes coadministering simultaneously or sequentially an effective amount of one or more additional therapeutic agents, such as those described herein. In some embodiments, the method includes co-administration with one additional therapeutic agent. In some embodiments, the method includes co-administration with two additional therapeutic agents. In some embodiments, the combination of a compound or composition of the invention and an additional therapeutic agent or agents acts synergistically to prevent or reduce cancer immune evasion and/or evasion of antiangiogenic therapy. In some embodiments, the patient has previously received another anticancer agent, such as adjuvant therapy or immunotherapy. In some embodiments, said cancer is refractory.

In some embodiments, the disease, disorder, or condition associated with CXCR4 is selected from cell proliferative disorders, Alzheimer's disease, HIV, rheumatoid arthritis, or pulmonary fibrosis. In some embodiments, the disease, disorder, or condition is a hyperproliferative disorder, such as cancer. In some embodiments, said cancer is breast, ovarian, kidney, lung, or melanoma. In some embodiments, the cancer is selected from renal cell carcinoma (RCC), refractory RCC, or clear cell RCC (ccRCC).

In some embodiments, the present invention provides a method of treating patients with cancer that is a solid tumor. In some embodiments, the patient has kidney cancer, a renal tumor, renal carcinoma (including clear cell and papillary renal carcinoma), ovarian cancer, or melanoma.

- 29 046788

The compounds of the invention are CXCR4 inhibitors and are thus suitable for use in the treatment of one or more disorders associated with CXCR4 activity. Thus, in some embodiments, the present invention provides a method of treating a CXCR4-mediated disorder, comprising the step of administering to a patient in need thereof the disclosed composition X4P-001 or a pharmaceutically acceptable composition thereof.

In one aspect of the present invention, there is provided a method of treating cancer in a patient in need thereof, wherein said method comprises administering to said patient the disclosed composition X4P-001 in combination with one or more additional therapeutic agents, such as one or more immunostimulating therapeutic compounds.

In some embodiments, one or more immunostimulatory therapeutic compounds is selected from elotuzumab, mifamurtide, a toll-like receptor agonist or activator, or a RORYt activator.

In some embodiments, the method further includes administering to said patient a third therapeutic agent, such as an immune checkpoint inhibitor. In some embodiments, the method includes administering to a patient in need thereof three therapeutic agents selected from the disclosed composition X4P-001, an immunostimulatory therapeutic compound, and an immune checkpoint inhibitor.

In some embodiments, the immune checkpoint inhibitor is selected from nivolumab, pembrolizumab, ipilimumab, avelumab, durvalumab, atezolizumab, or pidilizumab.

In another aspect of the present invention, there is provided a method of treating cancer in a patient in need thereof, wherein said method comprises administering to said patient the disclosed composition X4P-001 in combination with one or more additional therapeutic agents selected from an indoleamine (2,3) dioxygenase (IDO) inhibitor ), poly(ADP-ribose) polymerase (PARP) inhibitor, histone deacetylase (HDAC) inhibitor, CDK4/CDK6 inhibitor, or phosphatidylinositol 3-kinase (PI3K) inhibitor.

In some embodiments, the IDO inhibitor is selected from epacadostat, indoximod, capmanitib, GDC-0919, PF-06840003, BMC:F001287, Phy906/KD108, or an enzyme that degrades kynurenine.

In some embodiments, the PARP inhibitor is selected from olaparib, rucaparib, or niraparib.

In some embodiments, the HDAC inhibitor is selected from vorinostat, romidepsin, panobinostat, belinostat, entinostat, or chidamide.

In some embodiments, the CDK 4/6 inhibitor is selected from palbociclib, ribociclib, abemaciclib, or trilaciclib.

In some embodiments, the method further includes administering to said patient a third therapeutic agent, such as an immune checkpoint inhibitor. In some embodiments, the method includes administering to a patient in need thereof three therapeutic agents selected from the proposed composition X4P-001, a second therapeutic agent selected from an indoleamine (2,3) dioxygenase (IDO) inhibitor, a poly(ADP-ribose) inhibitor, polymerase (PARP), a histone deacetylase (HDAC) inhibitor, a CDK4/CDK6 inhibitor or a phosphatidylinositol 3-kinase (PI3K) inhibitor; and a third therapeutic agent selected from an immune checkpoint inhibitor. In some embodiments, the immune checkpoint inhibitor is selected from nivolumab, pembrolizumab, ipilimumab, avelumab, durvalumab, atezolizumab, or pidilizumab.

In some embodiments, the PI3K inhibitor is selected from idelalisib, alpelisib, taselisib, pictilisib, copanlisib, duvelisib, PQR309, or TGR1202.

In another aspect of the present invention, there is provided a method of treating cancer in a patient in need thereof, wherein said method comprises administering to said patient the disclosed composition X4P-001 in combination with one or more additional therapeutic agents selected from platinum-based therapeutic agents, a taxane, a nucleoside inhibitor, or a therapeutic agent that interferes with normal DNA synthesis, protein synthesis, cell replication, or will otherwise inhibit rapidly proliferating cells.

In some embodiments, the platinum-based therapeutic agent is selected from cisplatin, carboplatin, oxaliplatin, nedaplatin, picoplatin, or satraplatin.

In some embodiments, the taxane is selected from paclitaxel, docetaxel, albumin-bound paclitaxel, cabazitaxel, or SID530. In some embodiments, the therapeutic agent that interferes with normal DNA synthesis, protein synthesis, cell replication, or would otherwise interfere with the replication of rapidly proliferating cells is selected from trabectedine, mechlorethamine, vincristine, temozolomide, cytarabine, lomustine, azacitidine, omacetaxine mepesuccinate, asparaginase Erwinia chrysanthemi, eribulin mesylate, capacetrin, bendamustine, ixabepilone, nelarabine, clorafabine, trifluridine or tipiracil.

In some embodiments, the method further includes administering to said patient

- 30 046788 third therapeutic agent, such as an immune checkpoint inhibitor. In some embodiments, the method includes administering to a patient in need thereof three therapeutic agents selected from the proposed composition X4P-001, a second therapeutic agent selected from a platinum-based therapeutic agent, a taxane, a nucleoside inhibitor, or a therapeutic agent that interferes with normal DNA synthesis, protein synthesis , cell replication or otherwise inhibiting rapidly proliferating cells, and a third therapeutic agent selected from an immune checkpoint inhibitor. In some embodiments, the immune checkpoint inhibitor is selected from nivolumab, pembrolizumab, ipilimumab, avelumab, durvalumab, atezolizumab, or pidilizumab.

In some embodiments, any one of the above methods further includes the step of obtaining a biological sample from the patient and determining the amount of a disease-associated biomarker. In some embodiments, the biological sample is a blood sample.

In some embodiments, the disease-associated biomarker is selected from circulating CD8+ T cells or a CD8+ T cell:Treg cell ratio.

In some embodiments, the cancer is selected from hepatocellular carcinoma, ovarian cancer, ovarian epithelial cancer, fallopian tube cancer; papillary serous cystadenocarcinoma or uterine papillary serous carcinoma (UPSC); prostate cancer; testicular cancer; gallbladder cancer; hepatocholangiocarcinoma; synovial sarcoma of soft tissues and bone; rhabdomyosarcomas; osteosarcomas; chondrosarcomas; Ewing's sarcoma; anaplastic thyroid cancer; adrenocortical adenoma; pancreatic cancer; pancreatic ductal carcinoma or pancreatic adenocarcinoma; gastrointestinal/stomach cancer (GIST); lymphomas; squamous cell carcinoma of the head and neck (SCCHN); salivary gland cancer; glioma or brain cancer; neurofibromatosis-1 associated malignant peripheral nerve sheath tumors (MPNST); Waldenström's macroglobulinemia or medulloblastoma.

In some embodiments, the cancer is selected from hepatocellular carcinoma (HCC), hepatoblastoma, colon cancer, rectal cancer, ovarian cancer, ovarian epithelial cancer, fallopian tube cancer, papillary serous cystadenocarcinoma, uterine papillary serous carcinoma (UPSC), hepatocholangiocarcinoma, synovial sarcoma soft tissue and bone, rhabdomyosarcoma, osteosarcoma, anaplastic thyroid cancer, adrenocortical adenoma, pancreatic cancer, pancreatic ductal carcinoma, pancreatic adenocarcinoma, glioma, neurofibromatosis-1 associated malignant peripheral nerve sheath tumors (MPNST), Waldenström's macroglobulinemia or medulloblastoma .

In some embodiments, the present invention provides a method of treating a cancer that is a solid tumor, such as a sarcoma, carcinoma, or lymphoma, comprising the step of administering the disclosed composition X4P-001 to a patient in need thereof. Solid tumors typically contain an abnormal mass of tissue that usually does not include cysts or areas of fluid. In some embodiments, the cancer is selected from renal cell carcinoma or kidney cancer; hepatocellular carcinoma (HCC) or hepatoblastoma or liver cancer; melanoma; breast cancer; colorectal carcinoma or colorectal cancer; colon cancer; rectal cancer; anal cancer; lung cancer such as non-small cell lung cancer (NSCLC); small cell lung cancer (SCLC); ovarian epithelial cancer; ovarian carcinoma; fallopian tube cancer; papillary serous cystadenocarcinoma or uterine papillary serous carcinoma (UPSC); prostate cancer; testicular cancer; bladder cancer; hepatocholangiocarcinoma; synovial sarcoma of soft tissues and bone; rhabdomyosarcomas; osteosarcomas; chondrosarcomas; Ewing's sarcoma; anaplastic thyroid cancer; adrenocortical carcinoma; pancreatic cancer; pancreatic ductal carcinoma; pancreatic adenocarcinoma; gastrointestinal/stomach cancer (GIST); lymphomas; squamous cell carcinoma of the head and neck (SCCHN); salivary gland cancer; glioma or brain cancer; neurofibromatosis-1 associated malignant peripheral nerve sheath tumors (MPNST); Waldenström's macroglobulinemia or medulloblastoma.

In some embodiments, the cancer is selected from renal cell carcinoma, hepatocellular carcinoma (HCC), hepatoblastoma, colorectal carcinoma, colorectal cancer, colon cancer, colon cancer, anal cancer, ovarian cancer, ovarian epithelial cancer, ovarian carcinoma, fallopian cancer tubes, papillary serous cystadenocarcinoma, uterine papillary serous carcinoma (UPSC), hepatocholangiocarcinoma, synovial sarcoma of soft tissue and bone, rhabdomyosarcoma, osteosarcoma, chondrosarcoma, anaplastic thyroid cancer, adrenocortical carcinoma, pancreatic cancer, pancreatic ductal carcinoma, omas of the pancreas, glioma, brain cancer, neurofibromatosis-1 associated malignant peripheral nerve sheath tumors (MPNST), Waldenström's macroglobulinemia or medulloblastoma. In some embodiments, the cancer is selected from hepatocellular carcinoma (HCC), hepatoblastoma, colon cancer, rectal cancer, ovarian cancer,

- 31 046788 ovarian epithelial cancer, ovarian carcinoma, fallopian tube cancer, papillary serous cystadenocarcinoma, uterine papillary serous carcinoma (UPSC), hepatocholangiocarcinoma, synovial sarcoma of soft tissue and bone, rhabdomyosarcoma, osteosarcoma, anaplastic thyroid cancer, adrenocortical al carcinoma, pancreatic cancer , pancreatic ductal carcinoma, pancreatic adenocarcinoma, glioma, neurofibromatosis-1 associated malignant peripheral nerve sheath tumors (MPNST), Waldenström's macroglobulinemia, or medulloblastoma.

In some embodiments, the cancer is hepatocellular carcinoma (HCC). In some embodiments, the cancer is a hepatoblastoma. In some embodiments, the cancer is colon cancer. In some embodiments, the cancer is colorectal cancer. In some embodiments, the cancer is ovarian cancer or ovarian carcinoma. In some embodiments, the cancer is an ovarian epithelial cancer. In some embodiments, the cancer is fallopian tube cancer. In some embodiments, the cancer is papillary serous cystadenocarcinoma. In some embodiments, the cancer is uterine papillary serous carcinoma (UPSC). In some embodiments, the cancer is hepatocholangiocarcinoma. In some embodiments, the cancer is a synovial sarcoma of soft tissue and bone. In some embodiments, the cancer is a rhabdomyosarcoma. In some embodiments, the cancer is osteosarcoma. In some embodiments, the cancer is anaplastic thyroid cancer. In some embodiments, the cancer is adrenocortical carcinoma. In some embodiments, the cancer is pancreatic cancer or pancreatic ductal carcinoma. In some embodiments, the cancer is pancreatic adenocarcinoma. In some embodiments, the cancer is a glioma. In some embodiments, the cancer is a malignant peripheral nerve sheath tumor (MPNST). In some embodiments, the cancer is neurofibromatosis-1 associated MPNST. In some embodiments, the cancer is Waldenström's macroglobulinemia. In some embodiments, the cancer is a medulloblastoma. In some embodiments, the present invention provides a method of treating a cancer selected from leukemia or blood cancer, comprising administering to a patient in need thereof an effective amount of the disclosed composition X4P-001, optionally in combination with an additional therapeutic agent, such as the agents described herein. In some embodiments, the cancer is selected from acute myeloid leukemia (AML), chronic myeloid leukemia (CML), acute lymphocytic leukemia (ALL), chronic lymphocytic leukemia (CLL), or virus-induced leukemia.

In some embodiments, the patient has an operable solid tumor. This means that the patient's tumor can be removed surgically. In other embodiments, the patient has an inoperable solid tumor, in which the patient's tumor is considered not amenable to surgical removal, in whole or in part.

In some embodiments, the cancer is an advanced cancer, such as advanced kidney cancer or advanced renal cell carcinoma.

In some embodiments, the present invention provides a method of treating refractory cancer in a patient in need thereof, comprising administering to the patient in need thereof an effective amount of the disclosed composition X4P-001 or a pharmaceutical composition thereof, optionally in combination with an additional therapeutic agent such as described herein.

In some embodiments, the patient has previously been administered a protein kinase inhibitor. In some embodiments, the patient has previously been administered a VEGF-R antagonist. In some embodiments, the patient has previously been administered an immune checkpoint inhibitor. In some embodiments, the patient has previously been administered an immune checkpoint inhibitor selected from nivolubam (Opdivo®, Bristol-Myers Squibb), pembrolizumab (Keytruda®, Merck), or ipilimumab (Yervoy®, BristolMyers Squibb).

In some embodiments, the proposed composition X4P-001 is administered to a patient on an empty stomach.

Cell proliferative disorders

The present invention provides methods and compositions for diagnosing and prognosticating cell proliferative disorders (eg, cancer) and treating such disorders by targeting CXCR4.

Cellular proliferative disorders described in the present invention include, for example, cancer, obesity and proliferation-related diseases.

Such disorders can be diagnosed using methods known in the art.

Cancer

Cancer includes, in one embodiment, but is not limited to, leukemias (e.g., acute leukemia

- 32 046788 goats, acute lymphocytic leukemia, acute myelocytic leukemia, acute myelocytic leukemia, acute promyelocytic leukemia, acute myelomonocytic leukemia, acute monocytic leukemia, acute erythroleukemia, chronic leukemia, chronic myelocytic leukemia, chronic lymphocytic leukemia goats), polycythemia vera, lymphoma (eg , Hodgkin's disease or non-Hodgkin's lymphoma), Waldenström's macroglobulinemia, multiple myeloma, heavy chain disease, and solid tumors such as sarcomas and carcinomas (eg, fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteogenic sarcoma, chordoma, angiosarcoma, endothelial sarcoma, lymphangiosarcoma, oendothelial sarcoma, synovioma, mesothelioma, Ewing's tumor, leiomyosarcoma, rhabdomyosarcoma, colon carcinoma, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous carcinoma, papillary carcinoma , papillary adenocarcinoma, cystadenocarcinoma, medullary carcinoma, bronchogenic carcinoma, renal cell carcinoma, hepatoma, bile duct carcinoma, choriocarcinoma, seminoma, embryonal carcinoma, Wilms tumor, cervical cancer, uterine cancer, testicular cancer, lung carcinoma, small cell lung carcinoma, bladder carcinoma, epithelial carcinoma, glioma, astrocytoma, glioblastoma multiforme (GBM, also known as glioblastoma), medulloblastoma, craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, schwannoma, neurofibrosarcoma, meningioma, chalk anoma, neuroblastoma and retinoblastoma).

In some embodiments, the cancer is a glioma, astrocytoma, glioblastoma multiforme (GBM, also known as glioblastoma), medulloblastoma, craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, schwannoma, neurofibrosarcoma, meningioma, melanoma, neuroblastoma, or . In some embodiments, the cancer is an acoustic neuroma, an astrocytoma (e.g., grade I pilocytic astrocytoma, grade II low-grade astrocytoma, grade III anaplastic astrocytoma, or grade IV glioblastoma (GBM)), chordoma, CNS lymphoma, craniopharyngioma, glioma brainstem, ependymoma, mixed glioma, optic nerve glioma, subependymoma, medulloblastoma, meningioma, metastatic brain tumor, oligodendroglioma, pituitary tumors, primitive neuroectodermal (PNET) tumor, or schwannoma. In some embodiments, the cancer is a type of cancer more common in children than in adults, such as a brainstem glioma, craniopharyngioma, ependymoma, juvenile pilocytic astrocytoma (JPA), medulloblastoma, optic nerve glioma, pineal tumor, primitive neuroectodermal tumors ( PNET) or rhabdoid tumor. In some embodiments, the patient is an adult. In some embodiments, the patient is a child or pediatric patient.

In another embodiment, cancer includes, but is not limited to, mesothelioma, hepatobiliary cancer (liver and bile duct cancer), bone cancer, pancreatic cancer, skin cancer, head or neck cancer, cutaneous or intraocular melanoma, ovarian cancer, colon cancer, cancer rectal cancer, anal cancer, stomach cancer, gastrointestinal cancer (stomach cancer, colorectal cancer, duodenal cancer), uterine cancer, fallopian tube carcinoma, endometrial carcinoma, cervical carcinoma, vaginal carcinoma, vulvar carcinoma, Hodgkin's disease, esophageal cancer, small intestine cancer, endocrine system cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, prostate cancer, testicular cancer, chronic or acute leukemia, chronic myeloid leukemia, lymphocytic lymphomas, bladder cancer, kidney or ureteral cancer, renal cell carcinoma, renal pelvic carcinoma, non-Hodgkin's lymphoma, spinal axis tumors, brainstem glioma, pituitary adenoma, adrenocortical cancer, bladder cancer, multiple myeloma, cholangiocarcinoma , fibrosarcoma, neuroblastoma, retinoblastoma, or a combination of one or more of the above cancers.

The present invention further provides methods and compositions for the diagnosis, prognosis and treatment of virus-associated cancers, including immunodeficiency virus (HIV)-associated solid tumors, human papillomavirus (HPV)16 positive incurable solid tumors and adult T-cell leukemia, which is caused by human T-cell leukemia virus type I (HTLV-I) and is a highly aggressive form of CD4+ T-cell leukemia characterized by clonal integration of HTLV-I in leukemia cells (see https://clinicaltrials.gov/ct2/show/study/NCT02631746 ); and virus-associated tumors in gastric cancer, nasopharyngeal carcinoma, cervical cancer, vaginal cancer, vulvar cancer, head and neck squamous cell carcinoma, and Merkel cell carcinoma (see https://clinicaltrials.gov/ct2/show/study/ NCT02488759; see also https://clinicaltrials.gov/ct2/show/study/NCT0240886; https://clinicaltrials.gov/ct2/show/NCT02426892).

In some embodiments, the present invention provides a method of treating a tumor in a patient in need thereof, comprising administering to the patient the disclosed composition X4P-001. In some embodiments, the tumor includes any of the cancers described in the present invention

- 33 046788 ni. In some embodiments, the tumor includes a melanoma-type cancer. In some embodiments, the cancer includes breast cancer. In some embodiments, the tumor includes lung cancer. In some embodiments, the tumor includes small cell lung cancer (SCLC). In some embodiments, the tumor includes non-small cell lung cancer (NSCLC).

In some embodiments, the tumor is treated by stopping further growth of the tumor. In some embodiments, a tumor is treated by reducing the size (e.g., volume or weight) of the tumor by at least 5%, 10%, 25%, 50%, 75%, 90%, or 99% relative to the size of the tumor before treatment. In some embodiments, tumors are treated by reducing the number of tumors in the patient by at least 5%, 10%, 25%, 50%, 75%, 90%, or 99% relative to the number of tumors before treatment.

Primary immunodeficiency

In some embodiments, the present invention provides a method for treating one or more disorders, diseases and/or conditions, wherein said disorder, disease or condition includes, but is not limited to, a primary immunodeficiency disease or disorder, comprising administering to a patient in need thereof an effective amount of a compositions Х4Р-001. Primary immunodeficiencies treatable by the methods of the present invention include: warts, hypogammaglobulinemia, infections, myelocathexys syndrome (WHIM); severe congenital neutropenia (SCN), especially when caused by G6PC3 deficiency (McDermott et al. (2010) Blood 116:2793-2802); GATA2 deficiency (MonoMAC syndrome) (Maciejweski-Duval et al. (2015) J. Leukoc. Biol. 5MA0815-288R (electronic publication ahead of print); idiopathic CD4' T-lymphocytopenia (ICL) and Wiskott-Aldrich syndrome.

In other embodiments, the present invention provides a method of inhibiting CXCR4 activity in a biological sample, comprising the step of contacting said biological sample with a composition of the invention, X4P-001.

In accordance with another embodiment, the present invention relates to a method of inhibiting the activity of CXCR4 or a mutant thereof in a biological sample, comprising the step of contacting said biological sample with the proposed composition X4P-001. In some embodiments, the present invention provides a method of inhibiting the activity of CXCR4 or a mutant thereof in a biological sample, comprising the step of contacting said biological sample with a composition of the invention, X4P-001.

The term biological sample as used herein includes, without limitation, cell cultures or extracts thereof; biopsy material obtained from a mammal, or extracts thereof; and blood, saliva, urine, feces, semen, tears or other body fluids or extracts thereof.

Co-Administration with Additional Therapeutic Agents In one aspect of the present invention, there is provided a method of treating cancer, such as the cancers described herein, by administering the disclosed composition X4P-001 to a patient in need thereof. In some embodiments, the method comprises co-administering, simultaneously or sequentially, an effective amount of one or more additional therapeutic agents, such as the additional therapeutic agents described herein. In some embodiments, the method includes coadministration of one additional therapeutic agent. In some embodiments, the method includes coadministration of two additional therapeutic agents. In some embodiments, the combination of the proposed X4P-001 composition and an additional therapeutic agent or agents acts synergistically to prevent or reduce cancer immune evasion and/or evasion of antiangiogenic therapy. In some embodiments, the patient has previously received another anticancer agent, such as adjuvant therapy or immunotherapy. In some embodiments, said cancer is refractory.

Certain methods of treating a disease or disorder by coadministering X4P-001 with one or more additional agents are described in PCT Application No. PCT/US 2018/038776, filed June 21, 2018, the contents of which are incorporated herein by reference in their entirety.

Depending on the particular condition or disease being treated, additional therapeutic agents that are typically used to treat that condition may also be present in the compositions of the present invention. As used herein, additional therapeutic agents that are typically used to treat a particular disease or condition are referred to as appropriate for the disease or condition being treated.

In some embodiments, the additional therapeutic agent is a kinase inhibitor or a VEGF-R antagonist. Approved VEGF inhibitors and kinase inhibitors suitable for use in the present invention include: bevacizumab (Avastin®, Genentech/Roche), an anti-VEGF monoclonal antibody; ramucirumab (Cyramza®, Eli Lilly), an anti-VEGFR-2 antibody, and ziv-aflibercept, also known as VEGF Trap (Zaltrap®; Regeneron/Sanofi). VEGFR inhibitors such as regorafenib (Stivarga®, Bayer); vandetanib (Caprelsa®, AstraZeneca); axitinib (Inlyta®, Pfizer) and lenvatinib (Lenvima®, Eisai); Raf inhibitors such as sorafenib (Nexavar®, Bayer AG and Onyx);

- 34 046788 dabrafenib (Tafinlar®, Novartis) and vemurafenib (Zelboraf®, Genentech/Roche); MEK inhibitors such as cobimetinib (Cotellic®, Exelixis/Genentech/Roche); trametinib (Mekinist®, Novartis); Bcr-Abl tyrosine kinase inhibitors such as imatinib (Gleevec®, Novartis); nilotinib (Tasigna®, Novartis); dasatinib (Sprycel®, BristolMyersSquibb); bosutinib (Bosulif®, Pfizer) and ponatinib (Iclusig®, Ariad Pharmaceuticals); Her2 and EGFR inhibitors such as gefitinib (Iressa®, AstraZeneca); erlotinib (Tarceva®, Genentech/Roche/Astellas); lapatinib (Tykerb®, Novartis); afatinib (Gilotrif®, Boehringer Ingelheim); osimertinib (targeting EGFR with activating mutations, Tagrisso®, AstraZeneca) and brigatinib (Alunbrig®, Ariad Pharmaceuticals); c-Met and VEGFR2 inhibitors such as cabozantinib (Cometriq®, Exelixis); and multikinase inhibitors such as sunitinib (Sutent®, Pfizer); pazopanib (Votrient®, Novartis); ALK inhibitors such as crizotinib (Xalkori®, Pfizer); ceritinib (Zykadia®, Novartis) and alectinib (Alecenza®, Genentech/Roche); Bruton's tyrosine kinase inhibitors such as ibrutinib (Imbruvica®, Pharmacyclics/Janssen); and Flt3 receptor inhibitors such as midostaurin (Rydapt®, Novartis).

Other kinase inhibitors and VEGF-R antagonists that are under development and may be used in the present invention include tivozanib (Aveo Pharmaecuticals); vatalanib (Bayer/Novartis); lucitanib (Clovis Oncology); dovitinib (TKI258, Novartis); Chiauanib (Chipscreen Biosciences); CEP-11981 (Cephalon); linifanib (Abbott Laboratories); neratinib (HKI-272, Puma Biotechnology); radotinib (Supect®, IY5511, II-Yang Pharmaceuticals, South Korea); ruxolitinib (Jakafi®, Incyte Corporation); PTC299 (PTC Therapeutics); CP-547,632 (Pfizer); foretinib (Exelixis, GlaxoSmithKline); quizartinib (Daiichi Sankyo) and motesanib (Amgen/Takeda).

In some embodiments, the additional therapeutic agent is an mTOR inhibitor that inhibits cell proliferation, angiogenesis, and glucose uptake. Approved mTOR inhibitors suitable for use in the present invention include everolimus (Afinitor®, Novartis); temsirolimus (Torisel®, Pfizer) and sirolimus (Rapamune®, Pfizer).

In some embodiments, the additional therapeutic agent is a poly(ADP-ribose) polymerase (PARP) inhibitor. Approved PARP inhibitors suitable for use in the present invention include olaparib (Lynparza®, AstraZeneca); rucaparib (Rubraca®, Clovis Oncology) and niraparib (Zejula®, Tesaro). Other investigational PARP inhibitors that may be used in the present invention include talazoparib (MDV3800/BMN 673/LT00673, Medivation/Pfizer/Biomarin); veliparib (ABT-888, AbbVie) and BGB-290 (BeiGene, Inc.).

In some embodiments, the additional therapeutic agent is a phosphatidylinositol 3-kinase (PI3K) inhibitor. Approved PI3K inhibitors suitable for use in the present invention include idelalisib (Zydelig®, Gilead). Other investigational PI3K inhibitors that may be used in the present invention include alpelisib (BYL719, Novartis); tazelisib (GDC-0032, Genentech/Roche); pictilisib (GDC-0941, Genentech/Roche); copanlisib (BAY806946, Bayer); duvelisib (formerly IPI-145, Infinity Pharmaceuticals); PQR309 (Piqur Therapeutics, Switzerland) and TGR1202 (formerly RP5230, TG Therapeutics). In some embodiments, the additional therapeutic agent is a proteasome inhibitor. Approved proteasome inhibitors suitable for use in the present invention include bortezomib (Velcade®, Takeda); carfilzomib (Kyprolis®, Amgen) and ixazomib (Ninlaro®, Takeda).

In some embodiments, the additional therapeutic agent is a histone deacetylase (HDAC) inhibitor. Approved HDAC inhibitors suitable for use in the present invention include vorinostat (Zolinza®, Merck); romidepsin (Istodax®, Celgene); panobinostat (Farydak®, Novartis) and belinostat (Beleodaq®, Spectrum Pharmaceuticals). Other investigational HDAC inhibitors that can be used in the present invention include entinostat (SNDX-275, Syndax Pharmaceuticals) (NCT00866333) and chidamide (Epidaza®, HBI-8000, Chipscreen Biosciences, China).

In some embodiments, the additional therapeutic agent is a CDK inhibitor, such as a CDK 4/6 inhibitor. Approved CDK 4/6 inhibitors suitable for use in the present invention include palbociclib (Ibrance®, Pfizer) and ribociclib (Kisqali®, Novartis). Other investigational CDK 4/6 inhibitors that may be used in the present invention include abemaciclib (Ly2835219, Eli Lilly) and trilaciclib (G1T28, G1 Therapeutics).

In some embodiments, the additional therapeutic agent is an indoleamine (2,3)-dioxygenase (IDO) inhibitor. Investigational IDO inhibitors that can be used in the present invention include epacadostat (INCB024360, Incyte); indoximod (NLG-8189, NewLink Genetics Corporation); capmatinib (INC280, Novartis); GDC-0919 (Genentech/Roche); PF-06840003 (Pfizer); BMS:F001287 (Bristol-Myers Squibb); Phy906/KD108 (Phytoceutica) and kynurenine degrading enzyme (Kynase, Kyn Therapeutics).

In some embodiments, the additional therapeutic agent is a growth factor antagonist, such as a platelet-derived growth factor (PDGF) antagonist, or an epidermal

- 35 046788 growth factor (EGF) or its receptor (EGFR). Approved PDGF antagonists that can be used in the present invention include olaratumab (Lartruvo®; Eli Lilly). Approved EGFR antagonists that can be used in the present invention include cetuximab (Erbitux®, Eli Lilly); necitumumab (Portrazza®, Eli Lilly), panitumumab (Vectibix®, Amgen), and osimertinib (targeting EGFR with activating mutations, Tagrisso®, AstraZeneca). In some embodiments, the additional therapeutic agent is an aromatase inhibitor. Approved aromatase inhibitors that can be used in the present invention include exemestane (Aromasin®, Pfizer); anastrozole (Arimidex®, AstraZeneca) and letrozole (Femara®, Novartis).

In some embodiments, the additional therapeutic agent is a Hedgehog pathway antagonist. Approved Hedgehog signaling pathway inhibitors that can be used in the present invention include sonidegib (Odomzo®, Sun Pharmaceuticals) and vismodegib (Erivedge®, Genentech), which are indicated for the treatment of basal cell carcinoma.

In some embodiments, the additional therapeutic agent is a folic acid inhibitor. Approved folic acid inhibitors suitable for use in the present invention include pemetrexed (Alimta®, Eli Lilly).

In some embodiments, the additional therapeutic agent is a CC chemokine receptor 4 (CCR4) inhibitor.

Investigational CCR4 inhibitors that may be suitable for use in the present invention include mogamulizumab (Poteligeo®, Kyowa Hakko Kirin, Japan).

In some embodiments, the additional therapeutic agent is an isocitrate dehydrogenase (IDH) inhibitor. Investigational IDH inhibitors that can be used in the present invention include AG120 (Celgene; NCT 02677922); AG221 (Celgene, NCT 02677922; NCT 02577406); BAY1436032 (Bayer, NCT 02746081); IDH305 (Novartis, NCT 02987010). In some embodiments, the additional therapeutic agent is an arginase inhibitor. Investigational arginase inhibitors that may be used in the present invention include AEB1102 (PEGylated recombinant arginase, Aeglea Biotherapeutics), which is in Phase 1 clinical studies for the treatment of acute myeloid leukemia and myelodysplastic syndrome (NCT 02732184) and solid tumors (NCT 02561234); and SV-1158 (Calithera Biosciences). In some embodiments, the additional therapeutic agent is a glutaminase inhibitor. Investigational glutaminase inhibitors that may be used in the present invention include CB-839 (Calithera Biosciences).

In some embodiments, the additional therapeutic agent is an antibody that binds to tumor antigens, which are proteins expressed on the surface of tumor cells. Approved antibodies that bind to tumor antigens and can be used in the present invention include rituximab (Rituxan®, Genentech/Biogenldec); ofatumumab (anti-CD20 antibody, Arzerra®, GlaxoSmithKline); obinutuzumab (anti-CD20, Gazyva®, Genentech), ibritumomab (anti-CD20 with yttrium-90, Zevalin®, Spectrum Pharmaceuticals); daratumumab (anti-CD38 antibody, Darzalex®, Janssen Biotech), dinutuximab (anti-glycolipid GD2 antibody, Unituxin®, United Therapeutics); trastuzumab (anti-HER2 antibody, Herceptin®, Genentech); adotrastuzumab emtansine (anti-HER2 antibody condensed with emtansine, Kadcyla®, Genentech) and pertuzumab (anti-HER2 antibody, Perjeta®, Genentech); and brentuximab vedotin (anti-CD30 antibody-drug conjugate, Adcetris®, Seattle Genetics).

In some embodiments, the additional therapeutic agent is a topoisomerase inhibitor. Approved topoisomerase inhibitors that can be used in the present invention include irinotecan (Onivyde®, Merrimack Pharmaceuticals); topotecan (Hycamtin®, GlaxoSmithKline). Investigational topoisomerase inhibitors that can be used in the present invention include pixantrone (Pixuvri®, CTI Biopharma).

In some embodiments, the additional therapeutic agent is a nucleoside inhibitor or other therapeutic agent that interferes with normal DNA synthesis, protein synthesis, cell replication, or will otherwise inhibit rapidly proliferating cells. Such nucleoside inhibitors or other therapeutic agents include trabectedin (guanidine alkylating agent, Yondelis®, Janssen Oncology), mechlorethamine (alkylating agent, Valchlor®, Aktelion Pharmaceuticals); vincristine (Oncovin®, Eli Lilly; Vincasar®, Teva Pharmaceuticals; Marqibo®, Talon Therapeutics); temozolomide (a prodrug for the alkylating agent 5-(3methyltriazen-1-yl)-imidazol-4-carboxamide (MTIC) Temodar®, Merck); injectable form of cytarabine (ara-C, an anti-metabolic analogue of cytidine, Pfizer); lomustine (alkylating agent, CeeNU®, Bristol-Myers Squibb; Gleostine®, NextSource Biotechnology); azacitidine (pyrimidine nucleoside analogue of cytidine, Vidaza®, Celgene); omacetaxine mepesuccinate (cephalotaxin ester) (ingi

- 36 046788 protein synthesis inhibitor, Synribo®; Teva Pharmaceuticals); Erwinia chrysanthemi asparaginase (asparagine reducing enzyme, Elspar®, Lundbeck; Erwinaze®, EUSA Pharma); eribulin mesylate (microtubule inhibitor, tubulin-based antimitotic, Halaven®, Eisai); cabazitaxel (microtubule inhibitor, tubulin-based antimitotic, Jevtana®, Sanofi-Aventis); capacetrin (thymidylate synthase inhibitor, Xeloda®, Genentech); bendamustine (a bifunctional derivative of mechlorethamine that is predicted to form interstrand DNA cross-links, Treanda®, Cephalon/Teva); ixabepilone (semi-synthetic analogue of epothilone B, microtubule inhibitor, tubulin-based antimitotic, Ixempra®, Bristol-Myers Squibb); nelarabine (deoxyguanosine analogue prodrug, nucleoside metabolic inhibitor, Arranon®, Novartis); clofarabine (ribonucleotide reductase inhibitor prodrug, deoxycytidine competitive inhibitor, Clolar®, Sanofi-Aventis); and trifluridine and tipiracil (thymidine-based nucleoside analogue and thymidine phosphorylase inhibitor, Lonsurf®, Taiho Oncology). In some embodiments, the additional therapeutic agent is a platinum-based therapeutic agent, also referred to as platinum. Platinums cause DNA cross-linking and thereby inhibit DNA repair and/or DNA synthesis mainly in rapidly reproducing cells such as cancer cells. Approved platinum-based therapeutic agents that can be used in the present invention include cisplatin (Platinol®, Bristol-Myers Squibb); carboplatin (Paraplatin®, Bristol-Myers Squibb; and Teva; Pfizer); oxaliplatin (Eloxitin® Sanofi-Aventis) and nedaplatin (Aqupla®, Shionogi). Other platinum-based therapeutic agents that have undergone clinical trials and may be used in the present invention include picoplatin (Poniard Pharmaceuticals) and satraplatin (JM-216, Agennix).

In some embodiments, the additional therapeutic agent is a taxane compound that causes disruption of microtubules that are essential for cell division. Approved taxane compounds that can be used in the present invention include paclitaxel (Taxol®, Bristol-Myers Squibb), docetaxel (Taxotere®, Sanofi-Aventis; Docefrez®, Sun Pharmaceutical), albumin-bound paclitaxel (Abraxane®; Abraxis/Celgene ) and cabazitaxel (Jevtana®, Sanofi-Aventis). Other taxane compounds that have been clinically tested and may be used in the present invention include SID530 (SK Chemicals, Co.) (NCT 00931008). In some embodiments, the additional therapeutic agent is an inhibitor of anti-apoptotic proteins, such as BCL-2. Approved anti-apoptotic protein inhibitory agents that can be used in the present invention include venetoclax (Venclexta®, AbbVie/Genentech) and blinatumomab (Blincyto®, Amgen). Other therapeutic agents targeting anti-apoptotic proteins that have undergone clinical trials and may be used in the present invention include navitoclax (ABT-263, Abbott), a BCL-2 inhibitor (NCT 02079740).

The term checkpoint inhibitor as used herein refers to agents useful for preventing cancer cells from escaping the immune system of a patient. One of the main mechanisms of destabilization of the immune system against tumor is known as T-cell exhaustion, which occurs due to chronic exposure to antigens, resulting in up-regulation of inhibitory receptors. These inhibitory receptors serve as immune checkpoints to prevent uncontrolled immune reactions.

PD-1 and coinhibitory receptors such as cytotoxic T lymphocyte-associated antigen 4 (CTLA-4, B and T lymphocyte attenuator (BTLA; CD272), T-cell immunoglobulin and mucin domain-containing protein 3 (Tim-3) , lymphocyte activation gene 3 (Lag-3; CD223) and others are often called checkpoint regulators. They act as molecular gatekeepers that allow extracellular information to dictate whether cell cycle progression and other intracellular signal transduction processes should proceed.

In one aspect, the checkpoint inhibitor is a biological therapeutic agent or small molecule. In another aspect, the checkpoint inhibitor is a monoclonal antibody, a humanized antibody, a fully human antibody, a fusion protein, or a combination thereof. In an additional aspect, the checkpoint inhibitor inhibits a checkpoint protein selected from CTLA-4, PDL1, PDL2, PD1, B7-H3, B7-H4, BTLA, HVEM, TIM3, GAL9, LAG3, VISTA, KIR, 2B4, CD160, CGEN -15049, CHK1, CHK2, A2aR, B-7 family ligands, or combinations thereof. In an additional aspect, the checkpoint inhibitor interacts with a checkpoint protein ligand selected from CTLA-4, PDL1, PDL2, PD1, B7-H3, B7-H4, BTLA, HVEM, TIM3, GAL9, LAG3, VISTA, KIR, 2B4, CD160 , CGEN-15049, CHK1, CHK2, A2aR, B-7 family ligands, or combinations thereof. In one aspect, the checkpoint inhibitor is an immune stimulating agent, T cell growth factor, interleukin, antibody, vaccine, or a combination thereof. In a further aspect, the interleukin is IL-7 or IL-15. In a particular aspect, interleukin is glycosylated IL-7. In an additional aspect, the vaccine is a dendritic cell (DC) vaccine.

Checkpoint inhibitors include any agent that blocks or inhibits inhibitory pathways of the immune system in a statistically significant manner. Such inhibitors may include

- 37 046788 themselves small molecule inhibitors or may include antibodies, or antigen binding fragments thereof, that bind to and block or inhibit immune checkpoint receptors, or antibodies that bind to and block or inhibit ligands of immune checkpoint receptors. Exemplary checkpoint molecules that may be specific to block or inhibit include, but are not limited to, CTLA-4, PDL1, PDL2, PD1, B7-H3, B7-H4, BTLA, HVEM, GAL9, LAG3, TIM3, VISTA, KIR, 2B4 (belongs to the CD2 family of molecules and is expressed on all NK, γδ and memory CD8+ (αβ) T cells, CD160 (also called BY55), CGEN-15049, CHK1 and CHK2 kinases, A2aR and various B-7 family ligands. The B7 family of ligands includes, but is not limited to, B7-1, B7-2, B7-DC, B7-H1, B7-H2, B7-H3, B7-H4, B7-H5, B7-H6, and B7-H7. Checkpoint inhibitors include antibodies or antigen-binding fragments thereof, other binding proteins, biological therapeutics, or small molecules that bind to and block or inhibit the activity of one or more of CTLA-4, PDL1, PDL2, PD1, BTLA, HVEM, TIM3, GAL9 , LAG3, VISTA, KIR, 2B4, CD 160 and CGEN-15049 Exemplary immune checkpoint inhibitors include tremelimumab (CTLA-4 blocking antibody), anti-OX40, monoclonal antibody PD-L1 (anti-B7-H1; MEDI4736), MK-3475 (PD-1 blocker), nivolumab (Opdivo®, BMS-936558; anti-PD1 antibody), ST-011 (anti-PD1 antibody), BY55 monoclonal antibody, AMP224 (anti-PDL1 antibody), BMS-936559 (anti-PDL1 antibody), MPLDL3280A (anti-PDL1 antibody), MSB0010718C (anti-PDL1 antibody), and ipilimumab (anti-CTLA-4 checkpoint inhibitor). Checkpoint protein ligands include, but are not limited to, PD-L1, PD-L2, B7H3, B7-H4, CD28, CD86 and TIM-3.

In some embodiments, the immune checkpoint inhibitor is selected from a PD-1 antagonist, a PD-L1 antagonist, or a CTLA-4 antagonist. In some embodiments, a CXCR4 antagonist, such as X4P-001 or a pharmaceutically acceptable salt thereof, is administered in combination with nivolumab (anti-PD-1 antibody, Opdivo®, Bristol-Myers Squibb); pembrolizumab (anti-PD-1 antibody, Keytruda®, Merck); ipilimumab (anti-CTLA-4, Yervoy®, Bristol-Myers Squibb); durvalumab (anti-PD-L1, Imfinzi®, AstraZeneca) or atezolizumab (anti-PD-L1, Tecentriq®, Genentech).

Other immune checkpoint inhibitors suitable for use in the present invention include REGN2810 (Regeneron), an anti-PD-1 antibody tested in patients with basal cell carcinoma (NCT 03132636); NSCLC (NCT 03088540); squamous cell carcinoma of the skin (NCT02760498); lymphoma (NCT02651662) and melanoma (NCT 03002376); pidilizumab (CureTech), also known as CT-011, an antibody that binds to PD-1, being studied in clinical trials for diffuse large B-cell lymphoma and multiple myeloma; avelumab (Bavencio®, Pfizer/Merck KGaA, also known as MSB0010718C), a fully human IgG1 antibody to PD-L1, being studied in clinical trials in non-small cell lung cancer, Merkel cell carcinoma, mesothelioma, solid tumors, kidney cancer, ovarian cancer, bladder cancer, head and neck cancer and stomach cancer; and PDR001 (Novartis), an inhibitory antibody that binds to PD-1, being studied in clinical trials in non-small cell lung cancer, melanoma, triple-negative breast cancer and advanced or metastatic solid tumors. Tremelimumab (CP-675.206; Astrazeneca) is a fully human monoclonal antibody to CTLA-4 that has been studied in clinical trials in a number of indications, including: mesothelioma, colorectal cancer, kidney cancer, breast cancer, lung cancer and non-small cell lung cancer. ductal adenocarcinoma of the pancreas, pancreatic cancer, germ cell tumor, squamous cell carcinoma of the head and neck, hepatocellular carcinoma, prostate cancer, endometrial cancer, metastatic liver cancer, liver cancer, large B-cell lymphoma, ovarian cancer, cervical cancer, metastatic anaplastic cancer thyroid, urothelial cancer, fallopian tube cancer, multiple myeloma, bladder cancer, soft tissue sarcoma and melanoma. AGEN-1884 (Agenus) is an anti-CTLA4 antibody being studied in a phase I clinical trial in advanced solid tumors (NCT 02694822). Nivolumab (Opdivo®, BMS-93568/MDX1106; Bristol-Myers Squibb) is a fully human IgG4 monoclonal antibody that acts as an immunomodulator by binding to the programmed cell death 1 (PD-1) receptor and selectively blocking its interaction with PD-ligands. L1 and PD-L2. The structure and other properties of nivolumab are presented at http://www.drugbank.ca/drugs/DB09035, from which data were retrieved on March 14, 2016, and are incorporated herein. Nivolumab is approved for the treatment of patients with advanced renal cell carcinoma who have received prior antiangiogenic therapy; as a single agent for certain types of unresectable or metastatic melanoma; for the treatment of unresectable or metastatic melanoma or in combination with ipilimumab for the treatment of unresectable or metastatic melanoma; and for the treatment of metastatic non-small cell lung cancer that has progressed during or after platinum-based chemotherapy. In addition, nivolumab has been tested or has been mentioned as a potential

- 38 046788 means of treatment for other oncological indications, including solid tumors; skin melanoma; glioblastoma; glioma; gliosarcoma; astrocytoma; brain cancer; leukemia; acute myeloid leukemia; chronic myeloid leukemia; chronic lymphocytic leukemia; advanced liver cancer or hepatocellular carcinoma; uveal melanoma; prostate cancer; pancreatic neoplasm and pancreatic cancer; bladder cancer; colorectal cancer; myelodysplastic syndrome; Hodgkin's lymphoma; non-Hodgkin's lymphoma; multiple myeloma; cervical cancer; endometrial cancer; uterine cancer; ovarian cancer and ovarian carcinoma; peritoneal carcinoma; squamous cell carcinoma of the head and neck; stomach cancer; esophageal carcinoma; Kaposi's sarcoma; breast neoplasm, breast adenocarcinoma and breast cancer; bone sarcoma; soft tissue sarcoma; meningiomas and mesothelioma. In a phase 3 study of more than 800 patients with advanced clear cell renal cell carcinoma and who had been previously treated with one or two antiangiogenic regimens, patients were randomized to receive nivolumab 3 mg/kg intravenously. every two weeks or 10 mg everolimus orally in tablet form daily. Patients treated with nivolumab had a longer median overall survival, reduced hazard ratio for death, and higher objective response rate than patients treated with nivolumab (25%) compared with patients treated with everolimus (5%) (P < 0.001), with lower incidence of treatment-related grade 3 or 4 adverse events (Motzer et al. (2015), New England Journal of Medicine, 373:1803-1813). Accordingly, some embodiments of the present invention provide a method of treating advanced clear cell renal cell carcinoma, comprising administering to a patient in need thereof an effective amount of a CXCR4 antagonist, such as X4P-001 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof in combination with nivolumab or everolimus, at this patient has not necessarily received prior treatment with an antiangiogenic therapy regimen. In general, the amount of nivolumab or other immune checkpoint inhibitor useful in the present invention will depend on the size, weight, age and condition of the patient being treated, the severity of the disorder or condition, and the judgment of the treating physician. For example, current prescribing guidelines for the treatment of unresectable or metastatic renal cell carcinoma recommend nivolumab 3 mg/kg intravenous infusion over 60 minutes every two weeks until disease progression or unacceptable toxicity is detected. At the discretion of the treating physician, depending on individual tolerability, the prescribed dose of nivolumab may be increased, for example, by increasing the administered dose and/or frequency of administration. At the discretion of the attending physician, along with the warnings provided in the instructions for use, nivolumab may be discontinued or its dose may be reduced if significant adverse effects occur. In some embodiments, nivolumab is administered using the methods of the present invention in accordance with the above recommendations for use. In some embodiments, the present invention provides a method of treating a patient by administering a CXCR4 antagonist, such as X4P-001 or a pharmaceutically acceptable salt thereof, in combination with immunostimulating therapeutic agents. Approved immunostimulatory therapeutic agents that can be used in the present invention include elotuzumab (anti-SLAMF7 antibody, Empliciti®, Bristol-Myers Squibb). Investigational immunostimulatory compounds that may be used in the present invention include mifamurtide (Mepact®, Takeda Oncology). Another immunostimulatory therapeutic agent that can be used in the present invention is recombinant human interleukin 15 (rhIL-15). rhIL-15 has been tested in clinical practice as a treatment for melanoma and renal cell carcinoma (NCT 01021059 and NCT 01369888), as well as leukemia (NCT 02689453). Another immunostimulatory therapeutic agent that can be used in the present invention is recombinant human interleukin 12 (rhIL-12). Another suitable IL-15 immunotherapy is heterodimeric IL-15 (hetIL-15, Novartis/Admune), a hybrid complex consisting of a synthetic form of endogenous IL-15 complexed with a soluble IL-15 binding protein, IL receptor alpha chain -15 (IL15:sIL-15RA), which has been tested in phase I clinical trials in melanoma, renal cell carcinoma, non-small cell lung cancer, and head and neck squamous cell carcinoma (NCT 02452268). Recombinant human interleukin 12 (rhIL-12) has been tested in clinical practice in many oncology indications, for example, as a therapy for lymphoma (NM-IL-12, Neumedicines, Inc.), (NCT 02544724 and NCT 02542124). Another approach to stimulate the immune response is the use of oncolytic viruses. In some embodiments, the present invention provides a method of treating a patient by administering the disclosed composition X4P-001 in combination with an immunostimulating therapeutic agent, such as oncolytic viruses. Approved immunostimulating oncolytic viruses that can be used in the present

- 39 046788 invention include talimogene laherparepvec (live attenuated herpes simplex virus, Imlygic®, Amgen).

The term aromatase inhibitor as used herein refers to a compound that inhibits the production of estrogens, eg the conversion of the substrates androstenedione and testosterone to estrone and estradiol, respectively. The term includes, but is not limited to, steroids, in particular atamestane, exemestane and formestane, and in particular non-steroidal compounds, in particular aminoglutethimide, rogletimide, pyridoglutetimide, trilostane, testolactone, ketoconazole, vorozole, fadrozole, anastrozole and letrozole. Exemestane is marketed under the trade name Aromasin™. Formestane is marketed under the trade name Lentaron™. Fadrozole is marketed under the trade name Afema™. Anastrozole is marketed under the trade name Arimidex™. Letrozole is marketed under the trade names Femara™ or Femar™. Aminoglutethimide is marketed under the trade name Orimeten™. The combination according to the present invention containing a chemotherapeutic agent that is an aromatase inhibitor is particularly suitable for the treatment of hormone receptor-positive tumors such as breast tumors.

The term antiestrogen as used herein refers to a compound that antagonizes the effect of estrogens at the estrogen receptor level. The term includes, but is not limited to, tamoxifen, fulvestrant, raloxifene and raloxifene hydrochloride. Tamoxifen is marketed under the trade name Nolvadex™. Raloxifene hydrochloride is marketed under the trade name Evista™. Fulvestrant is available for administration under the trade name Faslodex™. The combination of the present invention containing a chemotherapeutic agent that is an antiestrogen is particularly suitable for use in the treatment of estrogen receptor positive tumors such as breast tumors.

The term antiandrogen as used herein refers to any substance that is capable of inhibiting the biological effects of androgenic hormones and includes, but is not limited to, bicalutamide (Casodex™). As used herein, the term gonadorelin agonist includes, but is not limited to, abarelix, goserelin and goserelin acetate. Goserelin is available for administration under the trade name Zoladex™.

The term topoisomerase I inhibitor as used herein includes, but is not limited to, topotecan, hymatecan, irinotecan, camptothecin and analogs thereof, 9-nitrocamptothecin and the macromolecular camptothecin conjugate PNU-166148. Irinotecan is available for administration, for example, in the form in which it is marketed, for example, under the brand name Camptosar™. Topotecan is marketed under the trade name Hycamptin™.

The term topoisomerase II inhibitor as used herein includes, but is not limited to, anthracyclines such as doxorubicin (including a liposomal formulation such as Caelyx™), daunorubicin, epirubicin, idarubicin and nerubicin, anthraquinones, mitoxantrone and losoxantrone, and the podophyllotoxins etoposide and teniposide. Etoposide is marketed under the trade name Etopophos™. Teniposide is marketed under the trade name VM 26-Bristol. Doxorubicin is marketed under the trade name Acriblastin™ or Adriamycin™. Epirubicin is marketed under the trade name Farmorubicin™. Idarubicin is marketed under the trade name Zavedos™. Mitoxantrone is marketed under the trade name Novantron.

The term microtubule-active agent refers to microtubule-stabilizing, microtubule-destabilizing, and microtubule polymerization inhibitor compounds, including, but not limited to, taxanes such as paclitaxel and docetaxel; vinca alkaloids such as vinblastine or vinblastine sulfate, vincristine or vincristine sulfate and vinorelbine; discodermolides; colchicine and epothilones and their derivatives. Paclitaxel is marketed under the trade name Taxol™. Docetaxel is marketed under the trade name Taxotere™. Vinblastine sulfate is marketed under the trade name Vinblastin R.P™. Vincristine sulfate is marketed under the trade name Farmistin™.

The term alkylating agent as used herein includes, but is not limited to, cyclophosphamide, ifosfamide, melphalan or nitrosourea (BCNU or Gliadel). Cyclophosphamide is marketed under the trade name Cyclostin™. Ifosfamide is marketed under the trade name Holoxan™.

The term histone deacetylase inhibitors or HDAC inhibitors in the present invention refers to compounds that inhibit histone deacetylase and which have antiproliferative activity. These include, but are not limited to, suberoylanilide hydroxamic acid (SAHA).

The term antitumor antimetabolite as used herein includes, but is not limited to, 5-fluorouracil or 5-FU, capecitabine, gemcitabine, DNA demethylating compounds such as 5-azacytidine and decitabine, methotrexate and edatrexate, and folic acid antagonists such as

- 40 046788 pemetrexed. Capecitabine is marketed under the trade name Xeloda™. Gemcitabine is marketed under the trade name Gemzar™. The term platinum compound in the present invention includes, but is not limited to, carboplatin, cisplatin, cisplatin and oxaliplatin. Carboplatin can be administered, for example, in the form in which it is marketed, for example, under the trade name Carboplat™. Oxaliplatin is available for administration, for example, in the form in which it is marketed, for example, under the trade name Eloxatin™.

In the present invention, the term compounds that target/reduce the activity of protein kinase or lipid kinase; or protein phosphatase or lipid phosphatase activity; or other anti-angiogenic compounds, includes, but is not limited to, protein tyrosine kinase inhibitors, and/or serine and/or threonine kinase inhibitors or lipid kinase inhibitors, such as a) compounds that target, reduce or inhibit the activity of platelet-derived growth factor receptors (PDGFR) , such as compounds that target, reduce or inhibit the activity of PDGFR, in particular compounds that inhibit the TRF receptor, such as an H-phenyl-2-pyrimidinamine derivative such as imatinib, SU101, SU6668 and GFB-111; b) compounds that target, reduce or inhibit the activity of fibroblast growth factor receptors (FGFR); c) compounds that target, reduce or inhibit insulin-like growth factor receptor I (IGF-IR) activity, such as compounds that target, reduce or inhibit IGF-IR activity, in particular compounds that inhibit IGF receptor I kinase activity , or antibodies that target the extracellular domain of IGF receptor I or its growth factors; d) compounds that target, reduce or inhibit the activity of members of the Trk receptor tyrosine kinase family, or ephrin B4 inhibitors; f) compounds that target, reduce or inhibit the activity of members of the Axl family of receptor tyrosine kinases; f) compounds that target, reduce or inhibit the activity of Ret receptor tyrosine kinase; g) compounds that target, reduce or inhibit Kit/SCFR receptor tyrosine kinase activity, such as imatinib; h) compounds that target, reduce or inhibit the activity of C-kit receptor tyrosine kinases that are part of the PDGFR family, such as compounds that target, reduce or inhibit the activity of members of the c-Kit receptor tyrosine kinase family, in particular compounds that inhibit c-Kit receptor, such as imatinib; i) compounds that target, reduce or inhibit the activity of c-Abl family members, their gene fusion products (e.g. BCR-Abl kinase) and mutant variants, such as compounds that target, reduce or inhibit the activity of c-Abl family members and products fusions of their genes, such as an N-phenyl-2-pyrimidinamine derivative such as imatinib or nilotinib (AMN107); PD180970; AG957; NSC 680410; PD173955 from ParkeDavis; or dasatinib (BMS-354825); j) compounds that target, reduce or inhibit the activity of members of the protein kinase C (PKC) family and the Raf family of serine/threonine kinases, members of the MEK, SRC, JAK/pan-JAK, FAK, PDK1, PKB/Akt, Ras/MAPK, PI3K families , SYK, TYK2, BTK and TEC and/or members of the cyclin-dependent kinase (CDK) family, including staurosporine derivatives such as midostaurin; examples of other compounds include UCN-01, safingol, BAY 43-9006, bryostatin 1, perifosine; ilmofosine; RO 318220 and RO 320432; GO 6976; lsis 3521; LY333531/LY379196; isoquinoline compounds; FTI; PD184352 or QAN697 (PI3K inhibitor) or AT7519 (CDK inhibitor); k) compounds that target, reduce or inhibit the activity of protein tyrosine kinase inhibitors, such as compounds that target, reduce or inhibit the activity of protein tyrosine kinase inhibitors, including imatinib mesylate (Gleevec™) or tyrphostin, such as Tyrphostin A23/RG-50810; AG 99; Tyrphostin AG 213; Tyrphostin AG 1748; Tyrphostin AG 490; Tyrphostin B44; (+) enantiomer of Tyrphostin B44; Tyrphostin AG 555; AG 494; Tyrphostin AG 556, AG957 and adaphostin, adamantyl ester (4-{[(2,5-dihydroxyphenyl)methyl]amino}benzoic acid; NSC 680410, adaphostin); 1) compounds that target, reduce or inhibit the activity of members of the epidermal growth factor receptor tyrosine kinase family (EGFR1 ErbB2, ErbB3, ErbB4 as homo- or heterodimers) and mutant variants thereof, such as compounds that target, reduce or inhibit the activity of members the epidermal growth factor (EGF) receptor family, in particular, compounds, proteins or antibodies that inhibit members of the EGF receptor tyrosine kinase family, such as EGF receptor, ErbB2, ErbB3 and ErbB4, or bind to EGF or EGF-related ligands, CP 358774, ZD 1839, ZM 105180.; trastuzumab (Herceptin™), cetuximab (Erbitux™), Iressa, Tarceva, OSI-774, Cl-1033, EKB-569, GW-2016, E1.1, E2.4, E2.5, E6.2, E6. 4, E2.11, E6.3 or E7.6.3 and 7H-pyrrolo[2,3D]pyrimidine derivatives; m) compounds that target, reduce or inhibit the activity of the c-Met receptor, such as compounds that target, reduce or inhibit the activity of c-Met, in particular compounds that inhibit the kinase activity of the c-Met receptor or antibodies that target the extracellular domain c-Met or HGF binding, n) compounds that target, reduce or inhibit the kinase activity of one or more members of the JAK family (JAK1/JAK2/JAK3/TYK2 and/or pan-JAK), including, but not limited to, PRT- 062070, SB-1578, baricitinib, pacritinib, momelotinib, VX-509, AZD-1480, TG-101348,

- 41 046788 tofacitinib and ruxolitinib; o) compounds aimed at reducing or inhibiting the kinase activity of PI3 kinase (PI3K), including, but not limited to, ATU-027, SF-1126, DS-7423, PBI-05204, GSK-2126458, ZSTK-474, buparlisib, pictrelisib, PF-4691502, BYL-719, dactolisib, XL-147, XL-765 and idelalisib; And; and q) compounds that target, reduce or inhibit the signaling effects of hedgehog (Hh) or smoothed receptor (SMO) protein pathways, including, but not limited to, cyclopamine, vismodegib, itraconazole, erizmodegib and IPI-926 (saridegib).

The term PI3K inhibitor as used herein includes, but is not limited to, compounds having inhibitory activity against one or more enzymes of the phosphatidylinositol 3-kinase family, including, but not limited to, PI3Ka, PI3Ky, PI3K<5„ PI3Ke, PI3KC2a , PI3K-C2e, PI3K-C2y, Vps34, p110-a, pHG-β, p110-y, p110-5, p85-a, p85-b, p55-y, p150, p101 and p87. Examples of PI3K inhibitors that can be used in the present invention include, but are not limited to, ATU-027, SF-1126, DS-7423, PBI-05204, GSK-2126458, ZSTK-474, buparlisib, picterlisib, PF-4691502, BYL-719, dactolisib, XL-147, XL-765 and idelalisib.

The term Bcl-2 inhibitor as used herein includes, but is not limited to, compounds having inhibitory activity against B-cell lymphoma protein 2 (Bcl-2), including, but not limited to, ABT-199, ABT-731, ABT -737, apogosipol, pan-Bcl-2 inhibitors Ascenta, curcumin (and its analogs), dual-acting Bcl-2/Bcl-xL inhibitors (Infinity Pharmaceuticals/Novartis Pharmaceuticals), Genasense (G3139), HA14-1 (and its analogs; see WO 2008118802), navitoclax (and its analogs, see US 7390799), NH-1 (Shenayng Pharmaceutical University), obatoclax (and its analogs, see WO 2004106328), S-001 (Gloria Pharmaceuticals), compounds TW series (Univ. of Michigan) and venetoclax. In some embodiments, the Bcl-2 inhibitor is a small molecule drug. In some embodiments, the Bcl-2 inhibitor is a peptidomimetic. The term BTK inhibitor as used herein includes, but is not limited to, compounds having Bruton's tyrosine kinase (BTK) inhibitory activity, including, but not limited to, AVL-292 and ibrutinib.

The term SYK inhibitor as used herein includes, but is not limited to, compounds having spleen tyrosine kinase (SYK) inhibitory activity, including, but not limited to, PRT-062070, R-343, R-333, Excellair, PRT-062607 and fostamatinib. Other examples of BTK inhibitory compounds and conditions that can be treated with such compounds in combination with the compounds of the present invention can be found in WO 2008039218 and WO 2011090760, the entire contents of which are incorporated herein by reference.

Other examples of SYK inhibitory compounds and conditions that can be treated with such compounds in combination with the compounds of the present invention can be found in WO 2003063794, WO 2005007623 and WO 2006078846, the entire contents of which are incorporated herein by reference.

Other examples of PI3K inhibitory compounds and conditions that can be treated with such compounds in combination with compounds of the present invention can be found in WO 2004019973, WO 2004089925, WO 2007016176, US 8138347, WO 2002088112, WO 2007084786, WO 20071 29161, WO 2006122806, WO 2005113554 and WO 2007044729, the entire contents of these references are incorporated herein by reference.

Further examples of JAK inhibitory compounds and conditions that can be treated with such compounds in combination with compounds of the present invention can be found in WO 2009114512, WO 2008109943, WO 2007053452, WO 2000142246 and WO 2007070514, the entire contents of which are incorporated herein by reference. .

Other antitumor compounds include compounds whose activity is mediated by a mechanism other than protein kinase or lipid kinase inhibition, such as thalidomide (Thalomid™) and TNP-470.

Examples of proteasome inhibitors suitable for use in combination with the compounds of the present invention include, but are not limited to, bortezomib, disulfiram, epigallocatechin-3-gallate (EGCG), salinosporamide A, carfilzomib, ONX-0912, CEP-18770 and MLN9708.

Compounds that target, reduce or inhibit the activity of protein phosphatase or lipid phosphatase are, for example, phosphatase 1, phosphatase 2A or CDC25 inhibitors, such as okadaic acid or a derivative thereof.

Compounds that induce cell differentiation processes include, but are not limited to, retinoic acid, α-γ- or δ-tocopherol, or α-γ- or δ-tocotrienol.

The term cyclooxygenase inhibitor as used herein includes, but is not limited to, Cox-2 inhibitors, 5-alkyl-2-arylaminophenylacetic acid and derivatives thereof, such as celecoxib (Celebrex™), etoricoxib, valdecoxib or 5-alkyl-2-arylaminophenylacetic acid, such as 5-methyl-2-(2'-chloro-6'-fluoroanilino)phenylacetic acid, lumiracoxib.

The term bisphosphonates as used herein includes, but is not limited to, etridone

- 42 046788 vuyu, clodronic, tiludronic, pamidronic, alendronic, ibandronic, risedronic and zoledronic acids. Etridonic acid is marketed under the trade name Didronel™. Clodronic acid is marketed under the trade name Bonefos™. Tiludronic acid is marketed under the trade name Skelid™. Pamidronic acid is marketed under the trade name Aredia™. Alendronic acid is marketed under the trade name Fosamax™. Ibandronic acid is marketed under the trade name Bondranat™. Risedronic acid is marketed under the trade name Actonel™. Zoledronic acid is marketed under the trade name Zometa™. The term mTOR inhibitors refers to compounds that inhibit mammalian target of rapamycin (mTOR) and which have antiproliferative activity, such as sirolimus (Rapamune®), everolimus (Certican™), CCI-779 and ABT578.

The term heparanase inhibitor as used herein refers to compounds that target, reduce or inhibit the degradation of heparin sulfate. The term includes, but is not limited to, PI-88. The term biological response modifier in the present invention refers to lymphokine or interferons.

The term inhibitor of oncogenic Ras isoforms, such as H-Ras, K-Ras or N-Ras, as used herein refers to compounds that target, reduce or inhibit the oncogenic activity of Ras; an example is a farnesyltransferase inhibitor such as L-744832, DK8G557 or R115777 (Zarnestra™). As used herein, the term telomerase inhibitor refers to compounds that target, reduce or inhibit telomerase activity. Compounds that target, reduce or inhibit telomerase activity are, in particular, compounds that inhibit the telomerase receptor, such as telomestatin.

The term methionine aminopeptidase inhibitor as used herein refers to compounds that target, reduce or inhibit the activity of methionine aminopeptidase. Compounds that target, reduce or inhibit methionine aminopeptidase activity include, but are not limited to, benhamide or a derivative thereof.

The term proteasome inhibitor as used herein refers to compounds that target, reduce or inhibit proteasome activity. Compounds that target, reduce or inhibit proteasome activity include, but are not limited to, Bortezomib (Velcade™) and MLN341.

The term matrix metalloproteinase inhibitor or (MMP inhibitor) as used herein includes, but is not limited to, peptidomimetic and non-peptidomimetic collagen inhibitors, tetracycline derivatives, such as the hydroxamate peptidomimetic inhibitor batimastat and its orally bioavailable analog, marimastat (BB). -2516), prinomastat (AG3340), metastat (NSC 683551), BMS-279251, BAY 12-9566, TAA211, MMI270B or AAJ996. The term compounds used in the treatment of hematological malignancies in the present invention includes, but is not limited to, FMS-like tyrosine kinase inhibitors, which are compounds that target, reduce or inhibit the activity of FMS-like tyrosine kinase receptors (Flt-3R); interferon, 1-β-Οarabinofuransylcytosine (ara-c) and bisulfan; and ALK inhibitors, which are compounds that target, reduce or inhibit anaplastic lymphoma kinase activity. Compounds that target, reduce or inhibit the activity of FMS-like tyrosine kinase receptors (Flt-3R) are, in particular, compounds, proteins or antibodies that inhibit members of the Flt-3R receptor kinase family, such as PKC412, midostaurin, a derivative staurosporine, SU11248 and MLN518.

The term HSP90 inhibitors as used herein includes, but is not limited to, compounds that target, reduce or inhibit the ATPase activity inherent in HSP90; involved in the degradation of, targeting, reducing, or inhibiting HSP90-dependent proteins through the ubiquitinin-proteasome pathway. Compounds that target, reduce or inhibit the ATPase activity inherent in HSP90 are, in particular, compounds, proteins or antibodies that inhibit the ATPase activity of HSP90, such as 17allylamino, 17-demethoxygeldanamycin (17AAG), a derivative of geldanamycin; other geldanamycin-related compounds; radicicol and HDAC inhibitors. The term antiproliferative antibodies as used herein includes, but is not limited to, trastuzumab (Herceptin™), TrastuzumabDM1, Erbitux, bevacizumab (Avastin™), rituximab (Rituxan®), PRO64553 (anti-CD40) and 2C4. Antibodies include intact monoclonal antibodies, polyclonal antibodies, polyspecific antibodies derived from at least 2 intact antibodies, and antibody fragments, provided they exhibit the desired biological activity.

For the treatment of acute myeloid leukemia (AML), the compounds of the present invention can be used in combination with standard treatments for leukemia, in particular in combination with treatments used for the treatment of AML. In particular, connections according to this

- 43 046788 the invention can be administered in combination with, for example, farnesyltransferase inhibitors and/or other drugs suitable for use in the treatment of AML, such as daunorubicin, adriamycin, Ara-C, VP-16, teniposide, mitoxantrone, idarubicin, carboplatin and PKC412. Other antileukemic compounds include, for example, Ara-C, a pyrimidine analogue, which is a 2'-alpha-hydroxyribose (arabinoside) derivative of deoxycytidine. Also included are the purine analogue of hypoxanthine, 6-mercaptopurine (6-MP), and fludarabine phosphate. Compounds that target, reduce, or inhibit histone deacetylase (HDAC) activity, such as sodium butyrate and suberoylanilide hydroxamic acid (SAHA), inhibit the activity of enzymes known as histone deacetylases. Specific HDAC inhibitors include MS275, SAHA, FK228 (formerly FR901228), trichostatin A and the compounds described in US 6552065, including, but not limited to, Nahydroxy-3-[4-[[[2-(2-methyl-1H -indol-3-yl)ethyl]amino]methyl]phenyl]-2E-2propenamide or a pharmaceutically acceptable salt thereof and H-hydroxy-3-[4-[[(2-hydroxyethyl)[2-(1Nindol-3-yl) )ethyl]amino]methyl]phenyl]-2E-2-propenamide or a pharmaceutically acceptable salt thereof, in particular the lactate salt. In the present invention, somatostatin receptor antagonists refer to compounds that target, act on or inhibit the somatostatin receptor, such as octreotide and SOM230. Approaches based on tumor cell damage include approaches such as ionizing radiation. The term ionizing radiation, as referred to above and below, means ionizing radiation that occurs in the form of either electromagnetic radiation (such as X-rays and gamma rays) or particles (such as alpha and beta particles). Ionizing radiation has been proposed for, without limitation, radiation therapy and is known in the art. See Hellman, Principles of Radiation Therapy, Cancer, in Principles and Practice of Oncology, Devita et al., Eds., 4th Edition, Vol. 1, pp. 248-275(1993).

Also included are EDG binders and ribonucleotide reductase inhibitors. The term EDG binders as used herein refers to a class of immunosuppressants that modulate lymphocyte recycling, such as FTY720. The term ribonucleotide reductase inhibitors refers to pyrimidine or purine nucleoside analogues, including, but not limited to, fludarabine and/or cytosine arabinoside (ara-C), 6-thioguanine, 5-fluorouracil, cladribine, 6-mercaptopurine (particularly in combination with ara -C against ALL) and/or pentostatin. Ribonucleotide reductase inhibitors are, in particular, hydroxyurea or 2hydroxy-1H-isoindol-1,3-dione derivatives.

Also included, but not limited to, are anti-vascular endothelial growth factor (VEGF) compounds, proteins or monoclonal antibodies such as 1-(4-chloroanilino)-4-(4-pyridylmethyl)phthalazine or its pharmaceutically acceptable salt, 1-(4) succinate -chloranilino)-4-(4-pyridylmethyl)phthalazine; Angiostatin™; Endostatin™; anthranilic acid amides; ZD4190; ZD6474; SU5416; SU6668; bevacizumab; or anti-VEGF antibodies or anti-VEGF receptor antibodies such as rhuMAb and RHUFab, a VEGF aptamer such as Macugon; FLT-4 inhibitors, FLT-3 inhibitors, anti-VEGFR-2 IgG1 antibody, angiozyme (RPI 4610) and bevacizumab (Avastin™).

Photodynamic therapy in the present invention refers to a therapy in which certain chemicals known as photosensitizing compounds are used to treat or prevent cancer. Examples of photodynamic therapy include treatment with compounds such as Visudyne™ and sodium porfimer. Angiostatic steroids in the present invention refer to compounds that block or inhibit angiogenesis, such as, for example, anecortave, triamcinolone, hydrocortisone, 11a-epihydrocortisol, cortexolone, 17a-hydroxyprogesterone, corticosterone, deoxycorticosterone, testosterone, estrone and dexamethasone.

Implants containing corticosteroids include compounds such as fluocinolone and dexamethasone.

Other chemotherapeutic compounds include, but are not limited to, plant alkaloids, hormonal compounds and antagonists; biological response modifiers, preferably lymphokines or interferons; antisense oligonucleotides or derivatives of oligonucleotides; shRNA or siRNA; or other compounds or compounds with a different or unknown mechanism of action.

The structure of active compounds was identified by code numbers, nonproprietary names or trademarks, which can be obtained from the current edition of the standard manual The Merck Index or from databases such as Patents International (eg, IMS World Publications).

The compound of the present invention can also be used in combination with known therapeutic processes, for example the administration of hormones or radiation. In some embodiments, the proposed compound is used as a radiosensitizer, in particular for the treatment of tumors that exhibit poor sensitivity to radiotherapy. A compound of the present invention may be administered alone or in combination with one or more other therapeutic compounds, with possible combination therapy taking the form

- 44 046788 fixed combinations or administration of a compound according to the present invention and one or more other therapeutic compounds at intervals of time independently of each other or combined administration of fixed combinations and one or more other therapeutic compounds. The compound of the present invention may be administered as a substitute or additive, particularly for the treatment of a tumor in combination with chemotherapy, radiotherapy, immunotherapy, phototherapy, surgery, or a combination thereof. Long-term therapy is also possible, an example being adjuvant therapy in the context of other treatment strategies as described above. Another possible treatment is therapy to maintain the patient's condition after tumor regression or chemoprophylactic therapy, for example in patients at risk. These additional agents can be administered separately from the proposed composition X4P-001, as part of a multiple-dose regimen. Alternatively, these agents may be part of a single dosage form when mixed with the proposed composition X4P-001 to form a single composition. When administered as part of a multiple-dose regimen, the two active agents may be administered simultaneously, sequentially, or at intervals typically within 5 hours.

As used herein, the terms combination, combination and related terms refer to the simultaneous or sequential administration of therapeutic agents in accordance with the present invention. For example, a compound of the present invention may be administered with another therapeutic agent simultaneously or sequentially in a divided dose dosage form or together in a single dosage form. Accordingly, the present invention provides a single dosage form containing a compound of the present invention, an additional therapeutic agent and a pharmaceutically acceptable carrier, adjuvant or vehicle. The amount of the proposed composition X4P-001 and additional therapeutic agent (in compositions that contain the additional therapeutic agent described above) that can be combined with carrier materials to form a single dosage form may vary depending on the host being treated and the specific route introduction. Preferably, the compositions of the present invention should be formulated such that a dose of 0.01-100 mg/kg body weight/day of a compound of the present invention can be administered. In compositions containing an additional therapeutic agent, the additional therapeutic agent and the compound of the present invention may act synergistically. Thus, the amount of additional therapeutic agent in such compositions will be less than that required for monotherapy using only that therapeutic agent. In such compositions, a dose of 0.01 to 1000 μg/kg body weight/day of the additional therapeutic agent can be administered. The amount of additional therapeutic agent present in the composition of the present invention will be no more than would normally be administered in a composition containing said therapeutic agent as the sole active agent. Preferably, the amount of additional therapeutic agent in the proposed X4P-001 compositions will be from about 50 to 100% of the amount typically present in a composition containing such agent when it is the only therapeutically active agent. The compounds of the present invention or pharmaceutical compositions thereof may also be included in compositions for coating implantable medical devices such as prostheses, prosthetic valves, vascular grafts, stents and catheters. Vascular stents, for example, are used to avoid restenosis (re-narrowing of the vessel wall after injury). However, patients with stents or other implantable devices are at risk for clot formation or platelet activation. Such undesirable effects can be prevented or reduced by precoating the device with a pharmaceutically acceptable composition containing a kinase inhibitor. Implantable devices coated with the compound of the present invention represent another embodiment of the present invention.

For a more complete understanding of the invention described in the present invention, the following examples are provided. It should be understood that these examples are provided for illustrative purposes only and are not intended to limit the invention in any way.

Examples

As shown in the examples below, in some illustrative embodiments, compounds are prepared in accordance with the following general methods. It should be understood that while the general methods illustrate the synthesis of some of the compounds of the present invention, the following general methods and other methods known to one skilled in the art can be applied to all compounds and the subclasses and species of each of these compounds described in the present invention.

Preparation methods applicable to some of the compounds of the present invention are described in Crawford et al. (2008) Org. Process Res. Dev. 12:823-830; US patent No. 7354934, sources WO 00/56729, USSN 60/232891 and USSN 60/234510, and An, H.; Wang, T.; Mohan, V.; Griffey R. H.; Cook, P. D. Tetrahedron 1998, 54, 3999-4012; the contents of each of which I will fully include

- 45 046788 incorporated into this document by reference. One skilled in the art will be able to modify such described methods, using only routine experimentation, to provide alternative routes for preparing, testing and analyzing the compounds of the present invention.

Example 1: Methylimine Impurity

High performance liquid chromatography-mass spectrometry (HPLC-MS) analysis indicated that the peak with a relative retention time (RRT) of 1.13 corresponded to [M+1]+ having m/z 362 (12 mass units more than X4P-001 ). This peak disappeared after several hours if X4P-001 was solubilized in an acidic aqueous medium. The molecular weight of the impurity, in combination with its chemical properties, indicated a methylimine structure formed due to the conversion of the primary amine. Without being bound by any particular theory, the present inventors believe that Compound I-1 was formed in the presence of formaldehyde or a source of formaldehyde. The imine I-1 impurity and the likely route of formation are shown in Scheme 1 below.

Scheme 1

Example 2: N-formyl impurity

According to HPLC-MS analysis, the peak with an RRT of 1.28 corresponded to [M+1] ⁺ having m/z 378 (28 mass units more than X4P-001), indicating an N-formyl derivative of X4P-001 . This was confirmed by independent synthesis. Direct reaction of X4P-001 with ethyl formate resulted in the formation of N-formyl compound 1-2, which was identical to the impurity in the tested batch based on MS and HPLC data. The formation of formate is shown in Scheme 2 below.

Scheme 2

Example 3: Aldehyde Impurity

The peak with an RRT of 1.14 was characterized as I-6. Without being bound by any particular theory, the present inventors believe that I-6 was formed by oxidation of the amino group in the alkyl chain of X4P-001, as shown in Scheme 3 below.

Scheme 3

Example 4: Acetamide Impurity

Based on HPLC-MS analysis, the peak with an RRT of 1.39 corresponded to [M+1] ⁺ having m/z 392 (42 mass units greater than X4P-001), which was characterized as acetamide X4P-001. It was formed at elevated temperatures by the reaction of X4P-001 with the crystallization solvent isopropyl acetate, as shown in Scheme 4. Independent synthesis of the acetamide impurity confirmed this structure. Reaction of X4P-001 with acetic anhydride produced acetamide I-5, which was identical to the impurity in the batch tested based on MS and HPLC data. Isopropyl acetate is used in some methods for preparing X4P-001 in free base form, which involve the use of X4P-001 in the form of a salt with n-hydroxybenzoic acid.

- 46 046788

Scheme 4

Example 5: Benzimidazole Impurity

Based on LC-MS analysis, the peak with an RRT of 1.67 corresponded to [M+1]+ having m/z 481 (131 mass units greater than X4P-001), which was characterized as the M ⁺ benzimidazole compound I- 3. It was formed as an impurity during the protection reaction of 2chloromethylbenzimidazole with tert-butoxycarbonyl (Boc group), as shown in Scheme 5 below. During the N-alkylation reaction (step 2 of the X4P-001 synthesis), the impurity was alkylated and, as a result, was present throughout the remainder of the preparation process.

Scheme 5

( ²⁸⁹ °)Admixture

1. 2894 (alkylation)

2. Removal of the protecting group

3. Crystallization ► Х4Р001

Independent synthesis involving a secondary amine reaction

2918 with Boc-2chloromethylbenzimidazole 2890 (I-9) followed by deprotection resulted in I-3, which was identical to the observed impurity based on MS and HPLC data.

Example 6 Aminal Impurity

The aminal impurity arose during the preparation of X4P-001 in the form of a salt with n-hydroxybenzoic acid as a result of the reaction of the imine impurity with n-hydroxybenzoic acid. This impurity relates to processes for the preparation of X4P-001 that use X4P-001 in the form of a salt with n-hydroxybenzoic acid or intermediates thereof.

The structure of aminal impurity I-4 and its origin are shown below in Scheme 6.

Scheme 6

Example 7: Basis for establishing acceptance criteria for organic impurities

The results of determining the content of impurities for the corresponding batches of X4P-001 and its salt with n-hydroxybenzoic acid (PHBA) are presented below in table. 2. Lots of X4P-001 taken for initial toxicology studies were first used to develop impurity specifications. Batches of X4P-001 n-hydroxybenzoic acid salt were used in long-term toxicology studies and are also presented in Table 1. 2. The total amount of impurities in these six lots ranged from <0.05 to 1.4%, with individual unidentified impurities of >0.05% present in only a single lot at 0.1%. Taking into account the limited number of batches received and the impurity profiles of the batches of X4P-001 used for toxicological and clinical studies, the total impurity content in the drug substance release specification was assumed to be <3.0% w/w, with unidentified impurity from a single batch should not exceed 0.2% w/w (0.5% for stability) (Table 2). Content of all identified individual impurities and n-hydroxybenzoic

- 47 046788 acid was required to be <0.5% w/w, with the exception of the imine impurity, which was assumed to be 1.1% based on the amount of this impurity administered in toxicological studies. In a separate batch of product (not shown in Table 2), namely batch 3-1 X4P-001 (Table 4), a high purity drug substance was provided with a total impurity content of 1.20% w/w, with In this case, the content of none of the identified impurities exceeded 0.07% w/w, with the exception of the imine impurity, which amounted to 0.62% w/w. The result of the analysis of the batch of drug substance X4P-001 for clinical use was 99, 3% w/w, and chiral purity was >99% enantiomeric excess (E.I.).

table 2

Batch number 2-1 2-2 2-3 2-4 2-5 2-6 2-7 Type of drug substance Free base Free base Free base Free base Free base Salt PHB K Salt PGBC Individual impurities according to HPLC data (% w/w) in terms of anhydrous substance Research results Imin (1-1) 1.1 0.5 0.1 0.49 0.65 0.02 0.02 Formil (1-2) 0.2 0.1 0.1 0.13 0.13 BUT. BUT. Acetamide (1-5) 0.1 0.1 0.1 0.10 0.06 - - Aminal (1-4) - - - - - 0.01 But. Benzimidazole (1-3) 0.01 ^a WELL. 0.2 <pko 0.11 0.01 0.01 Unidentified in a single batch WELL. WELL 0.1 But. WELL But. But. Contents Ш БК — — — — — 28.1 28.4 Total impurity content (% w/w) ^b 1.4 0.7 0.5 0.72 1.01 < 0.05 <0.05 % e.i. (R)enantiomer according to HPLC data >99 >99 >99 >99 >99 >99 >99

Abbreviations used:

^a Value below LOQ (limit of quantitation) ^b Specification applies only to the drug substance for clinical use in free base form obtained by using X4P-001 in salt form with n-hydroxybenzoic acid as the starting material -=Not applicable

N.O.=Undefined

N.M.=No less

N.B.=No more

N.U.=Not specified

Typically, the wt.% of each impurity is determined by HPLC, and measurements are made either initially or after storage, and optionally on an ongoing basis during the shelf life of the X4P-001 composition. In some embodiments, the impurity content is determined after storing the composition under test conditions, which are conditions of elevated temperature, humidity, or both, used to simulate the effect of long-term storage under ambient conditions. Accordingly, in some embodiments of the present invention there is provided a composition X4P-001 containing no more than 1.1% imine (I-1); no more than 0.3% formyl (I-2); no more than 0.4% benzimidazole (I-3); no more than 0.5% aminal (I-4); no more than 0.5% acetamide (I-5); no more than 0.4% aldehyde (I-6); no more than 0.3% NT-316 without Boc group (I-7) and no more than 0.2% unidentified impurity from a single batch. In addition, when a drug substance for clinical use in free base form is prepared using X4P-001 in n-hydroxybenzoic acid salt form, PHBA is present in the composition in an amount of not more than 1.0%. In some embodiments, the total amount of impurities present is no more than 3.0%. In some embodiments, the enantiomeric excess (% ee) of the R-enantiomer is at least 97.0%.

Example 8: Identification of Impurities Formed After Storage

Stability analysis by MET/CR/1448 of samples of X4P-001 (PTL/ST/0511) stored at 25°C/60% RH (25/60) for three months showed the formation of two unknown impurities.

Unknown impurity 1 - RRT 1.14

- 48 046788

Unknown impurity 2 - RRT 1.24

The impurity with RRT 1.14 was also the main degradation product in PTL/DA/0175 under elevated temperature and humidity conditions (80°C/80% RH).

Brief information

Two impurities formed during the stability study of X4P-001 over a period of t=3 months at 25°C/60% RH and during the forced degradation study at elevated temperature/humidity were identified by LC-MS.

Impurity with RRT 1.14 MET/CR1448 Aldehyde - oxidation product Empirical formula = C21H24N4O Monoisotopic mass = 348.195

Admixture with RRT 1.24 MET/CR1448

Acetamide impurity Empirical formula = C23H29N5O Monoisotopic mass = 391.237

experimental part

Device parameters

The work was performed in accordance with Good Manufacturing Practice (GMP) requirements on a Waters Alliance HPLC system coupled to a ZQ 2000 single quadrupole mass spectrometer. Data were collected using Empower 2 software. Impurity Analysis and Determination Method for X4P-001 (MET /CR/1448) is not ideal for mass spectrometry because it involves the use of TFA as a modifier in the eluent, which can significantly suppress ionization. Initially, TFA was used instead of 0.15% v/v. formic acid. The flow rate was also changed to allow direct connection to the mass spectrometer (maximum flow rate 0.3 ml/min) and the gradient was set accordingly.

HPLC conditions 1:

Column: Zorbax Bonus-RP, 150x4.6 mm, 3.5 µm

Sample injection volume: 100 µl

Detection: UV at 220 nm (190-400 nm)

MS ER+100-700 Yes, ER - 100-700 Yes

Mobile phase A: 0.15% formic acid in water

Mobile phase B: 0.15% formic acid in MeCN

Time(min) % A %IN 0.0 95 5 6 95 5 51 5 95 60 5 95 60.1 95 5 75 95 5

Flow rate: 0.3ml/min

Column temperature: ambient temperature

Analysis run time: 75 min

MS 1 settings

- 49 046788

Ion Source Tab ER + ER- Capillary voltage (kV) 3.5 4 Cone voltage (V) 25 25 Extractor voltage (V) 3 0 RF Lens 0.4 3 Source temperature (°C) 150 150 Desolvation temperature (°C) 150 150 Drying gas flow rate (l/h) 300 300 Gas flow rate in cone (l/h) 50 50 Analyzer tab Resolution in the low molecular weight range 15 15 Resolution in the high molecular weight range 15 15 Ionization energy 1.8 2 Multiplier 521 521

X4P-001 was not retained under these conditions because it required ion pairing with TFA. The resulting chromatograms and mass spectra are presented in Fig. 5.

Estimated masses for two impurities were obtained. All mass spectra presented were background corrected (the region taken immediately before and after the target peak).

Unknown impurity 1 MET/CR/1448 RRT 1.14 [Μ+Η]^/ζ 349.2

Unknown impurity 2 MET/CR/1448 RRT 1.24 [Μ+Η]^/ζ 392.3

The present inventors hypothesized that the impurities in question were likely co-eluting with other impurities. Accordingly, the samples were analyzed using chromatographic conditions according to MET/CR/1448. Concentrations and volumes of sample injections were increased to counter TFA-induced ionization suppression.

HPLC Conditions 2:

Column: Zorbax SB-C8, 150 mm x 4.6 mm, 3.5 µm

Pre-column: Zorbax SB-C8, 12.5 mmx4.6 mm, 5 µm

Sample Injection Volume: Varies

Detection: UV at 270 nm

Mobile phase:

Mobile phase A: 0.2% TFA in water

Mobile phase B: 0.1% TFA in acetonitrile

Gradient elution

Time (min) % A %IN Flow rate(ml/min) 0 92 8 0.8 5 90 10 0.8 15 89 AND 0.8 25 80 20 0.8 28 80 20 0.8 37 55 45 0.8 44 20 80 0.8 47 20 80 0.8 48 92 8 1.2 53 92 8 1.2 54 92 8 0.8 55 92 8 0.8

Initial flow rate: 0.8 mL/min (relative to gradient program), 4:1 split at mass spectrometer inlet. Column temperature: 25°C

Analysis run time: 55 min

MS parameters corresponded to Settings 1 above. Various X4P-001 samples in which the impurities in question were detected were prepared in methanol at a concentration of 1 or 10 mg/ml:

PTL/ST/0511, batch 3-1, 25/60, t=3 months

- 50 046788

PTL/DA/0175, sample for degradation studies at 80°C/80% RH (80/80), t=1 and 7 days.

The obtained data are presented in Fig. 6-9.

Mass spectrometric data obtained under HPLC conditions 2 were confirmed:

Unknown impurity 1 MET/CR/1448 RRT 1.14 [M+H]=m/z 349.2

Unknown impurity 2 MET/CR/1448 RRT 1.24 [M+H]=m/z 392.3

Finally, the chromatography conditions were modified to allow injection into the mass spectrometer without splitting the flow. Results for HPLC conditions 3 are shown in FIG. 10.

HPLC Conditions 3:

Column: Zorbax SB-C8, 150 mm x 4.6 mm, 3.5 µm

Pre-column: Zorbax SB-C8, 12.5 mmx4.6 mm, 5 µm

Sample Injection Volume: Varies

Detection: UV at 270 nm

Mobile phase: Mobile phase A: 0.2% TFA in water

Mobile phase B: 0.1% TFA in acetonitrile __________Gradient elution

Time (min) % A % IN 0 92 8 13.3 90 10 39.9 89 AND 66.50 80 20 74.48 80 20 98.42 55 45 117 20 80 125 20 80 125.10 92 8 140 92 8

Flow rate: 0.3 ml/min, direct injection into mass spectrometer

Column temperature: 25°C

Analysis run time: 140 min

The MS parameters corresponded to Settings 1 above.

The discussion of the results

All obtained spectra confirmed the original results, even though the signals and resolution of the X4P-001 peaks varied.

Unknown impurity 1 MET/CR/1448 RRT 1.14 [Μ+Η]^ζ 349.2

Unknown impurity 2 MET/CR/1448 RRT 1.24 [Μ+Η]^ζ 392.3

Unknown impurity 1 with RRT 1.14 (aldehyde)

A molecular weight of 348 indicates the loss of a nitrogen atom (nitrogen rule). This corresponds to the oxidation of the amino group in the alkyl chain to form an aldehyde. The proposed structure is shown below.

Empirical formula^^^^

Monoisotopic mass=348.195

Unknown impurity 2 with RRT 1.24 (acetamide)

Molecular weight 391 corresponds to I-5, an acetamide impurity (formed by the reaction of X4P-001 with residual isopropyl acetate). The structure of I-5 is shown below.

- 51 046788

1-5

Empirical formula = C23H ₂₉ K ₅ O

Monoisotopic mass=391.237

conclusions

Both unknown impurities were identified from the obtained mass spectrometry data. The impurity with RRT 1.14 was an oxidation product of aldehyde, and the impurity with RRT 1.24 was an acetamide I-5 impurity.

Example 9. Improvements to the method for producing X4P-001 to reduce and control the content of impurities

Introduction

The final stage of the previous method for obtaining a drug substance in accordance with GMP to support clinical studies (method option 2; shown in Fig. 2) began with the use of compounds NT-316 and AMD-2890 (I-9). After studying the structure and origin of the most significant impurities formed during this preparation process, a new method (method 3 option) was developed to provide more effective control of the impurity profile as well as API crystallization. Moreover, this work led to significant improvements in the reproducibility and reliability of the final stage of the method.

Description of the synthesis route and key intermediates of API X4P-001 was obtained in 2 stages by convergent synthesis starting from two key fragments: NT-316 and AMD-2890. The penultimate intermediate AMD-11070 was not isolated but was immediately converted to the API, which was crystallized after the workup procedure.

Scheme 7 and Scheme 8 show the synthesis of the X4P-001 API molecule.

Scheme 7: Initial stages of the synthesis of X4P-001

Scheme 8: Final stages of the synthesis of X4P-001

Х4Р-001

The key fragment AMD-2890 was synthesized in one step from RM-396 (I-8). Although RM-396 is commercially available, its purity varies greatly depending on its source. The purity of the commercially available substance may appear to be high (>97% peak area) by HPLC analysis, but the true purity in wt% was found to be only 90%. It is likely that RM-396 undergoes self-alkylation, resulting in the formation of a variable mixture of oligomeric impurities. Only after converting RM-396 to AMD-2890 was a stable compound with a well-defined impurity profile obtained.

- 52 046788

Another key fragment is NT-316, a stable, crystalline and well-characterized compound that comprises a significant portion of the API structure. NT-316 was prepared by convergent synthesis via the reaction of NT-319 with the chiral amine salt NT-272. NT-272 is a stable crystalline compound that can be provided through custom synthesis.

NT-319 is an extremely unstable intermediate that cannot be isolated without degradation. It was prepared in 3 steps (without isolating intermediates in each step) from commercially available ABA. Monitoring the purity of ABA is difficult, not only because it is an oil, but also because it lacks a UV chromophore, which severely limits the methods suitable for its accurate analytical testing.

Based on the above, it is the impurity profile of these two fragments, NT-316 and AMD2890 (I-9), combined with the precisely defined conditions of the two final chemical synthesis steps (which should be conducted under GMP compliance controls) that determine the impurity profile and quality of the API.

Thus, the chemical and enantiomeric purity of NT-316 and the chemical purity of AMD-2890 can be directly correlated with the chemical and enantiomeric purity of the API (X4P-001).

Description of key transformations in accordance with the method

In fig. Figure 3 shows the difference between the new method (method 3) and the previous version (method 2). In Method 2, it was first necessary to isolate the API as its 4-hydroxybenzoate salt. This salt is then converted in a separate step to the corresponding free base to produce the API. The chemical synthesis steps according to method variant 3 remained unchanged. Compounds NT-316 and AMD-2890 (I-9) were reacted with each other to produce the product (tri-Boc), which was not isolated but immediately deprotected to yield the crude API. However, the method used to isolate the API was significantly modified in a variant of Method 3.

The most significant impurities that are formed in the case of method option 2 are imine A (RRT 1.08) and N-formyl B (RRT 1.28), as shown in Scheme 9. These impurities were shown to be due to the use of dichloromethane in as a solvent in the extraction process of crude API. Additionally, the formation of acetamide C impurity (RRT 1.37) can be correlated with the use of isopropyl acetate as a solvent for the final crystallization of the API in the free base form.

Scheme 9: Impurities found in bulk drug substance

Thus, dichloromethane and isopropyl acetate, which were used in the preparation and isolation of X4P-001 according to option 2, were replaced by 1-butanol and a mixture of toluene/methanol, respectively, in option 3. The authors of the present invention found that these solvents did not react with API, and, accordingly, according to the authors, this replacement led to the significant decrease in the content of impurities A (imine), B (N-formyl) and C (acetamide) observed by them.

In addition, it was discovered that problems associated with gum formation and API oiling during final product crystallization in Process Option 2 were due to the quality of the AMD-2890 starting material. This substance is derived from commercially available 2chloromethylbenzimidazole (RM-396), which may have a low wt% purity even though the peak area% purity appears to be high (>98%). Development of an improved method

- 53 046788 recovery of AMD-2890 is an integral part of Process Option 3, and this has consistently resulted in high quality AMD-2890 that is not only excellent in % purity, but also in % w/w purity.

Further optimization of aeration and carbon treatment during the API isolation process resulted in better control of the color of the isolated API, and these operations are also included in Method 3 option.

Finally, the conditions for crystallization of the API in the free base form from toluene were strictly defined. The region of the metastable state and the optimal point of seeding were established. The present inventors have also determined the optimal cooling rate after seed layer formation and created a suitable protocol for washing and drying.

In fig. 4 provides a detailed comparison of method options 2 and 3 for sequential operations in obtaining and isolating the API.

The reliability and reproducibility of Method 3 was demonstrated by conducting three identical laboratory experiments on a 100 g scale. As shown in the data in Table. 3 and table. 4, the demonstration of this method was successful. The present inventors further scaled up this method to a quantity of 10 kg. A total of 9.75 kg of X4P-001 was produced in accordance with GMP in one batch with almost identical results. A detailed discussion of these experiments is provided below.

Description of a variant of method 3 and improvements to method 1 and method 2

The following section provides a brief description of the various steps involved in the synthesis and isolation of the API in the final step of Method 3.

Synthesis of NT-316

The initial steps of the process for producing the key starting material NT-316 from the custom-synthesized chiral amine NT-272 and ABA remained unchanged from previous production cycles.

Synthesis of AMD-2890

As noted above, the synthesis of AMD-2890 (I-9) was improved, particularly its isolation and crystallization, to account for the varying purities and colors of commercially available 2chloromethylbenzimidazole (RM-396). This substance is usually dark brown to black in color. Therefore, charcoal treatment was introduced to provide better color control to the AMD-2890.

Thus, 2-chloromethylbenzimidazole (RM-396) was reacted with 1.3 eq. di-tert-butyl dicarbonate in 8.6 volumes (vol.) of DMF (KN-dimethylformamide) at 40°C in the presence of 0.1 equivalent (eq.) DIPEA (diisopropylethylamine). After completion of the reaction, decolorizing charcoal was added. After 1.5 hours at 40°C, the mixture was filtered and the solids were washed with 1 volume of DMF. Water (3.5 vol) was slowly added to the filtrates to obtain a slightly cloudy mixture, to which 1% AMD-2890 was added as a seed. During aging, a suspension formed, which was left to slowly cool to 20°C. After slowly adding additional water (1 vol.) and further cooling to 0°C, the suspension was filtered. The solids were first washed with a 2:1 mixture of DMF and water, then with water (2x3 volumes), performing all washes at 0°C. The filter cake was dried under a stream of nitrogen to obtain light yellow AMD-2890. This compound typically had a purity of >99% peak area (HPLC) and >99 wt% purity (NMR).

Synthesis and isolation of X4P-001

As before, equimolar amounts of NT-316 and AMD-2890 were reacted in the presence of diisopropylethylamine (DIPEA) and tetrabutylammonium iodide (TBAI) in acetonitrile at 60-65°C. After completion of the reaction, the mixture was cooled to ambient temperature, and the reaction was quenched with 0.3 vol. ammonia, after which 1 vol. was added. water. Next, the resulting mixture was added to a mixture of 2 vol. concentrated hydrochloric acid and 3 vol. water. Incubation for several hours at 35-40°C in this acidic solution resulted in the removal of the protecting groups in the protected intermediate to form the API. The acetonitrile was then removed by vacuum distillation.

At this point, 2 volumes of 1-butanol were added and the pH was adjusted to 12 with 20% NaOH solution. The resulting two-phase mixture was bubbled with a mixture of 10% oxygen in nitrogen for 2 hours at 20°C. The pH was then adjusted to 3.0-3.5 using a solution of 18% HCl in water. The two resulting phases were separated and the aqueous layer was washed with 1-butanol (3x3 volume). The organic layers were combined and extracted with 3 volumes of water. All aqueous phases were then combined and 0.4 wt.% carbon was added. After 1-2 hours the mixture was filtered and the solids were washed with 3 volumes of water.

Toluene (7 volumes) and methanol (1 volume) were added to the combined filtrates and the temperature was increased to 45-55°C, after which the pH was adjusted to 9.5-10.0 using a 20% sodium hydroxide solution. The phases were separated and the aqueous layers were extracted two more times with 3 volumes of toluene.

All toluene layers were combined and partially concentrated by vacuum distillation at 45-50°C. After several additions of fresh toluene and continued vacuum distillation at 45-50°C to remove other volatile solvents, an API solution of approximately 3 volumes was obtained

- 54 046788 toluene. This solution was heated to 60°C and purified by in-line filtration.

After cooling to 50°C, up to 0.5 wt.% X4R-001 was carefully added to this solution as a seed. After the formation of the seed layer, equilibrium crystallization of the API was allowed for 2-3 hours, after which the suspension was left to slowly cool to 0°C. To stimulate crystal growth, the resulting suspension was gradually heated back to 30-35°C and then allowed to cool again to 0°C. Finally, the suspension was filtered and the resulting solids were washed with toluene and dried in a filter dryer by heating under vacuum at 60° C. for 16 hours with occasional gentle stirring.

The use of this technique was demonstrated on a 10 kg scale, resulting in X4P-001 with a purity of 99.0-99.5% peak area (>99.9% enantiopurity) and a residual toluene content of 1337 ppm

Comparison of method 2 and method 3

In table 3 shows a comparison of key process parameters and results for batches recently produced using Methods 2 and 3. As mentioned above, a variant of Method 3 was first demonstrated at laboratory scale and then scaled up in a plant to produce X4P-001. In table Figure 4 shows a comparison of the impurity profiles in these batches.

What is especially important, in table. Figure 4 shows that moving from Method Option 2 to Method 3 consistently resulted in a reduction in total impurities overall of 0.5-1.1% of peak area, as well as a significant reduction in total residual solvent content of the API without adversely affecting other important results provided by this method.

Table 3. Comparison of key parameters of the method and finished batches obtained using method options 2 and 3

Batch number 3-1 3-2 3-3 3-4 3-5 3-6 Method option 2 2 3 3 3 3 Appearance Pale yellow solid Pale yellow solid Pale yellow solid Pale yellow solid Pale yellow solid Pale yellow solid Chiral purity >99% >99% >99.9% >99.9% >99.9% >99.9% Heavy metals < 20 ppm < 20 ppm < 5 ppm < 5 ppm < 5 ppm No analysis performed Residual solvent content (according to GC data) iPAC 4148 PPm DXM 8 PPm IPS 6 PPm Tol BUT. iPAc 7440 ppm DXM no. IPS 52 PPm Tol BUT. iPAc BUT. DXM NO. IPA 65 ppm Tol 2020 ppm iPAc BUT. DXM NO. IPS NO. Tol 3022 PPm iPAc BUT. DXM NO. IPA 109 ppm Tol 2020 ppm iPAc BUT. DXM NO. IPS NO. Tol 1337 PPm Water content (Karl-Fischer method) <0.1 wt. % 0.2 wt. % 0.2 wt. % 0.2 wt. % 0.3 wt. % 0.2 wt. % Particle size distribution (laser diffraction method) D(50) 140 µm D(50) 118 µm D(50) 91 µm D(50) 169 µm D(50) 122 µm D(50) 48 µm

N.O.=Undefined; iPAc=isopropyl acetate; DCM=dichloromethane; IPA=isopropyl alcohol; Tol=toluene.

- 55 046788

Table 4. HPLC Impurity Profile Comparison for Lots Prepared Using Method Options 2 and 3

Batch number 3-1 3-2 3-3 3-4 3-5 3-6 Method option 2 2 3 3 3 3 Analysis (wt.%, based on anhydrous substance) 99.3 97.9 99.9 96.7 97.8 98.4 Purity (%) 98.8 98.3 99.46 99.40 99.28 99.51 Identified impurities (%) RRT 0.38 (NT-316 without group Vos) (1-7) BUT. BUT. oh and 0.09 0.13 0.05 RRT 1.08 (imin) (1-1) 0.62 0.95 0.10 0.13 0.04 0.09 RRT 1.21 (aldehyde) (1-6) NI* NI* 0.09 0.14 0.20 0.14 RRT 1.28 (formyl) (1-2) 0.03 But. 0.03 0.03 0.03 0.05 RRT 1.37 (acetamide) (1-5) 0.07 0.42 0.02 0.04 0.06 BUT. RRT 1.93 (benzimidazole) (1-3) 0.01 BUT. 0.06 0.05 0.04 0.04 Total impurity content (%) 1.20 1.67 0.54 0.60 0.72 0.49

N.O.=Undefined.

N.I.=Not researched.

^The presence of any aldehyde impurities in these two batches was not determined.

Improved residual solvent content in method 3

When X4P-001 (formerly AMD110170) was first developed for clinical studies, the drug substance was directly isolated in free base form using ethyl acetate as a crystallization solvent (method 1). However, difficulties arose in removing ethyl acetate from the finished API, and obtaining test batches required labor-intensive processes involving grinding and hot nitrogen treatment to reduce its content below the limit set by the International Conference on Harmonization of Technical Requirements for Registration of Drugs for Human Use ( ICH), which is no more than 5000 ppm. Initial attempts to produce AMD-110170 for several batches of the API also used isopropyl acetate as the isolation solvent according to Method 1.

Later in the preparation process, the para-hydroxybenzoic acid (PHBA) salt of AMD11070 was isolated as an intermediate, followed by conversion to the free base (method 2). The inventors of the present invention used this method to produce drug substance for recent clinical trials. According to method 2, the free base was isolated after isolation of the salt using isopropyl acetate instead of ethyl acetate. This API exhibited similar high residual solvent content as the API produced by Method 1. In addition, the present inventors discovered that isopropyl acetate was involved in the formation of acetamide impurity during the production process. Thus, a change in solvent for isolating the finished API was justified.

A recent process improvement proposed by the present inventors provided a technique for direct isolation of the free base (the PHBA salt was no longer an intermediate in the preparation process) using toluene instead of isopropyl acetate as the final crystallization solvent (method 3). The present inventors found that toluene did not react with X4P-001 and was therefore a more suitable isolation solvent. As noted above, eliminating the use of isopropyl acetate also resulted in a reduction in impurity levels. Data on the residual content of solvents in batches of X4P-001 and the residual content of solvents for crystallization are summarized in table. 5.

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Table 5. Residual content of solvents for the isolation of X4P-001 in the process of improving the method

Solvent for isolation Determinable content (PPt)

Method No. 1

Ethyl acetate or isopropyl acetate 7219

Ethyl acetate or isopropylacetate 9108 Ethyl acetate or isopropylacetate 1971 Ethyl acetate or isopropylacetate 6820 Ethyl acetate or isopropylacetate 9298

Method No. 2

Isopropyl acetate 2500 Isopropyl acetate* * 4148** Isopropyl acetate 7440 Isopropyl acetate and toluene* N O. (iPAc)* 2280 (Toluene)*

Method No. 3

Toluene 4487 Toluene 2020 Toluene 3022 Toluene 2020 Toluene 1337

*A batch of API was processed from a batch that was initially crystallized using isopropyl acetate using toluene as a final crystallization step and was therefore analyzed for both isopropyl acetate (iPAc) and toluene.

**Game 3-1.

Initially, batches of X4P-001 were found to have average residual primary crystallization solvent contents ranging from 1971 to 9298 ppm. The present inventors used an acceptable daily exposure (ADE) approach to develop a specification for the residual toluene content of X4P-001 free base.

Based on a dose of X4P-001 of 600 mg per day (50% safety margin compared to the typical clinical dose of 400 mg per day), the estimated toluene content of X4P-001 API should not exceed 4500 ppm. Therefore, in some embodiments of the present invention, the specification for the residual toluene content of X4P-001 allows no more than 4500 ppm. Accordingly, some embodiments of the present invention provide a composition of X4P-001 containing no more than 4500 ppm toluene or 1350 ppm toluene.

conclusions

Overall, the above improvements presented in Process Version 3 provided a more reliable and reproducible final process step compared to Process Version 2. Most importantly, the purity of the isolated X4P-001 API was significantly improved compared to the previous Process Version. It is important to note that the present inventors have discovered that the choice of solvents during the preparation and isolation of X4P-001 is a key factor in improving the impurity profile described above. Specifically, the present inventors replaced the dichloromethane and isopropyl acetate used in the preparation and isolation of X4P-001 in Method Option 2 with 1-butanol and toluene/methanol, respectively, in Method Option 3. The present inventors found that In batches produced using Method Variant 2, some impurities were present in the finished product in increased quantities due to the reaction of the API, X4P-001, with dichloromethane and isopropyl acetate. Another key finding was that resin formation and API oiling during final product crystallization in Method Option 2 was associated with inconsistent (and often poor) quality of the AMD-2890 starting material. As described above, the present inventors introduced an improved AMD-2890 isolation technique that consistently provided high quality AMD-2890 and avoided gum formation and API oiling.

Example 10: Mutagenicity Assessment

The authors of the present invention have assessed the risk of formation of mutagenic substances associated with the synthetic method for producing X4P-001, all intermediate compounds related to this method, possible and detected impurities. The assessment included consideration of all raw materials, including key starting materials, as well as possible and detected impurities and degradation products generated during the production process. All assessments were carried out in accordance with the requirements of ICH Guide M7(R1).

Scheme 10 and Scheme 11 below present synthesis method 2 and synthesis method 3, respectively, for X4P-001. Scheme 12 shows the synthesis and resulting intermediates for NT319 and AMD-2890.

In table 6 below shows the detected potentially mutagenic impurities for the method of producing X4P-001.

Scheme 10: Synthesis method 2 for X4P-001

OO Stage 1a NH ₂ "2HCI * NT-272 OO Stage 1b ------ HCI"HN- ,B( ^v N Vos NT-316 'CO ' NYN ( Stage 3 1 --------► N ^x NH <M> Non-dedicated - intermediate - connection X4 P-001 Vos./h/xA ~ -] G η VOS Η [1 J 1 Ν^>^ Η Ί 1 ΝΤ-319 _κ -, _n II Λ J /\ _K1 ^Vos Σ Stage 1b _Vos nh ₂ ^reconnection 1 _ J Imine Intermediate Step 2 || xC | jc “ ΟΟΌ Cl ^N - I Г Ф—' N. ,,Boc /A/ (Ύ γ Vos Vos AMD-2890 L ^Vos _ tri-Boc-AMO-11070 with Stage 4 Stage 5 _______ p NH ₂ ___________in the form of free O N ^x NH । base b f X4R-001 ^C00N in the form of PHBA salt

- 58 046788

Scheme 11: Synthesis method 3 for X4P-001 n-butanol ^BoC'Y

Vos >o

N^Y

NH ₂ "2HCI

Stage 1a

NT-319

Ν'

N.

NT-272

N

NH ₂

Stage 1b

^N'B0C

Re-compound 1 Imine intermediate

Stage 1 b N τ'

----► HCkHN.

Stage 2

Stage For

NT-316

N' ^B0C

Vos

N,N.

• N Cl

Ν γ

N N.

N

Vos

AMD-2890 I-9

NH ₂

Stage 33

N

Vos

NY.N.

^N'B0C

Vos tri-Boc-AMD-l 1070

NH ₂

X4P-001 in free base form

X4P-001 in free base form

Scheme 12: Synthesis of NT-319 and AMD-2890.

OEt ^{Stage 10a}

OEt

ABA

Boc.N·H

OEt

OEt

Stage 10b

Vox^

Vos

OEt

OEt

4,4-diethoxybutane-1-amine

Connection 2

Connection 3

Stage 10c ^B ° ^C 'N ;O

Vos

NT-319

RM-396 I-8

C.I.

Stage 6

N

C.I.

N

Vos

AMD-2890 1-9

Table 6. Identified potentially mutagenic impurities for the X4P-001 synthesis method

The expected duration of treatment for patients participating in clinical trials is less than 10 years. The permissible intake of the drug for exposure > 1-10 years is 10 pg/day of mutagenic impurity according to Table. 2 in ICH Guide M7(R1).

Based on the X4P-001 dose of 400 mg/day, a limit of up to 25 ppm of potentially mutagenic impurities is acceptable (10 pg/day/400 mg = 25 ppm (p/g)).

Possible impurities I-10 and I-11 were not detected in X4P-001. Moreover, the theoretical calculation

- 59046788 purification factor for synthesis method 3, based on published sources such as Teasdale et al. (Org. Process Res. Dev. 2013, 17, 221), showed that this method provides effective removal of both possible impurities I-10 and I-11, while the theoretical purification coefficient is approximately 9x10 ⁸ (I-10) and approximately 9x10 ¹⁰ (I-11).

Based on the calculated purification factors, the theoretical residual amounts of I-10 and I-11 in X4P-001 were calculated to be approximately <0.0001 ppm I-10 and approximately <0.000001 ppm I11, indicating effective removal of both potentially mutagenic impurities using this method and without the need to control the purity of X4P-001.

Potential impurities RM-396 and AMD-2890 (I-9) were not detected (limit of quantitation, or LOQ, 2.5 ppm) in X4P-001, indicating that both potentially mutagenic impurities were effectively eliminated by this method.

Example 11 Solid formulations containing 25, 100 and 200 mg

During formulation, excipients were selected based on screening studies including short-term compatibility of various excipients. Microcrystalline cellulose was selected as the diluent/filler for the lower dosage capsules containing 25 mg. To increase flowability for higher dosage formulations containing 100 mg, a dibasic calcium phosphate dihydrate diluent/excipient was added to the formulation. The ratio of microcrystalline cellulose to dibasic calcium phosphate dihydrate was selected to approximate the bulk density of the drug substance to reduce the likelihood of segregation during the mixing process. Sodium stearyl fumarate was chosen as a lubricant. To facilitate filling of capsules using an automatic capsule filler, a glidant, colloidal silica, was added as a flow additive. Croscarmellose sodium was chosen as the disintegrant to ensure automatic capsule filling. To reduce the sticking of the drug substance when encapsulated, sodium lauryl sulfate was added to the composition as a wetting agent.

Table 7. Composition. in the form of standard capsules containing 25 mg X4P-001

Component Compliance with standard Purpose Amount (mg/capsule) % w/w Composition X4P-001 Own standard Active Ingredient 25.0 14.7 Microcrystalline cellulose NF Diluent 132.7 78.1 Croscarmellose sodium NF Baking powder 10.2 6.0 Sodium stearyl fumarate NF Lubricant 17 1.0 Colloidal silicon dioxide USP Flow additive 0.4 0.2 Total 170.0 100.0 Hard gelatin capsules, size 1 USP Package - -

- 60 046788

Table 8. Composition in the form of capsules containing 100 mg X4P-001

Component Compliance with standard Purpose 100 mg Amount (mg/capsule) wt./mass. Composition X4P-001 Own standard Active substance 100.0 37.6% Dibasic calcium phosphate dihydrate USP/NF Diluent 84.3 31.7% Microcrystalline cellulose NF/EP Diluent 60.9 22.9% Croscarmellose sodium NF/EP Baking powder 16.0 6.0% Sodium stearyl fumarate NF Lubricant 2.7 1.0% Sodium lauryl sulfate NF/EP Wetting agent 1.3 0.5% Colloidal silicon dioxide NF/EP Flow additive 0.8 0.3% Total 266.0 100 % Hard gelatin capsules, size 1 white/white. Qualitative composition: gelatin and titanium dioxide. USP Encapsulation - -

Table 9. Composition in the form of capsules containing 200 mg X4P-001

200 mg Ingredients Percentage per capsule (%) Theoretical amount per capsule (mg) Composition X4P-001 61.5 200.0 Microcrystalline cellulose, NF/EP (Avicel PH 101) or equivalent 12.9 41.93 Dibasic calcium phosphate dihydrate, USP/NF 17.8 57.85 Croscarmellose sodium, NF/EP (AcDi-Sol) 6.0 19.50 Sodium lauryl sulfate, NF/Ph. Eur. 0.5 1.625 Colloidal silicon dioxide, NF/Ph. Eur. (Cab-O-Sil M-5P) o, s 0.9750 Sodium stearyl fumarate, NF (Pruv) 1.0 3,250 Total contents of the capsule 100 325.0

Compatibility of excipients was demonstrated through ongoing long-term drug stability studies in which the drug met stability requirements under specific refrigerated storage conditions in 3 lots of 100 mg capsules. Stability study results are available for up to 24 months at storage conditions of 5°C±3°C and 25°C/60% RH for lot 15K227 capsules containing 100 mg X4P-001, packaged in 30-piece amber glass bottles. hermetically sealed in aluminum foil bags. Under the recommended storage conditions of 5°C ± 3°C, the results indicated no significant deviations in any parameters determined in the stability analysis for a period of up to 24 months (potency analysis, impurity content, solubility, moisture content and microbiological tests).

Results of a stability study over a period of up to 9 months for capsules containing 100 mg, stored at 5°C±3°C and packaged in amber glass vials sealed in aluminum foil pouches or high-density polyethylene vials ( HDPE) Oxy-Guard showed an increase in imine impurity over time, but the content remained within specification limits. No deviations from baseline were found for any other stability analysis parameters. Stability studies over a period of up to 3 months for capsules containing 100 mg stored at 5°C ± 3°C and 25°C/60% RH and packaged in Oxy-Guard HDPE bottles indicated an increase in imine impurity with over time, but the content remained within the limits allowed in the specification. No deviations from baseline were found for any other stability analysis parameters. These capsules were obtained using a medicinal substance

- 61 046788 tion X4R-001, in the production of which a processing technique using toluene was used.

The continued use of refrigerated storage in a cold chain system is an expected requirement for the drug product X4P-001. The primary packaging container for X4P-001 to be used in the Phase 3 clinical trial is a 60 cc ^HDPE Oxy-Guard vial with a 33 mm diameter cap with an induction sealed gasket. Rayon wadding is placed over the capsules in each bottle and one packet of desiccant (0.5 g Sorb-It or equivalent) is placed in the top of each bottle between the rayon wadding and the cap (30 pieces). A detailed description of the production method is presented in Fig. 1.

Claims (33)

1. Pharmaceutical composition X4P-001 containing a compound of formula I or a pharmaceutically acceptable salt thereof and the following compound: or a pharmaceutically acceptable salt thereof, in an amount of from 0.001 to 0.5% wt./wt. by weight of composition X4R-001. 2. Composition X4P-001 according to claim 1, where the amount of I-6 or a pharmaceutically acceptable salt thereof is from 0.01 to 0.4% w/w. by weight of composition X4R-001. 3. Composition X4P-001 according to claim 1, where the amount of I-6 or a pharmaceutically acceptable salt thereof is from 0.01 to 0.3% w/w. by weight of composition X4R-001. 4. Composition X4P-001 according to claim 1, where the amount of I-6 is from 0.01 to 0.2% wt./wt. by weight of composition X4R-001. 5. Composition X4P-001 according to any one of claims 1-4, containing at least the following compound: or a pharmaceutically acceptable salt thereof. 6. Composition X4P-001 according to any one of claims 1 to 5, containing at least the following compound: or a pharmaceutically acceptable salt thereof. 7. Composition X4P-001 according to any one of claims 1 to 6, containing at least the following compound: - 62 046788 1-5 or a pharmaceutically acceptable salt thereof. 8. Composition X4P-001 according to claim 6, containing at least the following compound: or a pharmaceutically acceptable salt thereof. 9. The X4P-001 composition according to claim 8, wherein said X4P-001 composition does not contain I-2 or I-5 or a pharmaceutically acceptable salt thereof. 10. Composition X4P-001 according to claim 5, where the amount of I-2 or a pharmaceutically acceptable salt thereof is from 0.01 to 0.3% w/w. by weight of composition X4R-001. 11. Composition X4P-001 according to claim 6 or 8, where the amount of I-3 or a pharmaceutically acceptable salt thereof is from 0.01 to 0.4% w/w. by weight of composition X4R-001. 12. Composition X4P-001 according to claim 7, where the amount of I-5 or a pharmaceutically acceptable salt thereof is from 0.01 to 0.4% w/w. by weight of composition X4R-001. 13. Composition X4P-001 according to claim 12, where the amount of I-7 or a pharmaceutically acceptable salt thereof is from 0.01 to 0.25% w/w. by weight of composition X4R-001. 14. Composition X4P-001 according to any one of claims 1 to 13, where the total mass of I-6 and any additional impurities present is no more than 0.8% wt./wt. by weight of composition X4R-001. 15. Pharmaceutical composition X4P-001 containing a compound of formula I I or a pharmaceutically acceptable salt thereof and at least one of the following compounds: 1-2 or a pharmaceutically acceptable salt thereof in an amount of from 0.01 to 0.3% wt./wt. by weight of the composition X4R-001; or a pharmaceutically acceptable salt thereof in an amount of from 0.01 to 0.4% w/w. from the mass of the components - 63 046788 position Х4Р-001; or a pharmaceutically acceptable salt thereof in an amount of from 0.01 to 0.4% w/w. by weight of the composition X4R-001; or a pharmaceutically acceptable salt thereof in an amount of from 0.01 to 0.4% w/w. by weight of the composition X4P-001 or or a pharmaceutically acceptable salt thereof in an amount of from 0.01 to 0.25% w/w. by weight of composition X4R-001. 16. Composition X4P-001 according to claim 15, not containing the following compound: or a pharmaceutically acceptable salt thereof. 17. Composition X4P-001 according to claim 15 or 16, containing each of I-2, I-3, I-5, I-6 and I-7 or a pharmaceutically acceptable salt thereof. 18. Composition X4P-001 according to any of paragraphs 15-16, additionally containing I-1 or a pharmaceutically acceptable salt thereof in an amount of from 0.01 to 0.7% w/w. by weight of composition X4R-001. 19. Composition X4P-001 according to any one of claims 15 to 18, where the amount of I-2 or a pharmaceutically acceptable salt thereof is from 0.01 to 0.2% w/w. by weight of composition X4R-001. 20. Composition X4P-001 according to any one of claims 15 to 19, where the amount of I-3 or a pharmaceutically acceptable salt thereof is from 0.01 to 0.2% w/w. by weight of composition X4R-001. 21. Composition X4P-001 according to any one of claims 15 to 20, where the amount of I-5 or a pharmaceutically acceptable salt thereof is from 0.01 to 0.2% w/w. by weight of composition X4R-001. 22. Composition X4P-001 according to any one of claims 15 to 21, where the amount of I-6 or a pharmaceutically acceptable salt thereof is from 0.01 to 0.2% w/w. by weight of composition X4R-001. 23. Composition X4P-001 according to any one of claims 15 to 22, where the amount of I-7 or a pharmaceutically acceptable salt thereof is from 0.01 to 0.2% w/w. compositions Х4Р-001. - 64 046788 24. Composition X4P-001 according to any one of claims 15 to 23, where the amount of I-1 or a pharmaceutically acceptable salt thereof is from 0.02 to 0.5% w/w. by weight of composition X4R-001. 25. Composition X4P-001 according to any one of claims 15-24, containing at least two compounds selected from: I-2 or a pharmaceutically acceptable salt thereof, I-3 or a pharmaceutically acceptable salt thereof, I-5 or a pharmaceutically acceptable salt thereof, I-6 or a pharmaceutically acceptable salt thereof or I-7 or a pharmaceutically acceptable salt thereof. 26. Composition X4P-001 according to any one of claims 15-25, containing at least three compounds selected from: I-2 or a pharmaceutically acceptable salt thereof, I-3 or a pharmaceutically acceptable salt thereof, I-5 or a pharmaceutically acceptable salt thereof, I-6 or a pharmaceutically acceptable salt thereof or I-7 or a pharmaceutically acceptable salt thereof. 27. Composition X4P-001 according to any one of claims 15-24, containing at least four compounds selected from: I-2 or a pharmaceutically acceptable salt thereof, I-3 or a pharmaceutically acceptable salt thereof, I-5 or a pharmaceutically acceptable salt thereof, I-6 or a pharmaceutically acceptable salt thereof or I-7 or a pharmaceutically acceptable salt thereof. 28. Composition X4P-001 according to any one of claims 1 to 14, where said composition X4P-001 does not contain the following compound: 1-4 or a pharmaceutically acceptable salt thereof. 29. Composition X4P-001 according to any one of claims 1 to 28, wherein the compound of formula I is present in said composition X4P-001 as a free base. 30. Composition X4P-001 according to any one of claims 1-29, in which the total impurity content is no more than 3.0%. 31. Composition X4P-001 according to any one of claims 1 to 30, in which the enantiomeric excess (% ee) of the R enantiomer is at least 97.0%. 32. A pharmaceutical composition comprising the composition X4P-001 according to any one of claims 1 to 31 and a pharmaceutically acceptable adjuvant, carrier or transfer medium. 33. Compound of the following formula: 1-6 or a pharmaceutically acceptable salt thereof.