Patent Description:
Antibiotics are the most effective drugs for treating bacteria-infectious diseases. They are largely used in the clinic because of their good antibacterial effect with limited side effects. Among them, the beta-lactam class of antibiotics (for example, penicillins, cephalosporins, monobactams and carbapenems) are preferred because their effect is bactericidal and their target is absent in eukaryotic cells with consequent low toxicity.

To counter the efficacy of the various beta-lactams, bacteria have evolved to produce variants of beta-lactam deactivating enzymes called beta-lactamases, and in the ability to share this tool both vertically and horizontally inter- and intra-species. These beta-lactamases are categorized as "serine" or "metallo" based, respectively, based on the presence of a key serine or zinc in the enzyme active site. The rapid induction, selection and spread of this mechanism of bacterial resistance can severely limit the whole class of beta-lactam treatment options in the hospital and in the community. There is a need for effective and safe therapeutic agents that can treat such resistant infections.

<CIT>) describes certain compounds and compositions that modulate the activity of beta-lactamases and their use in the treatment of bacterial infections.

One aspect of the invention is a crystalline form of (R)-<NUM>-(<NUM>-(trans-<NUM>-(<NUM>-aminoethylamino)cyclohexyl)acetamido)-<NUM>-hydroxy-<NUM>,<NUM>-dihydro-<NUM>-benzo[e][<NUM>,<NUM>]oxaborinine-<NUM>-carboxylic acid dihydrochloride:
<CHM>.

The crystalline form, (R)-<NUM>-(<NUM>-(trans-<NUM>-(<NUM>-aminoethylamino)cyclohexyl)acetamido)-<NUM>-hydroxy-<NUM>,<NUM>-dihydro-<NUM>-benzo[e][<NUM>,<NUM>]oxaborinine-<NUM>-carboxylic acid dihydrochloride, is anhydrous.

In some embodiments, the crystalline form has an X-Ray powder diffraction (XRPD) pattern substantially the same as shown in <FIG>.

In some embodiments, the crystalline form has an X-ray powder diffraction (XRPD) pattern comprising characteristic peaks at about <NUM>°±<NUM>. <NUM>° 2θ, about <NUM>°±<NUM>° 2θ, about <NUM>°±<NUM>° 2θ, about <NUM>°±<NUM>° 2θ, about <NUM>°±<NUM>° 2θ, and about <NUM>°±<NUM>° 2θ.

In some embodiments of the crystalline form, the X-ray powder diffraction (XRPD) pattern further comprises characteristic peaks at about <NUM>°±<NUM>° 2θ, about <NUM>°±<NUM>° 2θ, about <NUM>°±<NUM>° 2θ, about <NUM>°±<NUM>° 2θ, about <NUM>°±<NUM>° 2θ, and about <NUM>°±<NUM>° 2θ.

Another aspect of the invention is a pharmaceutical composition comprising:.

In some embodiments of a pharmaceutical composition, the pharmaceutical composition is formulated as a homogeneous liquid suitable for injection.

In some embodiments of a pharmaceutical composition, the pharmaceutical composition further comprises an aqueous carrier.

In some embodiments of a pharmaceutical composition, the pharmaceutical composition has a pH from about <NUM> to about <NUM>.

In some embodiments of a pharmaceutical composition, the pharmaceutical composition is formulated as a powder for reconstitution.

In some embodiments of a pharmaceutical composition, the pharmaceutical composition is suitable for injection once reconstituted with an aqueous carrier.

In some embodiments of a pharmaceutical composition, the pharmaceutical composition further comprises a pharmaceutically acceptable excipient.

In some embodiments of a pharmaceutical composition, the pharmaceutically acceptable excipient is an amino acid or a monosaccharide derivative.

In some embodiments of a pharmaceutical composition, the pharmaceutically acceptable excipient is L-arginine.

In some embodiments of a pharmaceutical composition, the pharmaceutically acceptable excipient is meglumine.

The novel features of the disclosure are set forth with particularity in the appended claims. A better understanding of the features and advantages of the present disclosure will be obtained by reference to the following detailed description that sets forth illustrative embodiments, in which the principles of the disclosure are utilized, and the accompanying drawings of which:.

Any references herein to methods of treatment refer to the compounds, pharmaceutical compositions, and medicaments of the present invention for use in a such methods.

Described herein are crystalline forms of (R)-<NUM>-(<NUM>-(trans-<NUM>-(<NUM>-aminoethylamino)cyclohexyl)acetamido)-<NUM>-hydroxy-<NUM>,<NUM>-dihydro-<NUM>-benzo[e][<NUM>,<NUM>]oxaborinine-<NUM>-carboxylic acid dihydrochloride.

Also described herein are pharmaceutical compositions comprising crystalline forms of (R)-<NUM>-(<NUM>-(trans-<NUM>-(<NUM>-aminoethylamino)cyclohexyl)acetamido)-<NUM>-hydroxy-<NUM>,<NUM>-dihydro-<NUM>-benzo[e][<NUM>,<NUM>]oxaborinine-<NUM>-carboxylic acid dihydrochloride.

As used herein, (R)-<NUM>-(<NUM>-(trans-<NUM>-(<NUM>-aminoethylamino)cyclohexyl)acetamido)-<NUM>-hydroxy-<NUM>,<NUM>-dihydro-<NUM>-benzo[e][<NUM>,<NUM>]oxaborinine-<NUM>-carboxylic acid is as shown in the structure below:
<CHM>
(R)-<NUM>-(<NUM>-(trans-<NUM>-(<NUM>-aminoethylamino)cyclohexyl)acetamido)-<NUM>-hydroxy-<NUM>,<NUM>-dihydro-<NUM>-benzo[e][<NUM>,<NUM>]oxaborinine-<NUM>-carboxylic acid is also referred to as Compound <NUM>. Compound <NUM> may exist in an equilibrium between the "closed" cyclic form (as shown above) and the "open" acyclic form:
<CHM>
Compound <NUM> may associate into intramolecular dimers, trimers, and any combinations thereof.

The dihydrochloride salt of Compound <NUM> can be made into a pure and stable crystalline form (Form <NUM>) under controlled conditions. In some embodiments, Form <NUM> is a monohydrate form of the dihydrochloride salt of Compound <NUM>. In some embodiments, the X-ray powder diffraction (XRPD) pattern of Form <NUM> is substantially the same as shown in <FIG>, with corresponding tabulated peak data shown in Table <NUM>. In some embodiments, the X-ray powder diffraction (XRPD) pattern of Form <NUM> is substantially the same as shown in <FIG>. The X-ray powder diffraction (XRPD) pattern shown in <FIG> is substantially the same as the X-ray powder diffraction (XRPD) pattern shown in <FIG>.

The dihydrochloride salt of Compound <NUM> can be made into a pure and stable crystalline form (Form <NUM>) under controlled conditions. Form <NUM> is an anhydrous form of the dihydrochloride salt of Compound <NUM>. The X-ray powder diffraction (XRPD) pattern of Form <NUM> is substantially the same as shown in <FIG>, with corresponding tabulated peak data shown in Table <NUM>.

Form <NUM> exhibits an X-ray powder diffraction pattern that includes at least three characteristic peaks selected from peaks at about <NUM>°±<NUM>° <NUM>, about <NUM>°±<NUM>° <NUM>, about <NUM>°±<NUM>° <NUM>, about <NUM>°±<NUM>° <NUM>, about <NUM>°±<NUM>° <NUM>, at about <NUM>°±<NUM>° <NUM>, about <NUM>°±<NUM>° <NUM>, about <NUM>°±<NUM>° <NUM>, about <NUM>°±<NUM>° 2θ,about <NUM>°±<NUM>° 2θ, about <NUM>°±<NUM>° 2θ, and about <NUM>°±<NUM>° 2θ. In some embodiments, Form <NUM> exhibits an X-ray powder diffraction (XRPD) pattern that comprises characteristic peaks at about <NUM>°±<NUM>° <NUM>, about <NUM>°±<NUM>° <NUM>, about <NUM>°±<NUM>° <NUM>, about <NUM>°±<NUM>° <NUM>, about <NUM>°±<NUM>° 2θ,and about <NUM>°±<NUM>° 2θ. In some embodiments, Form <NUM> exhibits an X-ray powder diffraction (XRPD) pattern that further comprises characteristic peaks at about <NUM>°±<NUM>° <NUM>, about <NUM>°±<NUM>° <NUM>, about <NUM>°±<NUM>° <NUM>, about <NUM>°±<NUM>° 2θ, about <NUM>°±<NUM>° <NUM>, and about <NUM>°±<NUM>° 2θ. In some embodiments, Form <NUM> exhibits an X-ray powder diffraction (XRPD) pattern that comprises characteristic peaks at about <NUM>°±<NUM>° <NUM>, about <NUM>°±<NUM>° <NUM>, about <NUM>°±<NUM>° <NUM>, about <NUM>°±<NUM>° <NUM>, about <NUM>°±<NUM>° <NUM>, about <NUM>°±<NUM>° <NUM>, about <NUM>°±<NUM>° <NUM>, about <NUM>. <NUM>°±<NUM>° 2θ, about <NUM>°±<NUM>° 2θ, about <NUM>°±<NUM>° 2θ, about <NUM>°±<NUM>° 2θ, and about <NUM>°±<NUM>° <NUM>. In some embodiments, Form <NUM> exhibits an X-ray powder diffraction (XRPD) pattern that comprises characteristic peaks at about <NUM>°±<NUM>° <NUM>, about <NUM>°±<NUM>° <NUM>, about <NUM>°±<NUM>° <NUM>, about <NUM>°±<NUM>° 2θ, about <NUM>°±<NUM>° 2θ, about <NUM>°±<NUM>° 2θ, about <NUM>°±<NUM>° 2θ, about <NUM>°±<NUM>° 2θ, about <NUM>°±<NUM>° 2θ, about <NUM>°±<NUM>° 2θ, about <NUM>°±<NUM>° 2θ, about <NUM>°±<NUM>° <NUM>, about <NUM>°±<NUM>° <NUM>, about <NUM>°±<NUM>° <NUM>, about <NUM>°±<NUM>° <NUM>, about <NUM>°±<NUM>° <NUM>, about <NUM>°±<NUM>° <NUM>, about <NUM>°±<NUM>° 2θ, about <NUM>°±<NUM>° 2θ, and about <NUM>°±<NUM>° 2θ.

The dihydrochloride salt of Compound <NUM> can be made into a new form (Form <NUM>) under controlled conditions. The X-ray powder diffraction (XRPD) pattern of Form <NUM> is substantially the same as shown in <FIG>, with corresponding tabulated peak data shown in Table <NUM>.

In some embodiments, the dihydrochloride salt of Compound <NUM> can be made into a crystalline form (Form <NUM>) under controlled conditions. The X-ray powder diffraction (XRPD) pattern of Form <NUM> is substantially the same as shown in <FIG>, with corresponding tabulated peak data shown in Table <NUM>.

A first aspect of the invention is a crystalline form of (R)-<NUM>-(<NUM>-(trans-<NUM>-(<NUM>-aminoethylamino)cyclohexyl)acetamido)-<NUM>-hydroxy-<NUM>,<NUM>-dihydro-<NUM>-benzo[e][<NUM>,<NUM>]oxaborinine-<NUM>-carboxylic acid dihydrochloride:
<CHM>.

The crystalline form of (R)-<NUM>-(<NUM>-(trans-<NUM>-(<NUM>-aminoethylamino)cyclohexyl)acetamido)-<NUM>-hydroxy-<NUM>,<NUM>-dihydro-<NUM>-benzo[e][<NUM>,<NUM>]oxaborinine-<NUM>-carboxylic acid dihydrochloride is anhydrous.

In some embodiments, the crystalline form has at least one of the following properties:.

In some embodiments, the X-ray powder diffraction (XRPD) pattern further comprises characteristic peaks at about <NUM>°±<NUM>° <NUM>, about <NUM>°±<NUM>° <NUM>, about <NUM>°±<NUM>° <NUM>, about <NUM>°±<NUM>° <NUM>, about <NUM>°±<NUM>° <NUM>, and about <NUM>°±<NUM>° 2θ.

In some embodiments, the crystalline form is Form <NUM>.

Again, any references herein to methods of treatment refer to the compounds, pharmaceutical compositions, and medicaments of the present invention for use in a such methods.

A second aspect of the invention is a pharmaceutical composition comprising:.

In some embodiments, the crystalline form of (R)-<NUM>-(<NUM>-(trans-<NUM>-(<NUM>-aminoethylamino)cyclohexyl)acetamido)-<NUM>-hydroxy-<NUM>,<NUM>-dihydro-<NUM>-benzo[e][<NUM>,<NUM>]oxaborinine-<NUM>-carboxylic acid dihydrochloride is Form <NUM>.

In some embodiments, the pharmaceutical compositions described herein are provided in unit dosage form. As used herein, a "unit dosage form" is a composition containing an amount of compound <NUM> that is suitable for administration to an animal, preferably mammal, subject in a single dose, according to good medical practice. The preparation of a single or unit dosage form however, does not imply that the dosage form is administered once per day or once per course of therapy. Such dosage forms are contemplated to be administered once, twice, thrice or more per day and may be administered as infusion over a period of time (e.g., from about <NUM> minutes to about <NUM>-<NUM> hours), or administered as a continuous infusion, and may be given more than once during a course of therapy, though a single administration is not specifically excluded. In some embodiments, Compound <NUM> and cefepime are formulated in separate containers. In some embodiments, Compound <NUM> and cefepime are formulated in a single container.

In some embodiments, administration is via any of the accepted modes of administration for agents that serve similar utilities including, but not limited to, orally, subcutaneously, intravenously, intranasally, topically, transdermally, intraperitoneally, intramuscularly, intrapulmonarilly, vaginally, rectally, or intraocularly. In some embodiments, the administration is oral administration. In some embodiments, the administration is intravenous administration. In some embodiments, the administration is intramuscular administration.

Standard pharmaceutical formulation techniques are used, such as those disclosed in <NPL>).

In some embodiments, the pharmaceutical composition comprises a pharmaceutically acceptable excipient. The term "pharmaceutically acceptable excipient", as used herein, means one or more compatible solid or encapsulating substances, which are suitable for administration to a mammal. The term "compatible", as used herein, means that the components of the composition are capable of being commingled with the subject compound, and with each other, in a manner such that there is no interaction, which would substantially reduce the pharmaceutical efficacy of the composition under ordinary use situations. In some embodiments, the pharmaceutically acceptable excipient is of sufficiently high purity and sufficiently low toxicity to render them suitable for administration preferably to an animal, preferably mammal, being treated.

Some examples of substances, which can serve as pharmaceutically acceptable excipients include:.

In some embodiments of a pharmaceutical composition, the pharmaceutical composition is formulated as a homogeneous liquid suitable for injection. In some embodiments, the pharmaceutical composition is formulated for intravenous/intramuscular administration. In some embodiments of a pharmaceutical composition, the composition further comprises a pharmaceutically acceptable excipient. In some embodiments of a pharmaceutical composition, the pharmaceutically acceptable excipient is an amino acid, a monosaccharide, or a monosaccharide derivative. In some embodiments of a pharmaceutical composition, the pharmaceutically acceptable excipient is an amino acid or a monosaccharide derivative. In some embodiments of a pharmaceutical composition, the pharmaceutically acceptable excipient is an amino acid. In some embodiments of a pharmaceutical composition, the pharmaceutically acceptable excipient is arginine. In some embodiments of a pharmaceutical composition, the pharmaceutically acceptable excipient is L-arginine. In some embodiments of a pharmaceutical composition, the pharmaceutically acceptable excipient is a monosaccharide or a monosaccharide derivative. In some embodiments of a pharmaceutical composition, the pharmaceutically acceptable excipient is a monosaccharide derivative. In some embodiments of a pharmaceutical composition, the pharmaceutically acceptable excipient is meglumine.

In some embodiments, the pharmaceutical composition for intravenous/intramuscular administration is formulated as a homogeneous liquid suitable for injection. In some embodiments, the pharmaceutical composition further comprises an aqueous carrier. In some embodiments, the aqueous carrier is water for injection, <NUM>% sodium chloride injection, <NUM>% dextrose injection, <NUM>% dextrose injection, sodium lactate injection, <NUM>% dextrose and <NUM>% sodium chloride injection, lactated Ringers and <NUM>% dextrose injection, sodium chloride/sodium acetate/sodium gluconate/potassium chloride/magnesium chloride injection, sodium chloride/potassium acetate/magnesium acetate in <NUM>% dextrose injection, or any combinations thereof. In some embodiments, the aqueous carrier is water for injection. In some embodiments, the aqueous carrier is <NUM>% sodium chloride injection.

In some embodiments, the pH of the homogeneous liquid is from about <NUM> to about <NUM>. In some embodiments, the pH of the homogeneous liquid is about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, or about <NUM>. In some embodiments, the pH of the homogeneous liquid is from about <NUM> to about <NUM>.

In some embodiments, the pharmaceutical composition for intravenous/intramuscular administration is formulated as a powder for reconstitution. In some embodiments, the pharmaceutical composition is suitable for injection once reconstituted with an aqueous carrier.

In some embodiments, the aqueous carrier is water for injection, <NUM>% sodium chloride injection, <NUM>% dextrose injection, <NUM>% dextrose injection, sodium lactate injection, <NUM>% dextrose and <NUM>% sodium chloride injection, lactated Ringers and <NUM>% dextrose injection, sodium chloride/sodium acetate/sodium gluconate/potassium chloride/magnesium chloride injection, sodium chloride/potassium acetate/magnesium acetate in <NUM>% dextrose injection, or any combinations thereof.

The pharmaceutical compositions described herein are stable in various storage conditions including refrigerated, ambient, and accelerated conditions. In some embodiments, refrigerated condition is about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, or about <NUM>. Accelerated conditions include temperature and/or relative humidity (RH) that are at or above ambient levels (e.g. <NUM> ± <NUM>/<NUM>% RH ± <NUM>% RH). In some instances, an accelerated condition is at about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, about <NUM>, or about <NUM>. In other instances, an accelerated condition is above <NUM>% RH, about <NUM> % RH, about <NUM> % RH, about <NUM> % RH, or about <NUM> % RH. In further instances, an accelerated condition is about <NUM> or <NUM> at ambient humidity. In yet further instances, an accelerated condition is about <NUM> ± <NUM>/<NUM>% RH ± <NUM>% RH. In yet further instances, an accelerated condition is about <NUM> ± <NUM>/<NUM>% RH ± <NUM>% RH.

In some embodiments, the pharmaceutical composition is stable at about <NUM> ± <NUM> for at least <NUM> months. In some embodiments, the pharmaceutical composition is stable at about <NUM> ± <NUM>/<NUM>% RH ± <NUM>% RH for at least <NUM> months. In some embodiments, the pharmaceutical composition is stable at about <NUM> ± <NUM>/<NUM>% RH ± <NUM>% RH for at least <NUM> months. In some embodiments, the pharmaceutical composition is stable at about <NUM> ± <NUM>/<NUM>% RH ± <NUM>% RH for at least <NUM> months.

In some embodiments, the pharmaceutical composition is formulated as a homogeneous liquid suitable for injection.

In some embodiments, the pharmaceutical composition further comprises a pharmaceutically acceptable excipient.

In some embodiments, the pharmaceutically acceptable excipient is selected from amino acids.

In some embodiments, the amino acid is L-arginine.

In some embodiments, the pharmaceutical composition further comprises an aqueous carrier.

In some embodiments, the aqueous carrier is selected from water for injection, <NUM>% sodium chloride injection, <NUM>% dextrose injection, <NUM>% dextrose injection, sodium lactate injection, <NUM>% dextrose and <NUM>% sodium chloride injection, lactated Ringers and <NUM>% dextrose injection, sodium chloride/sodium acetate/sodium gluconate/potassium chloride/magnesium chloride injection, sodium chloride/potassium acetate/magnesium acetate in <NUM>% dextrose injection, and any combinations thereof.

In some embodiments, the pharmaceutical composition has a pH from about <NUM> to about <NUM>.

In some embodiments the pharmaceutical composition is formulated as a powder for reconstitution.

In some embodiments, the pharmaceutical composition is suitable for injection once reconstituted with an aqueous carrier.

In some embodiments, the pharmaceutical composition is stable at about <NUM> ± <NUM> for at least <NUM> months.

In some embodiments, the pharmaceutical composition is stable at about <NUM> ± <NUM>/<NUM>% RH ± <NUM>% RH for at least <NUM> months.

Also described herein is a method of administering to a subject in need thereof, a pharmaceutical composition comprising:.

The (R)-<NUM>-(<NUM>-(trans-<NUM>-(<NUM>-aminoethylamino)cyclohexyl)acetamido)-<NUM>-hydroxy-<NUM>,<NUM>-dihydro-<NUM>-benzo[e][<NUM>,<NUM>]oxaborinine-<NUM>-carboxylic acid dihydrochloride and the cefepime may be formulated in separate containers.

The (R)-<NUM>-(<NUM>-(trans-<NUM>-(<NUM>-aminoethylamino)cyclohexyl)acetamido)-<NUM>-hydroxy-<NUM>,<NUM>-dihydro-<NUM>-benzo[e][<NUM>,<NUM>]oxaborinine-<NUM>-carboxylic acid dihydrochloride may be formulated in a first container.

The (R)-<NUM>-(<NUM>-(trans-<NUM>-(<NUM>-aminoethylamino)cyclohexyl)acetamido)-<NUM>-hydroxy-<NUM>,<NUM>-dihydro-<NUM>-benzo[e][<NUM>,<NUM>]oxaborinine-<NUM>-carboxylic acid dihydrochloride may be in the form of a first powder for reconstitution.

The cefepime may be formulated in a second container.

The cefepime may be in the form of a second powder for reconstitution. The method may comprise:.

Each of the aqueous carriers may be independently selected from water for injection, <NUM>% sodium chloride for injection, <NUM>% dextrose for injection, <NUM>% dextrose for injection, sodium lactate for injection, <NUM>% dextrose and <NUM>% sodium chloride for injection, lactated Ringers and <NUM>% dextrose for injection, sodium chloride/sodium acetate/sodium gluconate/potassium chloride/magnesium chloride for injection, sodium chloride/potassium acetate/magnesium acetate in <NUM>% dextrose for injection, and any combinations thereof.

The (R)-<NUM>-(<NUM>-(trans-<NUM>-(<NUM>-aminoethylamino)cyclohexyl)acetamido)-<NUM>-hydroxy-<NUM>,<NUM>-dihydro-<NUM>-benzo[e][<NUM>,<NUM>]oxaborinine-<NUM>-carboxylic acid dihydrochloride, and the cefepime may be formulated in a single container.

The (R)-<NUM>-(<NUM>-(trans-<NUM>-(<NUM>-aminoethylamino)cyclohexyl)acetamido)-<NUM>-hydroxy-<NUM>,<NUM>-dihydro-<NUM>-benzo[e][<NUM>,<NUM>]oxaborinine-<NUM>-carboxylic acid dihydrochloride may be in the form of a powder for reconstitution.

The aqueous carrier may be selected from water for injection, <NUM>% sodium chloride injection, <NUM>% dextrose injection, <NUM>% dextrose injection, sodium lactate injection, <NUM>% dextrose and <NUM>% sodium chloride injection, lactated Ringers and <NUM>% dextrose injection, sodium chloride/sodium acetate/sodium gluconate/potassium chloride/magnesium chloride injection, sodium chloride/potassium acetate/magnesium acetate in <NUM>% dextrose injection, and any combinations thereof.

The pharmaceutical composition may further comprise a pharmaceutically acceptable excipient.

The pharmaceutically acceptable excipient may be selected from amino acids.

The pharmaceutically acceptable excipient may be L-arginine.

Also described herein is a method of treating a bacterial infection in a subject in need thereof, comprising administering to the subject a pharmaceutical composition as described herein.

The pharmaceutical composition may comprise about <NUM> of (R)-<NUM>-(<NUM>-(trans-<NUM>-(<NUM>-aminoethylamino)cyclohexyl)acetamido)-<NUM>-hydroxy-<NUM>,<NUM>-dihydro-<NUM>-benzo[e][<NUM>,<NUM>]oxaborinine-<NUM>-carboxylic acid.

The pharmaceutical composition may comprise about <NUM> of cefepime.

The pharmaceutical composition may be administered by intravenous (IV) infusion.

A period of infusion of the pharmaceutical composition may be about <NUM> hours.

In some embodiments, the pharmaceutical composition is administered via intravenous/intramuscular injection. In some embodiments, the pharmaceutical composition is infused into the patient over a period of time. In various embodiments, the infusion time (period of infusion) ranges from <NUM> minutes to continuous infusion, from <NUM> minutes to <NUM> hours, from <NUM> minutes to <NUM> hours, and from <NUM> hour to <NUM> hours. In one embodiment, the pharmaceutical composition is infused over a <NUM> minute period. In one embodiment, the pharmaceutical composition is infused over a <NUM> hour period. In one embodiment, the pharmaceutical composition is infused over a <NUM> hour period. In one embodiment, the pharmaceutical composition is infused over a <NUM> hour period. In one embodiment, the pharmaceutical composition is infused over a <NUM> hour period. In one embodiment, the pharmaceutical composition is infused over a <NUM> hour period. In some embodiments, the infusion is repeated at the desired dose interval, which includes, for example, <NUM> hours, <NUM> hours, <NUM> hours, or <NUM> hours.

In some embodiments, the pharmaceutical composition is administered for a period of one day. In some embodiments, the pharmaceutical composition is administered over a period of two days. In some embodiments, the pharmaceutical composition is administered over a period of three days. In some embodiments, the pharmaceutical composition is administered over a period of four days. In some embodiments, the pharmaceutical composition is administered over a period of five days. In some embodiments, the pharmaceutical composition is administered over a period of six days. In some embodiments, the pharmaceutical composition is administered over a period of seven days. In some embodiments, the pharmaceutical composition is administered over a period of eight days. In some embodiments, the pharmaceutical composition is administered over a period of nine days. In some embodiments, the pharmaceutical composition is administered over a period of ten days. In some embodiments, the pharmaceutical composition is administered over a period of <NUM> days. In some embodiments, the pharmaceutical composition is administered over a period of <NUM> days. In some embodiments, the pharmaceutical composition is administered over a period of <NUM> days. In some embodiments, the pharmaceutical composition is administered over a period of <NUM> days. In some embodiments, the pharmaceutical composition is administered over a period of <NUM> days. In some embodiments, the pharmaceutical composition is administered over a period of <NUM> days or more.

In some embodiments, the infusion is repeated at the desired dose interval, which includes, for example, <NUM> hours, <NUM> hours, <NUM> hours, or <NUM> hours for a period of one day. In some embodiments, the infusion is repeated at the desired dose interval, which includes, for example, <NUM> hours, <NUM> hours, <NUM> hours, or <NUM> hours over a period of two days. In some embodiments, the infusion is repeated at the desired dose interval, which includes, for example, <NUM> hours, <NUM> hours, <NUM> hours, or <NUM> hours over a period of three days. In some embodiments, the infusion is repeated at the desired dose interval, which includes, for example, <NUM> hours, <NUM> hours, <NUM> hours, or <NUM> hours over a period of four days. In some embodiments, the infusion is repeated at the desired dose interval, which includes, for example, <NUM> hours, <NUM> hours, <NUM> hours, or <NUM> hours over a period of five days. In some embodiments, the infusion is repeated at the desired dose interval, which includes, for example, <NUM> hours, <NUM> hours, <NUM> hours, or <NUM> hours over a period of six days. In some embodiments, the infusion is repeated at the desired dose interval, which includes, for example, <NUM> hours, <NUM> hours, <NUM> hours, or <NUM> hours over a period of seven days. In some embodiments, the infusion is repeated at the desired dose interval, which includes, for example, <NUM> hours, <NUM> hours, <NUM> hours, or <NUM> hours over a period of eight days. In some embodiments, the infusion is repeated at the desired dose interval, which includes, for example, <NUM> hours, <NUM> hours, <NUM> hours, or <NUM> hours over a period of nine days. In some embodiments, the infusion is repeated at the desired dose interval, which includes, for example, <NUM> hours, <NUM> hours, <NUM> hours, or <NUM> hours over a period of ten days. In some embodiments, the infusion is repeated at the desired dose interval, which includes, for example, <NUM> hours, <NUM> hours, <NUM> hours, or <NUM> hours over a period of <NUM> days. In some embodiments, the infusion is repeated at the desired dose interval, which includes, for example, <NUM> hours, <NUM> hours, <NUM> hours, or <NUM> hours over a period of <NUM> days. In some embodiments, the infusion is repeated at the desired dose interval, which includes, for example, <NUM> hours, <NUM> hours, <NUM> hours, or <NUM> hours over a period of <NUM> days. In some embodiments, the infusion is repeated at the desired dose interval, which includes, for example, <NUM> hours, <NUM> hours, <NUM> hours, or <NUM> hours over a period of <NUM> days. In some embodiments, the infusion is repeated at the desired dose interval, which includes, for example, <NUM> hours, <NUM> hours, <NUM> hours, or <NUM> hours over a period of <NUM> days. In some embodiments, the infusion is repeated at the desired dose interval, which includes, for example, <NUM> hours, <NUM> hours, <NUM> hours, or <NUM> hours over a period of <NUM> days or more. In certain embodiments, only a single infusion is administered on the last day for the treatment period.

In some embodiments, the infusion is repeated every <NUM> hours for a period of one day. In some embodiments, the infusion is repeated every <NUM> hours over a period of two days. In some embodiments, the infusion is repeated every <NUM> hours over a period of three days. In some embodiments, the infusion is repeated every <NUM> hours over a period of four days. In some embodiments, the infusion is repeated every <NUM> hours over a period of five days. In some embodiments, the infusion is repeated every <NUM> hours over a period of six days. In some embodiments, the infusion is repeated every <NUM> hours over a period of seven days. In some embodiments, the infusion is repeated every <NUM> hours over a period of eight days. In some embodiments, the infusion is repeated every <NUM> hours over a period of nine days. In some embodiments, the infusion is repeated every <NUM> hours over a period of ten days. In some embodiments, the infusion is repeated every <NUM> hours over a period of <NUM> days. In some embodiments, the infusion is repeated every <NUM> hours over a period of <NUM> days. In some embodiments, the infusion is repeated every <NUM> hours over a period of <NUM> days. In some embodiments, the infusion is repeated every <NUM> hours over a period of <NUM> days. In some embodiments, the infusion is repeated every <NUM> hours over a period of <NUM> days. In some embodiments, the infusion is repeated every <NUM> hours over a period of <NUM> days or more. In some embodiments, only one infusion is administered on the last day of the administration period. In some embodiments, the infusion is repeated every <NUM> hours over a period of <NUM> days with three doses on days <NUM>-<NUM> and one dose on day <NUM>.

In some embodiments, the infusion is repeated every <NUM> hours for a period of one day. In some embodiments, the infusion is repeated every <NUM> hours over a period of two days. In some embodiments, the infusion is repeated every <NUM> hours over a period of three days. In some embodiments, the infusion is repeated every <NUM> hours over a period of four days. In some embodiments, the infusion is repeated every <NUM> hours over a period of five days. In some embodiments, the infusion is repeated every <NUM> hours over a period of six days. In some embodiments, the infusion is repeated every <NUM> hours over a period of seven days. In some embodiments, the infusion is repeated every <NUM> hours over a period of eight days. In some embodiments, the infusion is repeated every <NUM> hours over a period of nine days. In some embodiments, the infusion is repeated every <NUM> hours over a period of ten days. In some embodiments, the infusion is repeated every <NUM> hours over a period of <NUM> days. In some embodiments, the infusion is repeated every <NUM> hours over a period of <NUM> days. In some embodiments, the infusion is repeated every <NUM> hours over a period of <NUM> days. In some embodiments, the infusion is repeated every <NUM> hours over a period of <NUM> days. In some embodiments, the infusion is repeated every <NUM> hours over a period of <NUM> days. In some embodiments, the infusion is repeated every <NUM> hours over a period of <NUM> days or more. In some embodiments, only one infusion is administered on the last day of the administration period.

In some embodiments, the infusion is repeated every <NUM> hours for a period of one day. In some embodiments, the infusion is repeated every <NUM> hours over a period of two days. In some embodiments, the infusion is repeated every <NUM> hours over a period of three days. In some embodiments, the infusion is repeated every <NUM> hours over a period of four days. In some embodiments, the infusion is repeated every <NUM> hours over a period of five days. In some embodiments, the infusion is repeated every <NUM> hours over a period of six days. In some embodiments, the infusion is repeated every <NUM> hours over a period of seven days. In some embodiments, the infusion is repeated every <NUM> hours over a period of eight days. In some embodiments, the infusion is repeated every <NUM> hours over a period of nine days. In some embodiments, the infusion is repeated every <NUM> hours over a period of ten days. In some embodiments, the infusion is repeated every <NUM> hours over a period of <NUM> days. In some embodiments, the infusion is repeated every <NUM> hours over a period of <NUM> days. In some embodiments, the infusion is repeated every <NUM> hours over a period of <NUM> days. In some embodiments, the infusion is repeated every <NUM> hours over a period of <NUM> days. In some embodiments, the infusion is repeated every <NUM> hours over a period of <NUM> days. In some embodiments, the infusion is repeated every <NUM> hours over a period of <NUM> days or more.

In some embodiments, the infusion is repeated every <NUM> hours over a period of two days. In some embodiments, the infusion is repeated every <NUM> hours over a period of four days. In some embodiments, the infusion is repeated every <NUM> hours over a period of six days. In some embodiments, the infusion is repeated every <NUM> hours over a period of eight days. In some embodiments, the infusion is repeated every <NUM> hours over a period of ten days. In some embodiments, the infusion is repeated every <NUM> hours over a period of <NUM> days. In some embodiments, the infusion is repeated every <NUM> hours over a period of <NUM> days. In some embodiments, the infusion is repeated every <NUM> hours over a period of <NUM> days or more.

In some embodiments, the pharmaceutical composition comprises <NUM> to <NUM> of cefepime. In some embodiments, the pharmaceutical composition comprises <NUM> to <NUM> of cefepime. In some embodiments, the pharmaceutical composition comprises <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, or <NUM> of cefepime. In some embodiments, the pharmaceutical composition comprises <NUM> of cefepime. In some embodiments, the pharmaceutical composition comprises <NUM> of cefepime. In some embodiments, the pharmaceutical composition comprises <NUM> of cefepime. In some embodiments, the pharmaceutical composition comprises <NUM> of cefepime. In some embodiments, the pharmaceutical composition comprises <NUM> of cefepime. In some embodiments, the pharmaceutical composition comprises <NUM> of cefepime. In some embodiments, the pharmaceutical composition comprises <NUM> of cefepime. In some embodiments, the pharmaceutical composition comprises <NUM> of cefepime.

In some embodiments, the daily dose of cefepime is <NUM> to <NUM>. In some embodiments, the daily dose of cefepime is <NUM>, <NUM>, <NUM>, mg, <NUM>, mg, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, or <NUM>. In some embodiments, the daily dose of cefepime is <NUM>. In some embodiments, the daily dose of cefepime is <NUM>. In some embodiments, the daily dose of cefepime is <NUM>. In some embodiments, the daily dose of cefepime is <NUM>. In some embodiments, the daily dose of cefepime is <NUM>. In some embodiments, the daily dose of cefepime is <NUM>. In some embodiments, the daily dose of cefepime is <NUM>. In some embodiments, the daily dose of cefepime is <NUM>. In some embodiments, the daily dose of cefepime is <NUM>.

In some embodiments, the pharmaceutical composition comprises <NUM> to <NUM> of Compound <NUM>. In some embodiments, the pharmaceutical composition comprises <NUM> to <NUM> of Compound <NUM>. In some embodiments, the pharmaceutical composition comprises <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, or <NUM> of Compound <NUM>. In some embodiments, the pharmaceutical composition comprises <NUM> of Compound <NUM>. In some embodiments, the pharmaceutical composition comprises <NUM> of Compound <NUM>. In some embodiments, the pharmaceutical composition comprises <NUM> of Compound <NUM>. In some embodiments, the pharmaceutical composition comprises <NUM> of Compound <NUM>. In some embodiments, the pharmaceutical composition comprises <NUM> of Compound <NUM>. In some embodiments, the pharmaceutical composition comprises <NUM> of Compound <NUM>. In some embodiments, the pharmaceutical composition comprises <NUM> of Compound <NUM>. In some embodiments, the pharmaceutical composition comprises <NUM> of Compound <NUM>. In some embodiments, the pharmaceutical composition comprises <NUM> of Compound <NUM>. In some embodiments, the pharmaceutical composition comprises <NUM> of Compound <NUM>. In some embodiments, the pharmaceutical composition comprises <NUM> of Compound <NUM>. In some embodiments, the pharmaceutical composition comprises <NUM> of Compound <NUM>. In some embodiments, the pharmaceutical composition comprises <NUM> of Compound <NUM>. In some embodiments, the pharmaceutical composition comprises <NUM> of Compound <NUM>. In some embodiments, the pharmaceutical composition comprises <NUM> of Compound <NUM>. In some embodiments, the pharmaceutical composition comprises <NUM> of Compound <NUM>. In some embodiments, the pharmaceutical composition comprises <NUM> of Compound <NUM>. In some embodiments, the pharmaceutical composition comprises <NUM> of Compound <NUM>. In some embodiments, the pharmaceutical composition comprises <NUM> of Compound <NUM>. In some embodiments, the pharmaceutical composition comprises <NUM> of Compound <NUM>. In some embodiments, the pharmaceutical composition comprises <NUM> of Compound <NUM>.

In some embodiments, the pharmaceutical composition comprises <NUM> to <NUM> of Compound <NUM> and <NUM> to <NUM> of cefepime. In certain embodiments, the pharmaceutical composition comprises <NUM> to <NUM> of Compound <NUM> and <NUM> to <NUM> of cefepime. In some embodiments, the pharmaceutical composition comprises <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, or <NUM> of Compound <NUM> or <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, or <NUM> of Compound <NUM>. In certain embodiments, the pharmaceutical composition comprises <NUM> of Compound <NUM> and <NUM> of cefepime. In some embodiments, the pharmaceutical composition comprises <NUM> of Compound <NUM> and <NUM> of cefepime. In certain embodiments, the pharmaceutical composition comprises <NUM> of Compound <NUM> and <NUM> of cefepime. In certain embodiments, the pharmaceutical composition comprises <NUM> of Compound <NUM> and <NUM> of cefepime. In some embodiments, the pharmaceutical composition comprises <NUM> of Compound <NUM> and <NUM> of cefepime. In certain embodiments, the pharmaceutical composition comprises <NUM> of Compound <NUM> and <NUM> of cefepime. In certain embodiments, the pharmaceutical composition comprises <NUM> of Compound <NUM> and <NUM> of cefepime. In some embodiments, the pharmaceutical composition comprises <NUM> of Compound <NUM> and <NUM> of cefepime. In certain embodiments, the pharmaceutical composition comprises <NUM> of Compound <NUM> and <NUM> of cefepime.

In some embodiments, the daily dose of Compound <NUM> is about <NUM> to <NUM>. In some embodiments, the daily dose of Compound <NUM> is about <NUM>, <NUM>, <NUM>, <NUM>, mg, <NUM>, mg, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, or <NUM>. In some embodiments, the daily dose of compound <NUM> is <NUM>. In some embodiments, the daily dose of compound <NUM> is <NUM>. In some embodiments, the daily dose of compound <NUM> is <NUM>. In some embodiments, the daily dose of compound <NUM> is <NUM>. In some embodiments, the daily dose of compound <NUM> is <NUM>. In some embodiments, the daily dose of compound <NUM> is <NUM>. In some embodiments, the daily dose of compound <NUM> is <NUM>. In some embodiments, the daily dose of compound <NUM> is <NUM>. In some embodiments, the daily dose of compound <NUM> is <NUM>. In some embodiments, the daily dose of compound <NUM> is <NUM>. In some embodiments, the daily dose of compound <NUM> is <NUM>. In some embodiments, the daily dose of compound <NUM> is <NUM>. In some embodiments, the daily dose of compound <NUM> is <NUM>. In some embodiments, the daily dose of compound <NUM> is <NUM>. In some embodiments, the daily dose of compound <NUM> is <NUM>. In some embodiments, the daily dose of compound <NUM> is <NUM>.

In some embodiments, the pharmaceutical composition is stored in a sterile container. Compound <NUM> dihydrochloride is co-administered with cefepime.

In some embodiments, Compound <NUM> dihydrochloride and cefepime are provided in separate containers. In some embodiments, Compound <NUM> dihydrochloride and cefepime are provided in a single container. In some embodiments, the container is an IV bag. In some embodiments, the container is a glass bottle. In some embodiments, the container is an amber glass bottle.

In some embodiments, the (R)-<NUM>-(<NUM>-(trans-<NUM>-(<NUM>-aminoethylamino)cyclohexyl)acetamido)-<NUM>-hydroxy-<NUM>,<NUM>-dihydro-<NUM>-benzo[e][<NUM>,<NUM>]oxaborinine-<NUM>-carboxylic acid dihydrochloride and the cefepime are formulated in separate containers. (R)-<NUM>-(<NUM>-(trans-<NUM>-(<NUM>-aminoethylamino)cyclohexyl)acetamido)-<NUM>-hydroxy-<NUM>,<NUM>-dihydro-<NUM>-benzo[e][<NUM>,<NUM>]oxaborinine-<NUM>-carboxylic acid dihydrochloride is in the form of a crystalline form as disclosed herein. In some embodiments, the (R)-<NUM>-(<NUM>-(trans-<NUM>-(<NUM>-aminoethylamino)cyclohexyl)acetamido)-<NUM>-hydroxy-<NUM>,<NUM>-dihydro-<NUM>-benzo[e][<NUM>,<NUM>]oxaborinine-<NUM>-carboxylic acid dihydrochloride is formulated in a first container. In some embodiments, the (R)-<NUM>-(<NUM>-(trans-<NUM>-(<NUM>-aminoethylamino)cyclohexyl)acetamido)-<NUM>-hydroxy-<NUM>,<NUM>-dihydro-<NUM>-benzo[e][<NUM>,<NUM>]oxaborinine-<NUM>-carboxylic acid dihydrochloride is in the form of a first powder for reconstitution. In some embodiments, the cefepime is formulated in a second container. In some embodiments, the cefepime is in the form of a second powder for reconstitution.

Also described herein is a method of administering a pharmaceutical composition, wherein the method comprises:.

In some embodiments of a method of administering a pharmaceutical composition, each of the aqueous carrier is independently water for injection, <NUM>% sodium chloride injection, <NUM>% dextrose injection, <NUM>% dextrose injection, sodium lactate injection, <NUM>% dextrose and <NUM>% sodium chloride injection, lactated Ringers and <NUM>% dextrose injection, sodium chloride/sodium acetate/sodium gluconate/potassium chloride/magnesium chloride injection, sodium chloride/potassium acetate/magnesium acetate in <NUM>% dextrose injection, or any combinations thereof. In some embodiments of a pharmaceutical composition, the composition further comprises a pharmaceutically acceptable excipient. In some embodiments of a pharmaceutical composition, the pharmaceutically acceptable excipient is an amino acid or a monosaccharide derivative. In some embodiments of a pharmaceutical composition, the pharmaceutically acceptable excipient is L-arginine. In some embodiments of a pharmaceutical composition, the pharmaceutically acceptable excipient is meglumine.

In some embodiments, the (R)-<NUM>-(<NUM>-(trans-<NUM>-(<NUM>-aminoethylamino)cyclohexyl)acetamido)-<NUM>-hydroxy-<NUM>,<NUM>-dihydro-<NUM>-benzo[e][<NUM>,<NUM>]oxaborinine-<NUM>-carboxylic acid dihydrochloride and the cefepime are formulated in a single container. In some embodiments, (R)-<NUM>-(<NUM>-(trans-<NUM>-(<NUM>-aminoethylamino)cyclohexyl)acetamido)-<NUM>-hydroxy-<NUM>,<NUM>-dihydro-<NUM>-benzo[e][<NUM>,<NUM>]oxaborinine-<NUM>-carboxylic acid dihydrochloride is in the form of a crystalline form as disclosed herein. In some embodiments of a method of administering a pharmaceutical composition, the (R)-<NUM>-(<NUM>-(trans-<NUM>-(<NUM>-aminoethylamino)cyclohexyl)acetamido)-<NUM>-hydroxy-<NUM>,<NUM>-dihydro-<NUM>-benzo[e][<NUM>,<NUM>]oxaborinine-<NUM>-carboxylic acid dihydrochloride and the cefepime are in the form of a powder for reconstitution. In some embodiments of a method of administering a pharmaceutical composition, the method comprises:.

In some embodiments of a method of administering a pharmaceutical composition, the aqueous carrier is water for injection, <NUM>% sodium chloride injection, <NUM>% dextrose injection, <NUM>% dextrose injection, sodium lactate injection, <NUM>% dextrose and <NUM>% sodium chloride injection, lactated Ringers and <NUM>% dextrose injection, sodium chloride/sodium acetate/sodium gluconate/potassium chloride/magnesium chloride injection, sodium chloride/potassium acetate/magnesium acetate in <NUM>% dextrose injection, or any combinations thereof. In some embodiments of a pharmaceutical composition, the composition further comprises a pharmaceutically acceptable excipient. In some embodiments of a pharmaceutical composition, the pharmaceutically acceptable excipient is an amino acid or a monosaccharide derivative. In some embodiments of a pharmaceutical composition, the pharmaceutically acceptable excipient is L-arginine. In some embodiments of a pharmaceutical composition, the pharmaceutically acceptable excipient is meglumine.

Also described herein are methods for inhibiting bacterial growth, by, e.g., reducing bacterial resistance to a β-lactam antibiotic, such methods comprising contacting a bacterial cell culture, or a bacterially infected cell culture, tissue, or organism, with Compound <NUM> dihydrochloride. The bacteria to be inhibited by administration of Compound <NUM> dihydrochloride may be bacteria that are resistant to beta-lactam antibiotics. The term "resistant" is well-understood by those of ordinary skill in the art (see, e g <NPL>), <NPL>)).

These methods are useful for inhibiting bacterial growth in a variety of contexts. Compound <NUM> dihydrochloride may be administered to an experimental cell culture in vitro to prevent the growth of beta-lactam resistant bacteria. Compound <NUM> dihydrochloride may be administered to a mammal, including a human to prevent the growth of beta-lactam resistant bacteria in vivo. The method may comprise administering a therapeutically effective amount of Compound <NUM> dihydrochloride for a therapeutically effective period of time to a mammal, including a human. Preferably, the Compound <NUM> dihydrochloride is administered in the form of a pharmaceutical composition as described above. In some embodiments, cefepime is co-administered with the Compound <NUM> dihydrochloride.

Also described herein are methods of treating a bacterial infection, which method comprises administering to a subject a pharmaceutical composition comprising Compound <NUM> dihydrochloride and a pharmaceutically acceptable excipient as described above. In some embodiments, the bacterial infection is an upper or lower respiratory tract infection, a urinary tract infection, an intra-abdominal infection, or a skin infection.

In some embodiments, the infection that is treated or prevented comprises a bacteria that includes Elizabethkingia meningoseptica, Pseudomonas aeruginosa, Pseudomonas fluorescens, Pseudomonas acidovorans, Pseudomonas alcaligenes, Pseudomonas putida, Stenotrophomonas maltophilia, Burkholderia cepacia, Aeromonas hydrophilia, Escherichia coli, Citrobacter freundii, Salmonella typhimurium, Salmonella typhi, Salmonella paratyphi, Salmonella enteritidis, Shigella dysenteriae, Shigella flexneri, Shigella sonnei, Enterobacter cloacae, Enterobacter aerogenes, Klebsiella pneumoniae, Klebsiella oxytoca, Serratia marcescens, Francisella tularensis, Morganella morganii, Proteus mirabilis, Proteus vulgaris, Providencia alcahfaciens, Providencia rettgeri, Providencia stuartii, Acinetobacter baumannii, Acinetobacter calcoaceticus, Acinetobacter haemolyticus, Yersinia enterocolitica, Yersinia pestis, Yersinia pseudotuberculosis, Yersinia intermedia, Bordetella pertussis, Bordetella parapertussis, Bordetella bronchiseptica, Haemophilus influenzae, Haemophilus parainfluenzae, Haemophilus haemolyticus, Haemophilus parahaemolyticus, Haemophilus ducreyi, Pasteurella multocida, Pasteurella haemolytica, Branhamella catarrhalis, Helicobacter pylori, Campylobacter fetus, Campylobacter jejuni, Campylobacter coli, Borrelia burgdorferi, Vibrio cholerae, Vibrio parahaemolyticus, Legionella pneumophila, Listeria monocytogenes, Neisseria gonorrhoeae, Neisseria meningitidis, Kingella, Moraxella, Gardnerella vaginalis, Bacteroides fragilis, Bacteroides distasonis, Bacteroides 3452A homology group, Bacteroides vulgatus, Bacteroides ovalus, Bacteroides thetaiotaomicron, Bacteroides uniformis, Bacteroides eggerthii, Bacteroides splanchnicus, Clostridium difficile, Mycobacterium tuberculosis, Mycobacterium avium, Mycobacterium intracellulare, Mycobacterium leprae, Corynebacterium diphtheriae, Corynebacterium ulcerans, Streptococcus pneumoniae, Streptococcus agalactiae, Streptococcus pyogenes, Enterococcus faecalis, Enterococcus faecium, Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus saprophyticus, Staphylococcus intermedius, Staphylococcus hyicus subsp. hyicus, Staphylococcus haemolyticus, Staphylococcus hominis, or Staphylococcus saccharolyticus.

In some embodiments, the infection that is treated or prevented comprises a bacteria that includes Elizabethkingia meningoseptica , Pseudomonas aeruginosa, Pseudomonas fluorescens, Stenotrophomonas maltophilia, Escherichia coli, Citrobacter freundii, Salmonella typhimurium, Salmonella typhi, Salmonella paratyphi, Salmonella enteritidis, Shigella dysenteriae, Shigella flexneri, Shigella sonnei, Enterobacter cloacae, Enterobacter aerogenes, Klebsiella pneumoniae, Klebsiella oxytoca, Serratia marcescens, Acinetobacter calcoaceticus, Acinetobacter haemolyticus, Yersinia enterocolitica, Yersinia pestis, Yersinia pseudotuberculosis, Yersinia intermedia, Haemophilus influenzae, Haemophilus parainfluenzae, Haemophilus haemolyticus, Haemophilus parahaemolyticus, Helicobacter pylori, Campylobacter fetus, Campylobacter jejuni, Campylobacter coli, Vibrio cholerae, Vibrio parahaemolyticus, Legionella pneumophila, Listeria monocytogenes, Neisseria gonorrhoeae, Neisseria meningitidis, Moraxella, Bacteroides fragilis, Bacteroides vulgatus, Bacteroides ovalus, Bacteroides thetaiotaomicron, Bacteroides uniformis, Bacteroides eggerthii, or Bacteroides splanchnicus.

Also described herein is a method of treating bacterial infections by administration of Compound <NUM> dihydrochloride with cefepime. In certain embodiments, the bacterial infection is complicated intra-abdominal infection, complicated urinary tract infection (including pyelonephritis), pneumonia, uncomplicated urinary tract infections, or uncomplicated skin and skin structure infection. In certain embodiments, the bacterial infection is complicated intra-abdominal infection, or complicated urinary tract infection (including pyelonephritis). In certain embodiments, the bacterial infection is complicated intra-abdominal infection. In certain embodiments, the bacterial infection is complicated urinary tract infection (including pyelonephritis).

Also described herein is a method of treating febrile neutropenic patients by administration of Compound <NUM> dihydrochloride in combination with cefepime.

Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of embodiments described herein, certain preferred methods, devices, and materials are now described.

As used herein and in the appended claims, the singular forms "a," "an," and "the" include plural reference unless the context clearly dictates otherwise. Thus, for example, reference to "an excipient" is a reference to one or more excipients and equivalents thereof known to those skilled in the art, and so forth.

The term "about" is used to indicate that a value includes the standard level of error for the device or method being employed to determine the value.

The use of the term "or" in the claims is used to mean "and/or" unless explicitly indicated to refer to alternatives only or the alternatives are mutually exclusive, although the disclosure supports a definition that refers to only alternatives and to "and/or.

The terms "comprise," "have" and "include" are open-ended linking verbs. Any forms or tenses of one or more of these verbs, such as "comprises," "comprising," "has," "having," "includes" and "including," are also open-ended. For example, any method that "comprises," "has" or "includes" one or more steps is not limited to possessing only those one or more steps and also covers other unlisted steps.

"Optional" or "optionally" may be taken to mean that the subsequently described structure, event or circumstance may or may not occur, and that the description includes instances where the events occurs and instances where it does not.

As used herein, the term "therapeutic" means an agent utilized to treat, combat, ameliorate, prevent or improve an unwanted condition or disease of a patient.

"Administering" when used in conjunction with a therapeutic means to administer a therapeutic systemically or locally, as directly into or onto a target tissue, or to administer a therapeutic to a patient whereby the therapeutic positively impacts the tissue to which it is targeted. "Administering" a pharmaceutical composition may be accomplished by injection, topical administration, and oral administration or by other methods alone or in combination with other known techniques.

The term "animal" as used herein includes, but is not limited to, humans and non-human vertebrates such as wild, domestic and farm animals. As used herein, the terms "patient," "subject" and "individual" are intended to include living organisms in which certain conditions as described herein can occur. Examples include humans, monkeys, cows, sheep, goats, dogs, cats, mice, rats, and transgenic species thereof. In a preferred embodiment, the patient is a primate. In certain embodiments, the primate or subject is a human. In certain instances, the human is an adult. In certain instances, the human is child. In further instances, the human is <NUM> years of age or younger. In certain instances, the human is elderly. In other instances, the human is <NUM> years of age or older. Other examples of subjects include experimental animals such as mice, rats, dogs, cats, goats, sheep, pigs, and cows.

By "pharmaceutically acceptable," it is meant the carrier, diluent or excipient must be compatible with the other ingredients of the composition and not deleterious to the recipient thereof.

The term "pharmaceutical composition" means a composition comprising at least one active ingredient, such as Compound <NUM>, whereby the composition is amenable to investigation for a specified, efficacious outcome in a mammal (for example, without limitation, a human). Those of ordinary skill in the art will understand and appreciate the techniques appropriate for determining whether an active ingredient has a desired efficacious outcome based upon the needs of the artisan.

A "therapeutically effective amount" or "effective amount" as used herein refers to the amount of active compound or pharmaceutical agent that elicits a biological or medicinal response in a tissue, system, animal, individual or human that is being sought by a researcher, veterinarian, medical doctor or other clinician, which includes one or more of the following: (<NUM>) preventing the disease; for example, preventing a disease, condition or disorder in an individual that may be predisposed to the disease, condition or disorder but does not yet experience or display the pathology or symptomatology of the disease, (<NUM>) inhibiting the disease; for example, inhibiting a disease, condition or disorder in an individual that is experiencing or displaying the pathology or symptomatology of the disease, condition or disorder (i.e., arresting further development of the pathology and/or symptomatology), and (<NUM>) ameliorating the disease; for example, ameliorating a disease, condition or disorder in an individual that is experiencing or displaying the pathology or symptomatology of the disease, condition or disorder (i.e., reversing the pathology and/or symptomatology).

The terms "treat," "treated," "treatment," or "treating" as used herein refers to both therapeutic treatment in some embodiments and prophylactic or preventative measures in other embodiments, wherein the object is to prevent or slow (lessen) an undesired physiological condition, disorder or disease, or to obtain beneficial or desired clinical results. For the purposes described herein, beneficial or desired clinical results include, but are not limited to, alleviation of symptoms; diminishment of the extent of the condition, disorder or disease; stabilization (i.e., not worsening) of the state of the condition, disorder or disease; delay in onset or slowing of the progression of the condition, disorder or disease; amelioration of the condition, disorder or disease state; and remission (whether partial or total), whether detectable or undetectable, or enhancement or improvement of the condition, disorder or disease. Treatment includes eliciting a clinically significant response without excessive levels of side effects. Treatment also includes prolonging survival as compared to expected survival if not receiving treatment. A prophylactic benefit of treatment includes prevention of a condition, retarding the progress of a condition, stabilization of a condition, or decreasing the likelihood of occurrence of a condition. As used herein, "treat," "treated," "treatment," or "treating" includes prophylaxis in some embodiments.

The term "substantially the same as" as used herein, refers to a powder x-ray diffraction pattern or differential scanning calorimetry pattern that is non-identical to those depicted herein, but that falls within the limits of experimental error, when considered by one of ordinary skill in the art.

In the following examples, crystalline forms <NUM>, <NUM>, <NUM>, <NUM>, and <NUM> are included as reference forms.

XRPD patterns were obtained using the high-throughput XRPD (HT-XRPD) set-up. The plates were mounted on a Bruker GADDS diffractometer equipped with a Hi-Star area detector. The XRPD platform was calibrated using Silver Behenate for the long d-spacings and Corundum for the short d-spacings.

Data collection was carried out at room temperature using monochromatic CuKα radiation in the 2θ region between <NUM>° and <NUM>°, which is the most distinctive part of the XRPD pattern. The diffraction pattern of each well was collected in two <NUM>θ ranges (<NUM>°≤ <NUM><NUM>° for the first frame, and <NUM>°≤ 2θ <NUM>° for the second) with an exposure time of <NUM> for each frame. No background subtraction or curve smoothing was applied to the XRPD patterns.

The carrier material used during XRPD analysis was transparent to X-rays and contributed only slightly to the background.

The powder data were collected on D8 Advance diffractometer using Cu Kα1 radiation (<NUM>Å) with germanium monochromator at Room Temperature. The data were collected in <NUM>θ alone (detector scan) mode from <NUM> to <NUM>° <NUM>θ with <NUM>° <NUM>θ steps, <NUM> per step on solid state LynxEye detector. The sample was measured in <NUM> long glass capillary with <NUM> outer diameter.

For variable temperature experiments the ANSYCO HT chamber was used, installed within the D8 Advance system diffractometer (Bruker) designed with Brag-Brentano geometry and equipped with LynxEye solid state detector. The radiation used for collecting the data was CuKα1 (λ = <NUM>Å) monochromatized by germanium crystal. The material was placed on a fixed sample holder that was mounted inside the chamber.

VT-XRPD: The temperature variation rate was <NUM>/min. The patterns were collected in the range <NUM> - <NUM>2θ, with a step of <NUM>2θ and measuring time per step of <NUM> sec. The data collection time, per temperature, was <NUM>*<NUM>.

Melting properties were obtained from DSC thermograms, recorded with a heat flux DSC822e instrument (Mettler-Toledo GmbH, Switzerland). The DSC822e was calibrated for temperature and enthalpy with a small piece of indium (melting point at <NUM> C; ΔHf = <NUM> J. Samples were sealed in standard <NUM>µl aluminum pans, pin-holed and heated in the DSC from <NUM> to <NUM>, at a heating rate of <NUM> min-<NUM>. Dry N2 gas, at a flow rate of <NUM> min-<NUM> was used to purge the DSC equipment during measurement.

Mass loss due to solvent or water loss from the crystals was determined by TGA/SDTA (TGA: thermogravimetric analysis; SDTA: single differential thermal analysis). Monitoring the sample weight, during heating in a TGA/SDTA851e instrument (Mettler-Toledo GmbH, Switzerland), resulted in a weight vs. temperature curve. The TGA/SDTA851e was calibrated for temperature with indium and aluminum. Samples were weighed into <NUM>µl aluminum crucibles and sealed. The seals were pin-holed and the crucibles heated in the TGA from <NUM> to <NUM> or <NUM> (depending on the experiment) at a heating rate of <NUM> min-<NUM>. Dry N2 gas was used for purging.

The gases evolved from the TGA samples were analyzed by a mass spectrometer Omnistar GSD <NUM> T2 (Pfeiffer Vacuum GmbH, Germany). The latter is a quadrupole mass spectrometer, which analyzes masses in the range of <NUM>-<NUM> amu.

Differences in hygroscopicity (moisture uptake) of the various forms of a solid material provided a measure of their relative stability at increasing relative humidity. Moisture sorption isothermal profiles of small samples were obtained using a DVS-<NUM> system from Surface Measurement Systems (London, UK); this instrument is suitable for use with as little as a few milligrams of sample, with an accuracy of <NUM>µg.

The following humidity profile was applied: The relative humidity was cycled from <NUM>% to <NUM>% (sorption), back to <NUM>% (desorption) and to <NUM>%, at a constant temperature of <NUM>. Steps consisted of <NUM>% RH (between <NUM>% and <NUM>% RH), <NUM>% RH (between 0and <NUM>% RH) and <NUM>% (between <NUM>% and <NUM>% RH). Weight equilibration per step was set with a minimum holding time of <NUM> hour. At the end of the DVS experiments, the recovered solid material was measured by HT-XRPD.

To a solution of tert-butyl <NUM>-((R)-<NUM>-(<NUM>-((1r,4R)-<NUM>-((tert-butoxycarbonyl)(<NUM>-((tert-butoxycarbonyl)amino)ethyl)amino)cyclohexyl)acetamido)-<NUM>-((3aS,<NUM>,<NUM>,7aR)-3a,<NUM>,<NUM>-trimethylhexahydro-<NUM>,<NUM>-methanobenzo[d][<NUM>,<NUM>,<NUM>]dioxaborol-<NUM>-yl)ethyl)-<NUM>-methoxybenzoate (Structure <NUM>, <NUM>, <NUM> mmol) in <NUM>,<NUM> dioxane (<NUM>) was added hydrochloric acid (<NUM>, <NUM> in water). The resulting mixture was heated to reflux and stirred at this temperature for <NUM> minutes. The resulting mixture was cooled to room temperature over <NUM> minutes then extracted with methyl tert-butyl ether (3x200 mL). The aqueous phase was concentrated under vacuum (<NUM>-<NUM> mmHg and <NUM>-<NUM> external bath temperature, wherein <NUM> mmHg = <NUM> Pa). The residue was diluted to a total weight of <NUM> with water. To this solution was added isopropanol (<NUM>) over approximately <NUM> minutes. This solution was seeded with Compound <NUM>-(HCl)<NUM> crystals (<NUM>). To this mixture was added isopropanol (<NUM>) over approximately <NUM> minutes. The resulting slurry was stirred for approximately <NUM> hours then filtered under vacuum, the solid was washed with isopropanol (2x200 mL) then methyl tert-butyl ether (<NUM>). The solid was further dried under high vacuum to give the title compound (structure <NUM>) (<NUM>) as a white crystalline solid as a mixture of Form <NUM> and Form <NUM>.

In order to design a re-crystallization process for Compound <NUM>-dihydrochloride salt from water as solvent and IPA as anti-solvent, the solubility of the API in these combinations were determined. Suspensions in various water/IPA mixtures were prepared and incubated for <NUM> hours at <NUM>°, <NUM>°, <NUM>° and <NUM>. Upon completion of the incubation time the solids were separated from the mother liquors by centrifugation and filtration and the concentration of Compound <NUM> in solution was determined by HPLC analysis. The remaining solids were analyzed by HT-XRPD. The results of the solubility determination as a function of anti-solvent content and temperature is presented in Table <NUM> and in <FIG>. Table <NUM>: Solubility values of Compound <NUM>-dihydrochloride in mg/ml in IPA/water mixtures at four temperatures, <NUM>, <NUM>, <NUM> and <NUM>.

To Compound <NUM>-dihydrochloride from example <NUM>, (<NUM>, Sample A) was added <NUM> of isopropanol/water (<NUM>/<NUM> v/v). The mixture was stirred at <NUM> to <NUM> until a clear solution was obtained. The solution was filtered through a membrane filter (<NUM> micron). To the filtrate was added isopropanol (<NUM>) followed by seed crystals (<NUM>). To this mixture was added isopropanol (<NUM>) over <NUM> hour. The resulting mixture was stirred for <NUM> hours then filtered under vacuum. The wet cake was deliquored under vacuum for <NUM> hour <NUM> minutes. The solid was washed with isopropanol (<NUM>) then methyl tert-butyl ether (<NUM>). The solid was dried under a flow of ambient air over <NUM> hours <NUM> minutes. Recovered <NUM> of Compound <NUM>-dihydrochloride monohydrate (Sample B). Chloride by ion chromatography <NUM>% w/w. Water content by Karl Fischer analysis= <NUM>% w/w.

Table <NUM> shows the impurity profile of Compound <NUM>-dihydrochloride (Sample A) and crystalline Compound <NUM>-dihydrochloride (Sample B) as assessed by HPLC method described in Table <NUM>.

Crystalline Compound <NUM>-dihydrochloride (from example 2B) was analyzed by HR-XRPD (<FIG>). Based on the single crystal data available for the monohydrate (<FIG>) it was concluded that the crystalline form represents the monohydrate of the dihydrochloride salt of Compound <NUM>. It was called Form <NUM>. Table <NUM> lists the peaks positions and their intensities for Form <NUM>.

Analysis of Compound <NUM>-dihydrochloride monohydrate by TG/MS (<FIG>) showed that a <NUM>% mass loss was recorded between <NUM>° and <NUM>. This mass is attributed to the loss of <NUM> molecule of water per molecule of Compound <NUM>-dihydrochloride. This result confirms the XRPD analysis that the material consisted of the monohydrated form. The compound decomposed at temperatures over <NUM>.

<FIG> shows the DSC trace of Form <NUM> for a programmed increase in temperature from <NUM> to <NUM>. The DSC trace shows an endothermic event with an onset temperature of approximately <NUM>. This event corresponds to the onset of dehydration of the monohydrate. This dehydration process is followed by the decomposition of the compound at about <NUM> signaled by a large endothermic event. It is concluded from this experiment that Form <NUM> is stable to elevated temperature, up to approximately <NUM>.

Compound <NUM>-dihydrochloride monohydrate was analyzed by dynamic vapor sorption (<FIG>). The material was subjected to an isothermal desorption/sorption profile starting at <NUM>% - <NUM>% - <NUM>% - <NUM>% relative humidity levels (step <NUM>% at low relative humidity (RH)) at <NUM>.

The temperature was maintained constant at approximately <NUM> for the duration of the experiment. The curve with the diamond markers shows the change in mass of a sample of Form <NUM> when exposed to increasing relative humidity level (<NUM>% RH to <NUM>%). It can be seen that the mass uptake is rather small and that at high humidity level Compound <NUM>-dihydrochloride remained as Form <NUM>. This is confirmed by the mass stability and the VH-XRPD experiment conducted (see <FIG>).

The curve with the square markers shows the change in mass of a sample of Form <NUM> when subjected to a series of decreasing relative humidities from <NUM>%RH to <NUM>%RH. It can be seen that the sample mass does not change significantly between approximately <NUM>%RH to <NUM>%RH. Upon subsequent exposure to humidity values of <NUM>, <NUM>, and <NUM>%RH, the sample loses approximately <NUM>% of its mass which corresponds to the loss of a single molecule of water per Compound <NUM>-dihydrochloride salt. A VH-XRPD experiment confirmed that the loss of water was accompanied by a transformation to the anhydrous form (Form <NUM>) (see <FIG>).

The curve with the triangle markers shows the change in the mass of Form <NUM> produced by the dehydration of crystalline Form <NUM>, when exposed to increasing relative humidity (<NUM> %RH to <NUM> %RH). There is a slight increase in mass up to <NUM>%RH, followed by a sharp increase in mass when Form <NUM> was exposed to <NUM>%RH. A VH-XRPD experiment showed that Form <NUM> converts to Form <NUM> at %RH values in the <NUM>-<NUM>%RH range (see <FIG>). These observations demonstrate that Form <NUM> undergoes significant changes in moisture content and physical form at relative humidity values that are representative of values specified in pharmaceutical manufacturing facilities.

To confirm the hypothesis of a solid form change between <NUM>% and <NUM>% relative humidity levels a variable humidity XRPD (VH-XRPD) analysis was performed. The sample was subjected to a humidity profile of <NUM> - <NUM> - <NUM> - <NUM> - <NUM> - <NUM> - <NUM> - <NUM>% relative humidity levels at <NUM>. At each step, two diffractograms were recorded. The first diffractogram was recorded when the target humidity was reached and the second after the sample had equilibrated for <NUM> hour at each humidity level. When a phase change occurred and if the conversion rate was faster than the scan time, the diffractogram showed peaks of the starting form at the start (low 2θ angles) and peaks of the converted form at the end of the diffractogram (higher 2θ angles). The most intense peaks of Form <NUM> (dihydrochloride monohydrate) are in the range between <NUM> and <NUM>° 2θ.

The stacked diffractograms recorded at different relative humidity levels are shown in <FIG>. The measured RH level and the kind of ongoing RH variation step (sorption or desorption) is found on the right hand side. The material displays a clear change in diffractogram indicating a moisture sorption induced phase transition. Upon desorption, the material was converted between <NUM>% and <NUM>% relative humidity to Form <NUM>, the anhydrous form. Upon sorption, Form <NUM> converted back to Form <NUM> between <NUM>% and <NUM>% relative humidity. DVS analysis showed a significant moisture sorption in the range between <NUM>% and <NUM>% RH. The VH-XRPD results did not raise evidence for additional form changes at these values.

From the variable-humidity X-ray powder diffraction experiments performed on Form <NUM> a second crystalline form of Compound <NUM>-dihydrochloride was discovered. This form was designated Form <NUM>. For further characterization of Form <NUM>, some material was prepared by vacuum drying Form <NUM> at <NUM> for <NUM> hours. <FIG> shows the XRPD diffractogram of Form <NUM>. Analysis of a solid sample of Form <NUM> by TGA demonstrated the anhydrous nature of this solid. The thermogravimetric and the thermogram obtained by DSC analysis showed the absence of any thermal events before decomposition at <NUM> and the absence of any weight loss on drying (<FIG>). Table <NUM> lists the peaks positions and their intensities for Form <NUM>.

The VH-XRPD and DVS analysis performed on the starting materials suggested that there is a dynamic and reversible relationship between the anhydrous Form <NUM> and the monohydrate Form <NUM>. The conversion rate of Form <NUM> to Form <NUM> as a function of the relative humidity value was investigated by a isothermal moisture sorption experiment at fixed relative humidity values. In brief, a solid sample of Form <NUM> was prepared by drying Form <NUM>. This sample was placed in the DVS machine and incubated for <NUM> hour at the indicated humidity level while the weight change was recorded. The results are presented in <FIG>. The conversion rate depended on the relative humidity level. The highest rate was achieved at <NUM>% RH whereas the lowest rate was achieved at <NUM>% RH. The right panel shows the linear relationship between the applied humidify level and the conversion rate.

Compound <NUM>-dihydrochloride was physically characterized by X-ray powder diffraction, thermal analysis, and dynamic vapor sorption techniques. Loss-on-drying studies performed by TG/MS analysis demonstrated that the material consisted of the monohydrated crystalline solid form of Compound <NUM>-dihydrochloride. This particular form was designated Form <NUM>. The material showed decomposition at temperatures above <NUM>. Characterization by DVS analysis showed a dehydration/hydration pattern between <NUM>% and <NUM>% relative humidity values at <NUM> with a clear hysteresis. The presence of the hysteresis suggested a phase change from a monohydrate to an anhydrous crystalline form. This phase transition could be confirmed by a variable X-ray powder diffraction study that showed conversion of Form <NUM> (monohydrate) to Form <NUM> (anhydrous).

Physical characterization of Compound <NUM>-dihydrochloride has confirmed the existence of two crystalline forms. The main form is the monohydrate designated Form <NUM> and the other form is an anhydrous form, Form <NUM>. Dynamic vapor sorption studies have shown that under ambient conditions the monohydrated form of Compound <NUM>-dihydrochloride is the most stable form. Only at low relative humidity levels or drying under elevated temperature and/or under vacuum the anhydrous form is observed in this study. The interconversion between the two solid forms is fast and reversible.

A known amount of Compound <NUM> was weighed directly into <NUM> vials and <NUM> of each phosphate buffer was added; the solutions obtained were kept at controlled temperature (<NUM>) and magnetically stirred up to <NUM> hours. Aliquots of obtained suspensions or solutions were centrifuged (<NUM> rpm <NUM> minutes) and withdrawn at two predefined time points (<NUM> hours and <NUM> hours) appropriately diluted and analyzed by HPLC-UV using the analytical method parameters summarized in Table <NUM>.

Solubility data of Compound <NUM>-dihydrochloride monohydrate and the pH values are reported in Table <NUM>.

A physical stability study of Compound <NUM>-dihydrochloride (mixture of Form <NUM> and Form <NUM>) was conducted. A solid sample of Compound <NUM>-dihydrochloride was incubated under four relative humidity and temperature conditions: <NUM>/ambient RH, <NUM>/<NUM>% RH, <NUM>/<NUM>% RH and <NUM>/<NUM>% RH. At regular intervals, the compound was analyzed by HR-XRPD and TGMS. After three days exposure, the initial crystal form of Compound <NUM>-dihydrochloride showed full conversion to Form <NUM> (monohydrate) at all four conditions tested. No significant changes in the solid form were observed upon incubation for extended period of times (at least up to <NUM> month).

The water content of each sample was determined by TGMS analysis and is presented in <FIG>. After <NUM> days exposure, the water content increased from <NUM>% to values between <NUM> and <NUM>% (corresponding to <NUM> or <NUM> molecule of water per molecule of Compound <NUM>-dihydrochloride. The water content is in line with the monohydrate nature of Compound <NUM>-dihydrochloride Form <NUM>. The TGMS analyses recorded for the <NUM> weeks and <NUM>-month incubation is also along the <NUM>% of water (<NUM> molecule of water per molecule of API).

This data shows that Form <NUM> is physically stable and non-hygroscopic under the tested conditions. No changes in the crystalline phase were observed for the samples incubated for <NUM> weeks. Furthermore, no significant changes in the water content were recorded upon exposure to various conditions and times.

Generation of Compound <NUM>-dihydrochloride amorphous powder.

Approximately <NUM> of Compound <NUM>-dihydrochloride was weighed into nine separate <NUM> glass vials, and nine selected solvent systems were added to these vials in steps until a clear solution was obtained. In several solvent systems (<NUM>-propanol-water, <NUM>,<NUM>-dioxane/water, tetrahydrofuran/water, ethanol/water, acetone/water, and acetonitrile/water; all in a <NUM>/<NUM> v/v ratio) the API did not dissolve; therefore, these experiments were discarded. The solutions (derived from methanol/water, <NUM>/<NUM> v/v ratio; dimethylsulfoxide/water, <NUM>/<NUM> v/v ratio; and water) were frozen in liquid nitrogen followed by drying using Crys Alpha <NUM>-<NUM> LO for <NUM> hours. An extra drying at <NUM> and <NUM> mbar was applied for <NUM> hours. The resulting solids were analyzed by HT-XRPD. From one solvent, water, an amorphous solid was obtained. These conditions were scaled-up to produce <NUM> of amorphous solid for the polymorph screen.

The design of a comprehensive polymorph screen is characterized by utilizing different crystallization techniques combined with a variety of solvents and solvent mixtures. The diversity of a screen will depend on the solubility profile of the API. Certain crystallization modes such as equilibration of suspensions require an intermediate solubility, whereas, anti-solvent crystallization relies on solvents with good solubility and solvents with a close to zero solubility.

The polymorph screen comprised of the following crystallization modes:.

The solids obtained from the various crystallization experiments were analyzed by High Throughput XRPD (HT-XRPD). If relevant, the mother liquors were allowed to evaporate completely, and the remaining solids were analyzed by HT-XRPD. Subsequently, all solids were exposed to accelerated aging conditions for <NUM> hours at <NUM> and <NUM>%RH (AAC).

From the polymorph screen, four novel diffractograms were identified in addition to the two known forms of Compound <NUM>-dihydrochloride, Form <NUM> and Form <NUM>. Form <NUM> was the most abundant solid form found in this polymorph screen. The novel diffractograms were designated Forms <NUM>, <NUM>, <NUM> and <NUM>. All these forms were found with low frequency and were poorly crystalline. They all were physically unstable as conversion to Form <NUM> was observed upon exposure to short term stress conditions (<NUM> days at <NUM> and <NUM>%RH). The HT-XRPD patterns of the new Forms <NUM>, <NUM>, <NUM> and <NUM> are shown in <FIG>, <FIG>, <FIG>, and <FIG>, respectively. Table <NUM> provides the summary of the experimental conditions under which these forms were obtained.

All above mentioned forms except for Form <NUM> were further analyzed by DSC, TGMS, and HPLC. A short description of the analytical characterization is presented below.

Form <NUM> was obtained as a pure crystalline phase in a single experiment. No further analysis could be done on this form due to the limited physical stability. This form was found in the solid obtained from iso-amyl alcohol after being dried under ambient condition. Upon drying under vacuum, conversion to Form <NUM> occurred. Also, upon exposure to short term stress conditions Form <NUM> transformed into Form <NUM>.

Form <NUM> was found in a few crystallization experiments performed in TFE, <NUM>,<NUM>-dioxane and IPA/water. The thermal analysis confirmed that Form <NUM> is a mixed solvated/hydrated form. Based on the different solvent/water contents determined for several Form <NUM> samples, we assumed that Form <NUM> is a class of isostructural mixed hydrate/solvates.

A mixed hydrated/iso-amyl alcohol solvated form was identified from the solvent equilibration experiments performed in iso-amyl alcohol. This new solid form was designated Form <NUM> which seemed to be physically stable upon drying under vacuum; however, upon exposure to AAC conversion to Form <NUM> was observed.

Form <NUM> was predominately found during the hydrate screen experiments performed in IPA/water mixtures but also it was identified in a few solvent equilibration experiments performed in different solvents, such as <NUM>,<NUM>-dimethoxyethane, DMSO/DCM and DMF/anisole. Form <NUM> was mainly found from the solids dried under vacuum; however, conversion to Form <NUM> was observed after AAC. The analytical characterization suggested that Form <NUM> is a mixed hydrated/solvated form. Based on the different solvents that can be incorporated to the structure of Form <NUM> we assumed that this form is clustering a class of isostructural mixed hydrate/solvates containing different organic solvent molecules.

Solvent equilibration experiments were performed in several water/organic solvents mixtures to investigate the formation of other hydrated forms. Suspensions of amorphous Compound <NUM>-dihydrochloride were prepared in <NUM> water/solvent mixtures with different water activities. The suspensions were stirred for <NUM> days at <NUM>, <NUM>, <NUM> and <NUM>.

After completion of the equilibration time, the remaining solids were analyzed by HT-XRPD dry under ambient conditions and dried under vacuum. The mother liquors were analyzed by Karl Fisher titration for water content determination. Three experiments did not show solids after the equilibration conditions. The solutions from these experiments were evaporated under vacuum (<NUM> mBar) and the obtained dried solids analyzed by HT-XRPD. All the solids were then exposed to <NUM> and <NUM>% RH for two days.

From most of the crystallization conditions, the monohydrate Form <NUM> had crystallized.

From IPA/water mixtures, Form <NUM>, Form <NUM> and mixtures of those with Form <NUM> were identified at low water contents (<<NUM>%). At water activity values above <NUM>, the monohydrated Form <NUM> was identified in all cases. Based on the analytical characterization, Form <NUM> and <NUM> seemed to be mixed hydrated/solvated forms which upon exposure to short term stress conditions converted to Form <NUM>.

The solvent equilibration experiments performed in ethyl acetate/water showed differences in the solid form assessed to the solids dried under ambient and under vacuum. Form <NUM> was identified in most of the solids dried under ambient conditions. Upon drying under vacuum, mixtures of Form <NUM> and <NUM> were identified. Form <NUM> was identified in mixture with Form <NUM> at water activity values below <NUM> at temperatures above <NUM>. The presence of Form <NUM> was observed when elevated temperatures and high-water contents were used but also at temperatures below <NUM> and low water contents.

Similar trend was observed for the experiments performed in acetonitrile/water. All the solids dried under ambient conditions were attributed to Form <NUM>. Upon drying the solids under vacuum, in some cases partial conversion to Form <NUM> was observed. This observation was noticed for the solvent equilibration experiments performed at temperatures above <NUM>. The experiments performed below <NUM> did not show any solid form conversion upon drying.

The physical stability study performed for <NUM> days at <NUM>/<NUM>%RH led to the identification of Form <NUM>. All new powder patterns detected in this study showed conversion to the initial Form <NUM>.

The stability of aqueous solutions of Compound <NUM>-dihydrochloride in combination with meropenem at pH <NUM> vs of Compound <NUM>-dihydrochloride in combination with cefepime at pH <NUM> after <NUM> hours was studied. The results are shown in Table <NUM>.

The results shown in examples 7A-7D show that the cefepime/Compound <NUM> combination is more efficacious than the meropenem/Compound <NUM> combination against a variety of organisms.

Elizabethkingia meningoseptica is a Gram negative rod that expresses <NUM> chromosomal metallo-beta-lactamase (both carbapenemases) and a class A serine cephalosporinase CME (AAC, <NUM>, <NUM>-<NUM>). The MIC testing results (N=<NUM> strains) are shown in Table <NUM>.

Stenotrophomonas maltophilia is a Gram negative bacillus. It is an opportunistic MDR pathogen. It has been linked to infections with high morbidity and mortality in severely immunocompromised and debilitated individuals. The MIC testing results are shown in Table <NUM>.

<NUM> isolates non-susceptible to meropenem (MIC values to meropenem ><NUM>µg/mL). The MIC testing results are shown in Table <NUM>.

<NUM> isolates non-susceptible to cefepime (MIC values to cefepime ><NUM>µg/mL). The MIC testing results are shown in Table <NUM>.

Example <NUM> describes four Phase <NUM> clinical studies designed to evaluate the safety, tolerability and PK of various dose-levels of Compound <NUM> when administered as single or multiple doses either alone or in combination with cefepime comprising the following components (and outlined in Table <NUM>):.

The results from in vitro and in vivo nonclinical PK/PD studies, the human PK data collected in the Phase <NUM> Study, and human plasma binding data were used to determine human therapeutic doses required for PK/PD target attainment. In vitro studies have demonstrated that human serum protein binding of Compound <NUM> is <NUM>%, leaving a free fraction of <NUM>%. Table <NUM> summarizes the calculated values for the total AUC<NUM>-<NUM>, the corresponding free fraction (fAUC<NUM>-<NUM>) and the fAUC<NUM>-<NUM>/MIC ratio estimates based on the established CLSI breakpoint for cefepime of <NUM>/L, when administered at a <NUM> dose tid.

The fAUC<NUM>-<NUM>/MIC ratios (based on an MIC value of <NUM>/L) for all doses ≥<NUM> q8 hr exceed the PK/PD target required to achieve bacterial stasis for the isolates studied to date. Similarly, the fAUC<NUM>-<NUM>/MIC values for doses of approximately <NUM> q8 hr (interpolated from shown data) exceed the fAUC<NUM>-<NUM>/MIC ratios required to achieve <NUM> log of bacterial killing. Based on these data, a dose of <NUM> q8hr provides approximately three times the drug exposure required for PK/PD target attainment for bacterial stasis.

<NUM> subjects overall divided between two study parts as follows:.

No exemptions will be granted for Inclusion/Exclusion criteria. The following inclusion criteria must be met for a subject to be eligible for inclusion in the study:.

A subject who meets any of the following exclusion criteria will not be eligible for inclusion in the study:.

Compound <NUM> was provided in glass vials containing <NUM> of a freeze dried solid. Compound <NUM> was reconstituted at the site with sterile water for injection. The site obtained the following products from a commercial pharmacy or wholesaler:.

This is a Phase <NUM>, randomized, single-center, drug-drug interaction study in healthy adult male and female volunteers. The study consisted of two independent parts. Each part enrolled separate groups of subjects. Eligible subjects were allowed to participate in either Part <NUM> or Part <NUM>, but not both.

Part <NUM> enrolled <NUM> subjects into randomized, single dose assessment of five treatments with a sequential <NUM>-period crossover (Part 1A) and <NUM>-period crossover (Part 1B) design. Subjects who had not met any halting rules in Part 1A were eligible for re-randomization into Part 1B.

Part <NUM> enrolled <NUM> subjects into a randomized, double-blind assessment of three treatments in a parallel group design.

In view of the available nonclinical data, and data from single and multiple dose administration in humans, further development of Compound <NUM> in combination with a β-lactam antibiotic was warranted. This study provided an assessment of the safety and pharmacokinetics of Compound <NUM> combined with cefepime with and without metronidazole in a multi-period single dose cross-over design in Part <NUM>, and of the safety, tolerability, and pharmacokinetics (PK) of multiple doses of Compound <NUM> administered in combination with cefepime in Part <NUM>.

Compound <NUM> was administered as an intravenous (IV) infusion over <NUM> hours.

Due to the study design, the low risk of clinically significant toxicity at anticipated exposure levels, and the absence of any potential therapeutic benefit, healthy subjects were chosen for this study population. Moreover, use of healthy subjects as opposed to patients allowed a clearer interpretation of the study results, as there was no confounding factors resulting from changes in disease state and/or concomitant medications.

All treatments in Part 1A were administered as a single <NUM>-hour IV infusion and included the treatments in Table <NUM>. All subjects received all three treatments. Dosing occured on Day <NUM>, Day <NUM>, and Day <NUM>. The sequence of administration was randomized.

The Part 1A treatment descriptions are in Table <NUM>.

Part 1B includeed two treatments, which were administered according to the treatment sequence specified in the randomization code. Dosing occured on Day <NUM> and Day <NUM>. Treatments are described in Table <NUM>.

In Part <NUM>, subjects were randomized in a <NUM>:<NUM>:<NUM> ratio to one of the three treatments in Table <NUM>.

Treatment was administered every <NUM> hours (q8h) as <NUM>-hour IV infusion for <NUM> days. Subjects received a total of <NUM> doses with a single dose administered on Day <NUM>.

There was no significant difference in the pharmacokinetic parameters of AUC(<NUM>-inf) and Cmax for Compound <NUM> or cefepime upon co-administration of the two drugs relative to administration of either drug alone.

There was no meaningful pharmacokinetic interaction between metronidazole and cefepime/Compound <NUM> relative to administration of metronidazole alone or the cefepime/Compound <NUM> combination without metronidazole.

Both single and repeat doses of study medication were well-tolerated. There were no clinically significant vital sign, ECG or laboratory abnormalities. All adverse events were mild in intensity.

Subjects with infections at multiple anatomic sites caused by resistant pathogens including lower respiratory tract infections, complicated intra-abdominal infections (cIAI) and complicated urinary tract infections (cUTI), such as Pyelonephritis, are administered a combination of cefepime/ Compound <NUM> as follows in Table <NUM>.

Claim 1:
A crystalline form of (R)-<NUM>-(<NUM>-(trans-<NUM>-(<NUM>-aminoethylamino)cyclohexyl)acetamido)-<NUM>-hydroxy-<NUM>,<NUM>-dihydro-<NUM>-benzo[e][<NUM>,<NUM>]oxaborinine-<NUM>-carboxylic acid dihydrochloride:
<CHM>
wherein the crystalline form is anhydrous; and
wherein the crystalline form has an X-ray powder diffraction (XRPD) pattern that includes at least three characteristic peaks selected from peaks at:
<NUM>°±<NUM>° 2θ, <NUM>°±<NUM>° 2θ, <NUM>°±<NUM>° 2θ, <NUM>°±<NUM>° 2θ, <NUM>°±<NUM>° 2θ, <NUM>°±<NUM>° 2θ, <NUM>°±<NUM>° 2θ, <NUM>°±<NUM>° 2θ, <NUM>°±<NUM>° 2θ, <NUM>°±<NUM>° 2θ, <NUM>°±<NUM>° 2θ, and <NUM>°±<NUM>° 2θ.