JP7490329B2 - Pharmaceutical Composition for Treating Crohn's Disease - Google Patents

Description

Applicable under Article 30, paragraph 2 of the Patent Act Posted on the website (May 21, 2016) https://ddw.scientificposters.com/epsAbstractDDW.cfm?id=1

Article 30, paragraph 2 of the Patent Act applies. Posting on the website (since April 22, 2016) Digestive Disease Week 2016 Mobile app

Presentation at the meeting on the application of Article 30, Paragraph 2 of the Patent Act (May 23, 2016) Digestive Disease Week 2016, San Diego, CA, USA

Application of Article 30, Paragraph 2 of the Patent Act Publication (April 19, 2016) Gastroenterology (2016), Vol. 150, Issue 4

Application of Article 30, Paragraph 2 of the Patent Act Posted on the website (April 16, 2016) http://www.gastrojournal.org/article/S0016-5085(16)32732-9/pdf

The present invention relates to a pharmaceutical composition for treating Crohn's disease, comprising an anti-fractalkine antibody.

Fractalkine (also called "FKN") is a membrane-bound chemokine that is expressed on the surface of vascular endothelial cells in response to inflammatory stimuli such as LPS, TNF-α, and IL-1. Cells expressing the FKN receptor CX3CR1 bind to membrane-bound FKN without the intervention of selectins or integrins, causing strong cell adhesion. In addition, secreted FKN shed from membrane-bound FKN exhibits cell migration activity for NK cells, T cells, and monocytes that have CX3CR1.

Expression of FKN is induced on the surface of vascular endothelial cells by proinflammatory cytokines. Elevated expression of FKN and accumulation of CX3CR1 ⁺ cytotoxic effector lymphocytes and macrophages have been reported in patients with Crohn's disease (also called "CD").

To date, FKN has been considered as a promising therapeutic target for inflammatory bowel diseases (also referred to as "IBD") such as ulcerative colitis (also referred to as "UC") and CD (Non-Patent Document 1). It has also been suggested that anti-fractalkine antibodies that inhibit the interaction between FKN and CX3CR1 may be able to treat IBD, including Crohn's disease (Patent Document 1). Thus, it has been suggested that antibodies that can bind to FKN and inhibit the action of FKN are effective in treating Crohn's disease.

The present applicant has previously reported several mouse anti-human fractalkine (hFKN) monoclonal antibodies (clones 1F3-1, 3A5-2, 1F3, 1G1, 2B2, 3D5, 3H7, 6D1, 7F6, and 5H7-6), and in particular clone 3A5-2 has been humanized and named H3-2L4 due to its high neutralizing activity, binding affinity, and species cross-reactivity against hFKN (Patent Document 2, the entirety of which is incorporated by reference).

Ann. NY Acad. Sci. 2009;1173:350-356

WO2006/046739 WO2011/052799

An object of the present invention is to provide a pharmaceutical composition comprising an anti-fractalkine antibody (also referred to herein as "anti-FKN antibody") that brings about a therapeutically effective improvement in Crohn's disease after administration to a human subject.
Another object of the present invention is to provide pharmaceutical compositions comprising anti-FKN antibodies for use in bringing about therapeutically effective improvement in Crohn's disease.

The present invention encompasses the following embodiments.
[1] A pharmaceutical composition for treating Crohn's disease, comprising:
The pharmaceutical composition comprises an anti-fractalkine antibody and a pharma- ceutically acceptable excipient;
The anti-fractalkine antibody is
the heavy chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 13 (QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYYIHWVKQAPGQGLEWIGWIYPGDGSPKFNERFKGRTTLTADKSTNTAYMLLSSLRSEDTAVYFCATGPTDGDYFDYWGQGTTVTVSS);
the light chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 14 (DIQMTQSPSSLSASVGDRVTITCRASGNIHNFLAWYQQKPGKAPKLLIYNEKTLADGVPSRFSGSGSGTDYTLTISSLQPEDFATYFCQQFWSTPYTFGGGTKVEIK);
a constant region of a human IgG2 isotype; and an Fc region of the constant region of the human IgG2 isotype comprising the mutations V234A and G237A,
The pharmaceutical composition is used so that the anti-fractalkine antibody is intravenously administered to a human in a single dose of at least 10 mg/kg of human body weight.
The pharmaceutical composition.

[2] The pharmaceutical composition according to [1],
The pharmaceutical composition is characterized in that it is used so that the anti-fractalkine antibody is intravenously administered to a human in an amount of 10 to 15 mg/kg of human body weight per administration.
The pharmaceutical composition.

[3] The pharmaceutical composition according to [1],
When administered intravenously in a single dose to humans, the anti-fractalkine antibody has a mean _Cmax of 21 to 25 μg/mL per 1 mg of the anti-fractalkine antibody/kg of human body weight.
The pharmaceutical composition.

[4] The pharmaceutical composition according to [1],
When the anti-fractalkine antibody is administered intravenously in a single dose of 10 mg/kg of human body weight to a human, the mean C _max of the anti-fractalkine antibody is within a numerical range of 80% to 125% of 2.4×10 ² μg/mL.
The pharmaceutical composition.

[5] The pharmaceutical composition according to [1],
When the anti-fractalkine antibody is administered intravenously once at 10 mg/kg of human body weight to a human, the mean AUC _(0-t) of the anti-fractalkine antibody is within a numerical range of 80% to 125% of 7.0×10 ⁴ μg·h/mL.
The pharmaceutical composition.

[6] The pharmaceutical composition according to [1],
When the anti-fractalkine antibody is administered intravenously once at 10 mg/kg of human body weight to a human, the mean AUC _{(0-336 h)} of the anti-fractalkine antibody is within a numerical range of 80% to 125% of 3.8×10 ⁴ μg·h/mL.
The pharmaceutical composition.

[7] The pharmaceutical composition according to [1],
Administered intravenously in multiple doses at intervals ranging from once per week to once every two weeks;
The pharmaceutical composition.

[8] The pharmaceutical composition according to any one of [1] to [7],
The anti-fractalkine antibody is administered intravenously so that the mean trough concentration of the anti-fractalkine antibody is 80 μg / mL or more.
The pharmaceutical composition.

[9] A pharmaceutical composition for treating Crohn's disease, comprising:
The pharmaceutical composition comprises a therapeutically effective amount of an anti-fractalkine antibody and a pharma- ceutical acceptable excipient;
The anti-fractalkine antibody is
the heavy chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 13 (QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYYIHWVKQAPGQGLEWIGWIYPGDGSPKFNERFKGRTTLTADKSTNTAYMLLSSLRSEDTAVYFCATGPTDGDYFDYWGQGTTVTVSS);
the light chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 14 (DIQMTQSPSSLSASVGDRVTITCRASGNIHNFLAWYQQKPGKAPKLLIYNEKTLADGVPSRFSGSGSGTDYTLTISSLQPEDFATYFCQQFWSTPYTFGGGTKVEIK);
a constant region of a human IgG2 isotype; and an Fc region of the constant region of the human IgG2 isotype comprising the mutations V234A and G237A,
The therapeutically effective amount is such that, when a pharmaceutical composition containing the anti-fractalkine antibody is administered intravenously in a single dose, the mean _Cmax of the anti-fractalkine antibody per 1 mg of the anti-fractalkine antibody/kg of human body weight is 21 to 25 μg/mL.
The pharmaceutical composition.

[10] A therapeutic agent for Crohn's disease comprising an anti-fractalkine antibody,
The method is characterized in that an anti-fractalkine antibody is intravenously administered to a human at a dose of 10 to 15 mg/kg of human body weight per administration at an administration interval of once a week to once every two weeks;
The anti-fractalkine antibody is
the heavy chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 13 (QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYYIHWVKQAPGQGLEWIGWIYPGDGSPKFNERFKGRTTLTADKSTNTAYMLLSSLRSEDTAVYFCATGPTDGDYFDYWGQGTTVTVSS);
the light chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 14 (DIQMTQSPSSLSASVGDRVTITCRASGNIHNFLAWYQQKPGKAPKLLIYNEKTLADGVPSRFSGSGSGTDYTLTISSLQPEDFATYFCQQFWSTPYTFGGGTKVEIK);
a constant region of a human IgG2 isotype; and an Fc region of the constant region of the human IgG2 isotype comprising the mutations V234A and G237A.
The drug for treating Crohn's disease.

Inventions that combine one or more of the above-mentioned aspects of the present invention in any manner are also included within the scope of the present invention.

In accordance with the present invention, there is provided a pharmaceutical composition comprising an anti-FKN antibody that provides a therapeutically effective improvement in Crohn's disease following administration to a human subject. The pharmaceutical composition of the present invention can be used to provide pharmacokinetic parameters of the anti-FKN antibody useful for exerting a therapeutic effect against Crohn's disease.

FIG. 1 shows the mean serum H3-2L4 concentrations over time following a single intravenous dose of antibody H3-2L4 (0.04 mg/kg, 0.2 mg/kg, 1 mg/kg, 3 mg/kg, 6 mg/kg and 10 mg/kg dose groups, respectively). FIG. 2 shows the mean serum H3-2L4 concentrations over time following multiple intravenous doses of antibody H3-2L4 (2 mg/kg, 5 mg/kg, 10 mg/kg and 15 mg/kg dose groups). FIG. 3 shows the mean serum total FKN concentrations over time following multiple intravenous doses of antibody H3-2L4 (2 mg/kg, 5 mg/kg, 10 mg/kg and 15 mg/kg dose groups). FIG. 4 shows the binding occupancy of antibody H3-2L4 against mFKN over time based on simulations using the QSS model. FIG. 5 shows the ratio of the amount of cFKN to the amount of cFKN before the start of administration over time, based on a simulation using a QSS model. FIG. 6 shows CRP over time up to 12 weeks after administration in seven subjects in Cohort 3 (10 mg/kg group).

1. SUMMARY AND DEFINITIONS OF THEINVENTION The present invention relates to pharmaceutical compositions that provide therapeutically effective improvement in Crohn's disease.

In one embodiment of the present invention, "therapeutically effective improvement in Crohn's disease" may mean showing therapeutically effective improvement in one or more of the evaluation criteria for Crohn's disease established in the field. Such evaluation criteria may include, but are not limited to, Crohn's disease activity index (also referred to as "CDAI"), which shows the disease activity of inflammatory bowel disease symptoms such as diarrhea, abdominal pain, general condition, extraintestinal complications, perianal abscess, fistula, fever, and weight loss as an activity index, The International Organization for the study of Inflammatory Bowel Disease score (also referred to as "IOIBDscore"), Dutch activity index (also referred to as "Dutch-AI"), etc. Another example is detecting the level of C-reactive protein (also called "CRP") in the patient's serum. The severity of disease according to the CDAI is classified as follows: less than 150 indicates remission, 150-219 indicates mild disease, 220-450 indicates moderate disease, and over 450 indicates severe disease activity.

In one embodiment of the present invention, if the pre-treatment CDAI is 220 or more, and the post-treatment CDAI shows a reduction of 70 or more, preferably 100 or more, from the pre-treatment CDAI as a baseline, or preferably shows remission with a post-treatment CDAI of less than 150, this indicates that there has been a "therapeutically effective improvement in Crohn's disease."

In one aspect, the present invention relates to a pharmaceutical composition for treating Crohn's disease that provides a therapeutically effective serum concentration of anti-FKN antibodies after a single or multiple administration to a human subject.

In one embodiment of the present invention, the "therapeutically effective serum anti-FKN antibody concentration" is a serum concentration of anti-FKN antibody of 20 μg/mL or more, 30 μg/mL or more, 40 μg/mL or more, 50 μg/mL or more, 60 μg/mL or more, 70 μg/mL or more, or 80 μg/mL or more. In another embodiment of the present invention, the "therapeutically effective serum anti-FKN antibody concentration" is a mean trough concentration of anti-FKN antibody of 20 μg/mL or more, 30 μg/mL or more, 40 μg/mL or more, 50 μg/mL or more, 60 μg/mL or more, 70 μg/mL or more, or 80 μg/mL or more. In a preferred embodiment, the "therapeutically effective serum anti-FKN antibody concentration" is a mean trough concentration of anti-FKN antibody of 70 μg/mL or more or 80 μg/mL or more. In a more preferred embodiment, a "therapeutically effective serum anti-FKN antibody concentration" is a mean trough concentration of anti-FKN antibodies of 80 μg/mL or greater.

Thus, in one aspect, the pharmaceutical composition of the present invention is formulated to provide a serum concentration of anti-FKN antibody of 20 μg/mL or more, 30 μg/mL or more, 40 μg/mL or more, 50 μg/mL or more, 60 μg/mL or more, 70 μg/mL or more, or 80 μg/mL or more. In a preferred embodiment, the pharmaceutical composition of the present invention is formulated to provide a serum concentration of anti-FKN antibody of 70 μg/mL or more. In another preferred embodiment of the present invention, the pharmaceutical composition of the present invention is formulated to provide a serum concentration of anti-FKN antibody of 80 μg/mL or more.

In another aspect, the present invention relates to a pharmaceutical composition comprising an anti-FKN antibody, which is used to provide a therapeutically effective serum anti-FKN antibody concentration after single or multiple administration to a human subject. The pharmaceutical composition of the present invention may be, but is not limited to, typically in the form of an injectable formulation prepared for intravenous administration. In one embodiment, the pharmaceutical composition of the present invention is used to provide a mean trough concentration of the anti-FKN antibody of 20 μg/mL or more, 30 μg/mL or more, 40 μg/mL or more, 50 μg/mL or more, 60 μg/mL or more, 70 μg/mL or more, or 80 μg/mL or more. In a preferred embodiment, the pharmaceutical composition of the present invention is used to provide a mean trough concentration of the anti-FKN antibody of 70 μg/mL or more, or 80 μg/mL or more.

In the present invention, the pharmacokinetic characteristics of the concentration-time curve, such as the maximum observed serum concentration (C _max ), the time to reach C _max (T _max ), and the area under the serum concentration versus time curve (AUC), are examined by statistical methods well established in the field of pharmacokinetics. When the ratio of the population means of the above-mentioned evaluation parameters between the test preparation and the reference preparation is 0.80 to 1.25, they are generally considered to be bioequivalent.

In the present invention, the average values of pharmacokinetic parameters such as _Cmax and AUC can be calculated by any of the following methods: geometric mean, arithmetic mean, and median. Unless otherwise specified, the average _Cmax , average AUC, average trough concentration, and average tFNK concentration are shown as arithmetic mean values in this specification. Even if the target average value is calculated by a method different from that of this specification, it is intended that the average value falls within the numerical range described in the claims of the present invention when the average value calculated according to the method of this specification is within the numerical range described in the claims of the present invention.

In the present invention, "one time" or "per time" in relation to the dosage of anti-FKN antibody refers to the absolute amount of anti-FKN antibody per administration. Therefore, for example, when it is described as "10 mg/kg human body weight per administration," it means that an amount of anti-FKN antibody calculated by multiplying 10 mg by human body weight (kg) is administered by one set of the pharmaceutical composition of the present invention, or two or more sets used simultaneously.

In the present invention, with regard to the administration interval of the anti-FKN antibody, for example, "an administration interval of once a week to once every two weeks" means that the administration interval from the nth administration (n is an integer) to the n+1th administration is one to two weeks, and the administration interval between the nth and n+1th administrations and the administration interval between the n+1st and n+2th administrations may be different. For example, "an administration interval of once a week to once every two weeks" naturally includes the case where the administration interval between the first and second administrations is one week, and the administration interval between the second and third administrations is two weeks.

As used herein, the term "human" or "human subject" is intended to mean a healthy adult male and typically any human exhibiting clinical signs and symptoms of Crohn's disease or any disease or disorder that may result in Crohn's disease. In the present invention, a "human" or "human subject" is preferably a Crohn's disease patient who has had insufficient, persistent or intolerant therapy with at least one standard treatment (corticosteroids, immunomodulators and/or anti-TNF antibodies).
2. Anti-FKN antibody

In the present invention, when referring to an anti-FKN antibody, it refers to the humanized anti-human fractalkine antibody H3-2L4 or an antibody functionally equivalent thereto. In the present invention, a "functionally equivalent antibody" refers to an antibody that is equivalent to the antibody H3-2L4 in at least one of, and preferably several of, the binding affinity for human FKN, neutralizing activity, cross-reactivity, and pharmacokinetics in blood.

In the present invention, when referring to anti-FKN antibody, it may include its antigen-binding fragment. Such antigen-binding fragment is not particularly limited as long as it is a functional and structural fragment of anti-FKN antibody, which retains the binding ability to FKN and does not significantly differ in pharmacokinetics in blood from the complete antibody. Examples of antigen-binding fragment of antibody include, but are not limited to, Fab, Fab', F(ab') ₂ , Fv, single chain (ScFv), their mutants, fusion proteins containing antibody portions, and other modified structures of immunoglobulin molecules containing antigen recognition sites.

In one embodiment, the anti-FKN antibody of the present invention can be any antibody comprising the following CDR sequences:
(a) CDR-H1 comprising the amino acid sequence of SEQ ID NO:15 (NYYIH);
(b) CDR-H2 comprising the amino acid sequence of SEQ ID NO: 16 (WIYPGDGSPKFNERFKG);
(c) CDR-H3 comprising the amino acid sequence of SEQ ID NO: 17 (GPTDGDYFDY);
(d) CDR-L1 comprising the amino acid sequence of SEQ ID NO: 18 (RASGNIHNFLA);
(e) CDR-L2 comprising the amino acid sequence of SEQ ID NO: 19 (NEKTLAD); and (f) CDR-L3 comprising the amino acid sequence of SEQ ID NO: 20 (QQFWSTPYT).

In another embodiment, the anti-FKN antibody comprises a heavy chain and a light chain, and the heavy chain variable region of the antibody is represented by SEQ ID NO: 21 (QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYYIHWVKQAPGQGLEWIGWIYPGDGSPKFNERFKGRTTLTADKSTNTAYMLLSSLRSDDTAVYFCATGPTDGDYFDYWGQGTTVTVSS), SEQ ID NO: 13 (QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYYIHWVKQAPGQGLEWIGWIYPGDGSPKFNERFKGRTTLTADKSTNTAYMLLSSLRSED TAVYFCATGPTDGDYFDYWGQGTTVTVSS), sequence number 22 (QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYYIHWVRQAPGQGLEWIGWIYPGDGSPKFNERFKGRTTLTRDKSTNTAYMELSSLRSDDTAVYFCATGPTDGDYFDYWGQGTTVTVSS), sequence number 23 (QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYYIHWVRQAPGQGLEWIGWIYPGDGSPKFNERFKGRTTMTDTSTSTAYMELSSLRSEDTAVYFCARGPTDGDY and the light chain variable region of the antibody comprises the amino acid sequence of SEQ ID NO: 25 (DIQMTQSPSSLSASVGDRVTITCRASGNIHNFLAWYQQKPGKAPKFLVYNEKTLADGVPSRFSGSGSGTQYTLTISSLQPEDFATYFCQQF WSTPYTFGGGTKVEIK), SEQ ID NO: 14 (DIQMTQSPSSLSASVGDRVTITCRASGNIHNFLAWYQQKPGKAPKLLIYNEKTLADGVPSRFSGSGSGTDYTLTISSLQPEDFATYFCQQFWSTPYTFGGGTKVEIK), or SEQ ID NO: 26 (DIQMTQSPSSLSASVGDRVTITCRASGNIHNFLAWYQQKPGKAPKLLIYNEKTLADGVPSRFSGSGSGTQYTLTISSLQPEDFATYFCQQFWSTPYTFGGGTKVEIK).

In a preferred embodiment, the anti-FKN antibody may comprise a heavy chain and a light chain, the heavy chain variable region of the antibody may comprise the amino acid sequence of SEQ ID NO: 13 (QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYYIHWVKQAPGQGLEWIGWIYPGDGSPKFNERFKGRTTLTADKSTNTAYMLLSSLRSEDTAVYFCATGPTDGDYFDYWGQGTTVTVSS), and the light chain variable region of the antibody may comprise the amino acid sequence of SEQ ID NO: 14 (DIQMTQSPSSLSASVGDRVTITCRASGNIHNFLAWYQQKPGKAPKLLIYNEKTLADGVPSRFSGSGSGTDYTLTISSLQPEDFATYFCQQFWSTPYTFGGGTKVEIK).

In certain embodiments, the anti-FKN antibody is an antibody that includes a constant region of the human IgG2 isotype.

In a specific embodiment, the anti-FKN antibody is an antibody in which the Fc region of the constant region of the human IgG2 isotype contains the mutations V234A and/or G237A.

In a particularly preferred embodiment of the present invention, the anti-FKN antibody has the amino acid sequence shown in SEQ ID NO: 11 (QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYYIHWVKQAPGQGLEWIGWIYPGDGSPKFNERFKGRTTLTADKSTNTAYMLLSSLRSEDTAVYFCATGPTDGDYFDYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPAAAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTI The antibody H3-2L4 is composed of a heavy chain consisting of SKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK) and a light chain consisting of the amino acid sequence shown in SEQ ID NO: 12 (DIQMTQSPSSLSASVGDRVTITCRASGNIHNFLAWYQQKPGKAPKLLIYNEKTLADGVPSRFSGSGSGTDYTLTISSLQPEDFATYFCQQFWSTPYTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC).

The antibodies of the present invention also include the above-mentioned anti-FKN antibodies that have been appropriately modified (e.g., modified, or partially substituted, added, or deleted from the amino acid sequence of the antibody) while retaining the function of the antibody or adding or improving the function of the antibody. More specifically, the scope of the present invention also includes antibodies in which lysine (Lys) located at the carboxy terminus (C terminus) of the heavy chain has been deleted by an artificial method such as genetic modification in order to reduce the heterogeneity of the antibodies produced by the antibody-producing cells. In addition, the anti-FKN antibodies contained in the pharmaceutical composition of the present invention do not necessarily have to be completely homogeneous, and may contain a mixture of antibodies in which lysine (Lys) located at the carboxy terminus (C terminus) of the heavy chain has been deleted and those in which it has not been deleted, as long as the pharmaceutical composition of the present invention maintains the intended function.

The anti-FKN antibody may be modified, if desired, to alter (a) the three-dimensional structure of the amino acid sequence in the modified region, e.g., a sheet or helix conformation; (b) the charge or hydrophobicity state of the molecule at the target site; or (c) the effect of the modification on maintaining the volume of the side chains, or such modifications may not be clearly observed.

Modification of the anti-FKN antibody may be achieved, for example, by substitution, deletion, addition, etc. of the constituent amino acid residues.

In this specification, the term "amino acid" is used in its broadest sense and includes not only naturally occurring amino acids such as serine (Ser), asparagine (Asn), valine (Val), leucine (Leu), isoleucine (Ile), alanine (Ala), tyrosine (Tyr), glycine (Gly), lysine (Lys), arginine (Arg), histidine (His), aspartic acid (Asp), glutamic acid (Glu), glutamine (Gln), threonine (Thr), cysteine (Cys), methionine (Met), phenylalanine (Phe), tryptophan (Trp), and proline (Pro), but also non-naturally occurring amino acids such as amino acid variants and derivatives. In light of this broad definition, those skilled in the art will understand that amino acids herein include, for example, L-amino acids; D-amino acids; chemically modified amino acids such as amino acid variants and amino acid derivatives; amino acids that are not components of proteins in vivo, such as norleucine, β-alanine, ornithine; and chemically synthesized compounds having the properties of amino acids known to those skilled in the art. Examples of unnatural amino acids include α-methylamino acids (such as α-methylalanine), D-amino acids (such as D-aspartic acid and D-glutamic acid), histidine-like amino acids (such as 2-amino-histidine, β-hydroxy-histidine, homohistidine, α-fluoromethyl-histidine, α-methyl-histidine), amino acids with an extra methylene in the side chain ("homo" amino acids), and amino acids in which the carboxylic acid functional group in the side chain is replaced with a sulfonic acid group (such as cysteic acid).

Naturally occurring amino acid residues can be classified into the following groups, for example, based on common side chain properties:
(1) Hydrophobic: Met, Ala, Val, Leu, Ile;
(2) Neutral hydrophilic: Cys, Ser, Thr;
(3) Acidic: Asp, Glu;
(4) basic: Asn, Gln, His, Lys, Arg;
(5) residues that affect chain orientation: Gly, Pro; and (6) aromatics: Trp, Tyr, Phe.

Non-conservative substitutions in the amino acid sequence constituting the anti-FKN antibody may be made by exchanging an amino acid belonging to one of these groups with an amino acid belonging to another group. More conservative substitutions may be made by exchanging an amino acid belonging to one of these groups with another amino acid of the same group. Similarly, deletions or substitutions in the amino acid sequence may be made as appropriate.
3. Pharmaceutical Compositions and Formulations

The dosage form of the pharmaceutical composition of the present invention is not particularly limited, but is typically a formulation prepared for intravenous administration (e.g., an injectable formulation). The pharmaceutical composition of the present invention can be prepared as an injectable formulation, for example, by dissolving an anti-FKN antibody in, but not limited to, water for injection, saline, or phosphate buffered saline together with a pharma- ceutically acceptable excipient. Pharmaceutically acceptable excipients used in the present invention include, but are not limited to, stabilizers, surfactants, preservatives, and the like. Representative excipients and processes for the manufacture of injectable formulations are known in the art, and are described, for example, in Introduction to Pharmaceutical Dosage Forms, 1985, Ansel, H. C., Lea and Febiger, Philadelphia, Pa. ; Remington's Pharmaceutical Sciences, 1995, Mack Publ. Co., Easton, Pa., which is incorporated herein by reference in its entirety.

In one embodiment of the present invention, the pharmaceutical composition of the present invention is a formulation for intravenous administration, which, when administered intravenously in a single dose to a human, has a mean _Cmax of the anti-fractalkine antibody per 1 mg of the anti-fractalkine antibody/kg of human body weight of 21 to 25 μg/mL.

In another embodiment of the present invention, the pharmaceutical composition of the present invention is a formulation for intravenous administration in which the mean C _max of the anti-fractalkine antibody is a value within the numerical range of 80% to 125% of 2.4 x 10 ² μg/mL when the anti-fractalkine antibody is administered intravenously in a single dose of 10 mg/kg of human body weight to a human.

In another embodiment of the present invention, the pharmaceutical composition of the present invention is a formulation for intravenous administration, which has a mean AUC _(0-t) of the anti-fractalkine antibody within the numerical range of 80% to 125% of 7.0 x 10 ⁴ μg·h/mL when the anti-fractalkine antibody is administered intravenously in a single dose of 10 mg/kg of human body weight to a human.

In another embodiment of the present invention, the pharmaceutical composition of the present invention is a formulation for intravenous administration, in which the mean AUC _{(0-336 h)} of the anti-fractalkine antibody is a value within the numerical range of 80% to 125% of 3.8 x 10 ⁴ μg·h/mL when the anti-fractalkine antibody is administered intravenously in a single dose of 10 mg/kg of human body weight to a human.

In another embodiment of the invention, the pharmaceutical composition of the invention is an intravenous formulation having at least any two of the following characteristics:
- a mean _Cmax of the anti-fractalkine antibody per mg of the anti-fractalkine antibody/kg of human body weight of 21-25 μg/mL when administered as a single intravenous dose to humans;
- the mean C _max of the anti-fractalkine antibody is within the range of 80% to 125% of 2.4 x 10 ² μg/mL when the anti-fractalkine antibody is administered intravenously in a single dose of 10 mg/kg of human body weight to a human;
- when 10 mg/kg of the anti-fractalkine antibody is administered intravenously in a single dose to a human, the mean AUC _(0-t) of the anti-fractalkine antibody is within the numerical range of 80% to 125% of 7.0 x 10 ⁴ μg·h/mL; and - when 10 mg/kg of the anti-fractalkine antibody is administered intravenously in a single dose to a human, the mean AUC _(0-336h) of the anti-fractalkine antibody is within the numerical range of 80% to 125% of 3.8 x 10 ⁴ μg·h/mL.

4. Dosage, Dosage Interval and Dosage Frequency The pharmaceutical composition of the present invention is administered to a human subject at a dosage that will bring about a therapeutically effective improvement in Crohn's disease.Thus, the pharmaceutical composition of the present invention is administered so that the blood concentration or mean trough concentration of anti-FKN antibody is 20 μg/mL or more, 30 μg/mL or more, 40 μg/mL or more, 50 μg/mL or more, 60 μg/mL or more, 70 μg/mL or more, or 80 μg/mL or more.Preferably, the pharmaceutical composition of the present invention is administered multiple times so that the mean trough concentration of anti-FKN antibody is 70 μg/mL or more, or 80 μg/mL or more.

In one embodiment of the present invention, the pharmaceutical composition of the present invention is used to intravenously administer to a human at least 10 mg/kg of human body weight per dose of anti-FKN antibody. In another embodiment of the present invention, the pharmaceutical composition of the present invention is used to intravenously administer to a human at 10 mg/kg to 15 mg/kg of human body weight per dose of anti-FKN antibody. In one embodiment of the present invention, the pharmaceutical composition of the present invention is used to intravenously administer to a human at 10 mg/kg or 15 mg/kg of human body weight per dose of anti-FKN antibody.

The number of doses and the interval between doses of the pharmaceutical composition of the present invention may vary depending on the amount of anti-FKN antibody administered per dose and the route of administration.

In one embodiment of the present invention, the pharmaceutical composition of the present invention is used to intravenously administer multiple doses of at least 10 mg/kg human body weight of anti-FKN antibody to a human subject in need thereof, with a dosing interval of once a week to once every two months. In another embodiment of the present invention, the pharmaceutical composition of the present invention is used to intravenously administer multiple doses of at least 10 mg/kg human body weight of anti-FKN antibody to a human subject in need thereof, with a dosing interval of once a week to once every two months. In another embodiment of the present invention, the pharmaceutical composition of the present invention is used to intravenously administer multiple doses of at least 10 mg/kg human body weight of anti-FKN antibody to a human subject in need thereof, with a dosing interval of once a week to once every two weeks. In a specific embodiment of the present invention, the pharmaceutical composition of the present invention is used to intravenously administer multiple doses of at least 10 mg/kg human body weight of anti-FKN antibody to a human subject in need thereof, with a dosing interval of once every two weeks after two weekly doses.

In another embodiment of the present invention, the pharmaceutical composition of the present invention is used to intravenously administer multiple doses of 10-15 mg/kg human body weight of anti-FKN antibody to a human subject in need thereof at a dosing interval of once a week to once every two months. In another embodiment of the present invention, the pharmaceutical composition of the present invention is used to intravenously administer multiple doses of 10-15 mg/kg human body weight of anti-FKN antibody to a human subject in need thereof at a dosing interval of once a week to once every two months. In another embodiment of the present invention, the pharmaceutical composition of the present invention is used to intravenously administer multiple doses of 10-15 mg/kg human body weight of anti-FKN antibody to a human subject in need thereof at a dosing interval of once a week to once every two weeks. In a specific embodiment of the present invention, the pharmaceutical composition of the present invention is used to intravenously administer 10-15 mg/kg human body weight of anti-FKN antibody to a human subject in need thereof at a dosing interval of once every two weeks after two weekly doses.

In another embodiment of the present invention, the pharmaceutical composition of the present invention is used to intravenously administer multiple doses of 10 mg/kg human body weight or 15 mg/kg human body weight of anti-FKN antibody to a human subject in need thereof at a dosing interval of once a week to once every two months. In another embodiment of the present invention, the pharmaceutical composition of the present invention is used to intravenously administer multiple doses of 10 mg/kg human body weight or 15 mg/kg human body weight of anti-FKN antibody to a human subject in need thereof at a dosing interval of once a week to once every two months. In another embodiment of the present invention, the pharmaceutical composition of the present invention is used to intravenously administer multiple doses of 10 mg/kg human body weight or 15 mg/kg human body weight of anti-FKN antibody to a human subject in need thereof at a dosing interval of once a week to once every two weeks. In a specific embodiment of the present invention, the pharmaceutical composition of the present invention is used to intravenously administer 10 mg/kg human body weight or 15 mg/kg human body weight of anti-FKN antibody to a human subject in need thereof at a dosing interval of once every two weeks after two weekly doses.

The number of administrations of the pharmaceutical composition of the present invention is not particularly limited as long as it brings about a therapeutically effective improvement in Crohn's disease, and may vary depending on the amount of anti-FKN antibody administered per administration, the administration route, and the administration interval.

In one embodiment of the present invention, the pharmaceutical composition of the present invention may be administered intravenously at least 2 times, at least 3 times, at least 4 times, at least 5 times, at least 6 times, at least 7 times, at least 8 times, at least 9 times, at least 10 times, at least 11 times, at least 12 times, at least 13 times, at least 14 times, at least 15 times, at least 16 times, at least 17 times, at least 18 times, at least 19 times, at least 20 times, at least 21 times, at least 22 times, at least 23 times, at least 24 times, at least 25 times, at least 26 times, at least 27 times or more, with administration intervals ranging from once per week to once every two months.

In another embodiment of the invention, the pharmaceutical composition of the invention may be administered intravenously at least 2 times, at least 3 times, at least 4 times, at least 5 times, at least 6 times, at least 7 times, at least 8 times, at least 9 times, at least 10 times, at least 11 times, at least 12 times, at least 13 times, at least 14 times, at least 15 times, at least 16 times, at least 17 times, at least 18 times, at least 19 times, at least 20 times, at least 21 times, at least 22 times, at least 23 times, at least 24 times, at least 25 times, at least 26 times, at least 27 times or more, with administration intervals ranging from once a week to once a month.

In yet another embodiment of the present invention, the pharmaceutical composition of the present invention may be administered intravenously at least 2 times, at least 3 times, at least 4 times, at least 5 times, at least 6 times, at least 7 times, at least 8 times, at least 9 times, at least 10 times, at least 11 times, at least 12 times, at least 13 times, at least 14 times, at least 15 times, at least 16 times, at least 17 times, at least 18 times, at least 19 times, at least 20 times, at least 21 times, at least 22 times, at least 23 times, at least 24 times, at least 25 times, at least 26 times, at least 27 times or more with administration intervals ranging from once per week to once every two weeks.

In certain embodiments of the invention, the pharmaceutical compositions of the invention may be administered intravenously at least 2 times, at least 3 times, at least 4 times, at least 5 times, at least 6 times, at least 7 times, at least 8 times, at least 9 times, at least 10 times, at least 11 times, at least 12 times, at least 13 times, at least 14 times, at least 15 times, at least 16 times, at least 17 times, at least 18 times, at least 19 times, at least 20 times, at least 21 times, at least 22 times, at least 23 times, at least 24 times, at least 25 times, at least 26 times, at least 27 times or more at biweekly dosing intervals following two weekly doses.

In certain embodiments of the present invention, the pharmaceutical composition of the present invention may be administered intravenously at least twice, at least three times, at least four times, at least five times, at least six times, at least seven times, at least eight times, at least nine times, at least ten times, at least eleven times, at least twelve times, at least thirteen times, at least fourteen times, at least fifteen times, at least sixteen times, at least seventeen times, at least eighteen times, at least nineteen times, at least twenty times, at least twenty-one times, at least twenty-two times, at least twenty-three times, at least twenty-four times, at least twenty-five times, at least twenty-six times, at least twenty-seven times, or more, at intervals ranging from once a week to once every two months, such that the mean trough concentration of anti-FKN antibodies is 20 μg/mL or more, 30 μg/mL or more, 40 μg/mL or more, 50 μg/mL or more, 60 μg/mL or more, 70 μg/mL or more, or 80 μg/mL or more.

In yet another embodiment of the present invention, the pharmaceutical composition of the present invention may be administered intravenously at least twice, at least three times, at least four times, at least five times, at least six times, at least seven times, at least eight times, at least nine times, at least ten times, at least eleven times, at least twelve times, at least thirteen times, at least fourteen times, at least fifteen times, at least sixteen times, at least seventeen times, at least eighteen times, at least nineteen times, at least twenty times, at least twenty-one times, at least twenty-two times, at least twenty-three times, at least twenty-four times, at least twenty-five times, at least twenty-six times, at least twenty-seven times, or more, at intervals of once a week to once a month, so that the mean trough concentration of the anti-FKN antibody is 20 μg/mL or more, 30 μg/mL or more, 40 μg/mL or more, 50 μg/mL or more, 60 μg/mL or more, 70 μg/mL or more, or 80 μg/mL or more.

In yet another embodiment of the present invention, the pharmaceutical composition of the present invention may be administered intravenously at least twice, at least three times, at least four times, at least five times, at least six times, at least seven times, at least eight times, at least nine times, at least ten times, at least eleven times, at least twelve times, at least thirteen times, at least fourteen times, at least fifteen times, at least sixteen times, at least seventeen times, at least eighteen times, at least nineteen times, at least twenty times, at least twenty-one times, at least twenty-two times, at least twenty-three times, at least twenty-four times, at least twenty-five times, at least twenty-six times, at least twenty-seven times, or more, at intervals of once a week to once every two weeks, so that the mean trough concentration of the anti-FKN antibody is 20 μg/mL or more, 30 μg/mL or more, 40 μg/mL or more, 50 μg/mL or more, 60 μg/mL or more, 70 μg/mL or more, or 80 μg/mL or more.

In further embodiments of the present invention, the pharmaceutical composition of the present invention may be administered intravenously at least 2 times, at least 3 times, at least 4 times, at least 5 times, at least 6 times, at least 7 times, at least 8 times, at least 9 times, at least 10 times, at least 11 times, at least 12 times, at least 13 times, at least 14 times, at least 15 times, at least 16 times, at least 17 times, at least 18 times, at least 19 times, at least 20 times, at least 21 times, at least 22 times, at least 23 times, at least 24 times, at least 25 times, at least 26 times, at least 27 times or more at biweekly administration intervals after two weekly administrations, so that the mean trough concentration of anti-FKN antibodies is 20 μg/mL or more, 30 μg/mL or more, 40 μg/mL or more, 50 μg/mL or more, 60 μg/mL or more, 70 μg/mL or more, or 80 μg/mL or more.

The following non-limiting examples illustrate certain aspects of the present invention.
In one aspect, the present invention provides a pharmaceutical composition for treating Crohn's disease, the pharmaceutical composition comprising an anti-fractalkine antibody and a pharma- ceutical acceptable excipient;
The fractalkine antibody comprises:
the heavy chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 13 (QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYYIHWVKQAPGQGLEWIGWIYPGDGSPKFNERFKGRTTLTADKSTNTAYMLLSSLRSEDTAVYFCATGPTDGDYFDYWGQGTTVTVSS);
the light chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 14 (DIQMTQSPSSLSASVGDRVTITCRASGNIHNFLAWYQQKPGKAPKLLIYNEKTLADGVPSRFSGSGSGTDYTLTISSLQPEDFATYFCQQFWSTPYTFGGGTKVEIK);
a constant region of a human IgG2 isotype; and an Fc region of the constant region of the human IgG2 isotype comprising the mutations V234A and G237A,
The pharmaceutical composition is characterized in that it is used so that the anti-fractalkine antibody is administered intravenously to a human in a single dose of at least 10 mg/kg of human body weight.

In another aspect, the present invention provides a pharmaceutical composition for treating Crohn's disease, the pharmaceutical composition comprising an anti-fractalkine antibody and a pharma- ceutical acceptable excipient;
The fractalkine antibody comprises:
the heavy chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 13 (QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYYIHWVKQAPGQGLEWIGWIYPGDGSPKFNERFKGRTTLTADKSTNTAYMLLSSLRSEDTAVYFCATGPTDGDYFDYWGQGTTVTVSS);
the light chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 14 (DIQMTQSPSSLSASVGDRVTITCRASGNIHNFLAWYQQKPGKAPKLLIYNEKTLADGVPSRFSGSGSGTDYTLTISSLQPEDFATYFCQQFWSTPYTFGGGTKVEIK);
a constant region of a human IgG2 isotype; and an Fc region of the constant region of the human IgG2 isotype comprising the mutations V234A and G237A,
The pharmaceutical composition is characterized in that it is used so that the anti-fractalkine antibody is administered intravenously to a human at 10 to 15 mg/kg of human body weight per administration.

In yet another aspect, the present invention provides a pharmaceutical composition for treating Crohn's disease, the pharmaceutical composition comprising an anti-fractalkine antibody and a pharma- ceutical acceptable excipient;
The fractalkine antibody comprises:
the heavy chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 13 (QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYYIHWVKQAPGQGLEWIGWIYPGDGSPKFNERFKGRTTLTADKSTNTAYMLLSSLRSEDTAVYFCATGPTDGDYFDYWGQGTTVTVSS);
the light chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 14 (DIQMTQSPSSLSASVGDRVTITCRASGNIHNFLAWYQQKPGKAPKLLIYNEKTLADGVPSRFSGSGSGTDYTLTISSLQPEDFATYFCQQFWSTPYTFGGGTKVEIK);
a constant region of a human IgG2 isotype; and an Fc region of the constant region of the human IgG2 isotype comprising the mutations V234A and G237A,
The pharmaceutical composition is characterized in that it is used so that the anti-fractalkine antibody is administered intravenously to a human at 10 mg/kg human body weight or 15 mg/kg human body weight per administration.

In a preferred embodiment, the present invention provides a pharmaceutical composition for treating Crohn's disease, the pharmaceutical composition comprising an anti-fractalkine antibody and a pharma- ceutical acceptable excipient;
The fractalkine antibody comprises:
The amino acid sequence shown in SEQ ID NO: 11 (QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYYIHWVKQAPGQGLEWIGWIYPGDGSPKFNERFKGRTTLTADKSTNTAYMLLSSLRSEDTAVYFCATGPTDGDYFDYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVE a heavy chain consisting of: RKCCVECPPCPAPPAAAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK;
an antibody having a light chain consisting of the amino acid sequence shown in SEQ ID NO: 12 (DIQMTQSPSSLSASVGDRVTITCRASGNIHNFLAWYQQKPGKAPKLLIYNEKTLADGVPSRFSGSGSGTDYTLTISSLQPEDFATYFCQQFWSTPYTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC);
The pharmaceutical composition is characterized in that it is used so that the anti-fractalkine antibody is administered intravenously to a human in a single dose of at least 10 mg/kg of human body weight.

In another preferred embodiment, the present invention provides a pharmaceutical composition for treating Crohn's disease, the pharmaceutical composition comprising an anti-fractalkine antibody and a pharma- ceutical acceptable excipient;
The fractalkine antibody comprises:
The amino acid sequence shown in SEQ ID NO: 11 (QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYYIHWVKQAPGQGLEWIGWIYPGDGSPKFNERFKGRTTLTADKSTNTAYMLLSSLRSEDTAVYFCATGPTDGDYFDYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVE a heavy chain consisting of: RKCCVECPPCPAPPAAAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK;
an antibody having a light chain consisting of the amino acid sequence shown in SEQ ID NO: 12 (DIQMTQSPSSLSASVGDRVTITCRASGNIHNFLAWYQQKPGKAPKLLIYNEKTLADGVPSRFSGSGSGTDYTLTISSLQPEDFATYFCQQFWSTPYTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC);
The pharmaceutical composition is characterized in that it is used so that the anti-fractalkine antibody is administered intravenously to a human at 10 to 15 mg/kg of human body weight per administration.

In yet another preferred embodiment, the present invention provides a pharmaceutical composition for treating Crohn's disease, comprising an anti-fractalkine antibody and a pharma- ceutical acceptable excipient;
The fractalkine antibody comprises:
The amino acid sequence shown in SEQ ID NO: 11 (QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYYIHWVKQAPGQGLEWIGWIYPGDGSPKFNERFKGRTTLTADKSTNTAYMLLSSLRSEDTAVYFCATGPTDGDYFDYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVE a heavy chain consisting of: RKCCVECPPCPAPPAAAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK;
an antibody having a light chain consisting of the amino acid sequence shown in SEQ ID NO: 12 (DIQMTQSPSSLSASVGDRVTITCRASGNIHNFLAWYQQKPGKAPKLLIYNEKTLADGVPSRFSGSGSGTDYTLTISSLQPEDFATYFCQQFWSTPYTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC);
The pharmaceutical composition is characterized in that it is used so that the anti-fractalkine antibody is administered intravenously to a human at 10 mg/kg human body weight or 15 mg/kg human body weight per administration.

In another aspect, the pharmaceutical composition of the present invention is repeatedly administered intravenously to a human at a dosing interval of once per week to once per two weeks to provide a mean trough concentration of the anti-FKN antibody of 40 μg/mL or more. In a preferred embodiment, the pharmaceutical composition of the present invention is repeatedly administered intravenously to a human at a dosing interval of once per week to once per two weeks to provide a mean trough concentration of the anti-FKN antibody of 60 μg/mL or more. In a further preferred embodiment, the pharmaceutical composition of the present invention is repeatedly administered intravenously to a human at a dosing interval of once per week to once per two weeks to provide a mean trough concentration of the anti-FKN antibody of 80 μg/mL or more.

In a preferred embodiment, the pharmaceutical composition has a mean C _max of the anti-fractalkine antibody per 1 mg of the anti-fractalkine antibody/kg of human body weight of 21 to 25 μg/mL when administered intravenously in a single dose to humans.

In a preferred embodiment, the pharmaceutical composition is an intravenous pharmaceutical composition formulated to provide any one, two, or three of the following pharmacokinetic parameters upon a single intravenous administration of 10 mg/kg human body weight of the anti-fractalkine antibody to a human:
The mean C _max is within the range of 80% to 125% of 2.4×10 ² μg/mL;
The mean AUC _(0-t) is within the range of 80% to 125% of 7.0×10 ⁴ μg·h/mL; and
The mean AUC _{(0-336 h)} is a value included in the numerical range of 80% to 125% of 3.8×10 ⁴ μg·h/mL.

In a preferred embodiment, the pharmaceutical composition is administered intravenously to a human multiple times at intervals ranging from once per week to once every two weeks.

In a preferred embodiment, the pharmaceutical composition is administered intravenously so that the mean trough concentration of the anti-fractalkine antibody is 80 μg/mL or more.

In a preferred embodiment, the pharmaceutical composition is a pharmaceutical composition in which the anti-fractalkine antibody is administered intravenously to a human at 10 mg/kg of human body weight per administration, at least seven times at biweekly administration intervals after three weekly administrations.

In another aspect, the present invention provides a pharmaceutical composition for treating Crohn's disease, comprising:
The pharmaceutical composition comprises a therapeutically effective amount of an anti-fractalkine antibody and a pharma- ceutical acceptable excipient;
The anti-fractalkine antibody is
the heavy chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 13 (QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYYIHWVKQAPGQGLEWIGWIYPGDGSPKFNERFKGRTTLTADKSTNTAYMLLSSLRSEDTAVYFCATGPTDGDYFDYWGQGTTVTVSS);
the light chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 14 (DIQMTQSPSSLSASVGDRVTITCRASGNIHNFLAWYQQKPGKAPKLLIYNEKTLADGVPSRFSGSGSGTDYTLTISSLQPEDFATYFCQQFWSTPYTFGGGTKVEIK);
a constant region of a human IgG2 isotype; and an Fc region of the constant region of the human IgG2 isotype comprising the mutations V234A and G237A,
The therapeutically effective amount is such that, when a pharmaceutical composition containing the anti-fractalkine antibody is administered intravenously in a single dose, the mean _Cmax of the anti-fractalkine antibody per 1 mg of the anti-fractalkine antibody/kg of human body weight is 21 to 25 μg/mL.
It is a pharmaceutical composition.

In yet another aspect, the present invention provides a pharmaceutical composition for treating Crohn's disease, comprising:
The pharmaceutical composition comprises a therapeutically effective amount of an anti-fractalkine antibody and a pharma- ceutical acceptable excipient;
The anti-fractalkine antibody is
the heavy chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 13 (QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYYIHWVKQAPGQGLEWIGWIYPGDGSPKFNERFKGRTTLTADKSTNTAYMLLSSLRSEDTAVYFCATGPTDGDYFDYWGQGTTVTVSS);
the light chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 14 (DIQMTQSPSSLSASVGDRVTITCRASGNIHNFLAWYQQKPGKAPKLLIYNEKTLADGVPSRFSGSGSGTDYTLTISSLQPEDFATYFCQQFWSTPYTFGGGTKVEIK);
a constant region of a human IgG2 isotype; and an Fc region of the constant region of the human IgG2 isotype comprising the mutations V234A and G237A,
The therapeutically effective amount is determined to exhibit the following pharmacokinetic parameters when administered intravenously in a single dose of a pharmaceutical composition comprising the anti-fractalkine antibody:
The mean C _max is within the range of 80% to 125% of 2.4×10 ² μg/mL;
The mean AUC _(0-t) is within the range of 80% to 125% of 7.0×10 ⁴ μg·h/mL; and
The mean AUC _(0-336h) is within the range of 80% to 125% of 3.8 x 10 ⁴ μg·h/mL.
or
It is a pharmaceutical composition.

In a preferred embodiment, the dosage is 10-15 mg/kg human body weight of the anti-fractalkine antibody per dose, or 10 mg/kg human body weight or 15 mg/kg human body weight of the anti-fractalkine antibody per dose.

In a preferred embodiment, the pharmaceutical composition is administered intravenously to a human multiple times at intervals ranging from once per week to once every two weeks.

In a preferred embodiment, the pharmaceutical composition is administered intravenously so that the mean trough concentration of the anti-fractalkine antibody is 80 μg/mL or more.

In a preferred embodiment, the pharmaceutical composition is a pharmaceutical composition in which the anti-fractalkine antibody is administered intravenously to a human at 10 mg/kg of human body weight per administration, at least seven times at biweekly administration intervals after three weekly administrations.

In a preferred embodiment, the anti-fractalkine antibody has the amino acid sequence shown in SEQ ID NO: 11 (QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYYIHWVKQAPGQGLEWIGWIYPGDGSPKFNERFKGRTTLTADKSTNTAYMLLSSLRSEDTAVYFCATGPTDGDYFDYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPAAAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTI The antibody H3-2L4 is composed of a heavy chain consisting of SKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK) and a light chain consisting of the amino acid sequence shown in SEQ ID NO: 12 (DIQMTQSPSSLSASVGDRVTITCRASGNIHNFLAWYQQKPGKAPKLLIYNEKTLADGVPSRFSGSGSGTDYTLTISSLQPEDFATYFCQQFWSTPYTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC).

In another aspect, the present invention provides a therapeutic agent for Crohn's disease comprising an anti-fractalkine antibody,
The method is characterized in that an anti-fractalkine antibody is intravenously administered to a human at a dose of 10 to 15 mg/kg of human body weight per administration at an administration interval of once a week to once every two weeks;
The anti-fractalkine antibody is
the heavy chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 13 (QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYYIHWVKQAPGQGLEWIGWIYPGDGSPKFNERFKGRTTLTADKSTNTAYMLLSSLRSEDTAVYFCATGPTDGDYFDYWGQGTTVTVSS);
the light chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 14 (DIQMTQSPSSLSASVGDRVTITCRASGNIHNFLAWYQQKPGKAPKLLIYNEKTLADGVPSRFSGSGSGTDYTLTISSLQPEDFATYFCQQFWSTPYTFGGGTKVEIK);
a constant region of a human IgG2 isotype; and an Fc region of the constant region of the human IgG2 isotype comprising the mutations V234A and G237A.
It is a drug used to treat Crohn's disease.

In one embodiment of the present invention, there is provided a therapeutic agent for Crohn's disease comprising an anti-fractalkine antibody,
The method is characterized in that an anti-fractalkine antibody is intravenously administered to a human at a dose of 10 mg/kg of human body weight or 15 mg/kg of human body weight at an administration interval of once a week to once every two weeks;
The anti-fractalkine antibody is
the heavy chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 13 (QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYYIHWVKQAPGQGLEWIGWIYPGDGSPKFNERFKGRTTLTADKSTNTAYMLLSSLRSEDTAVYFCATGPTDGDYFDYWGQGTTVTVSS);
the light chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 14 (DIQMTQSPSSLSASVGDRVTITCRASGNIHNFLAWYQQKPGKAPKLLIYNEKTLADGVPSRFSGSGSGTDYTLTISSLQPEDFATYFCQQFWSTPYTFGGGTKVEIK);
a constant region of a human IgG2 isotype; and an Fc region of the constant region of the human IgG2 isotype comprising the mutations V234A and G237A.
It is a pharmaceutical composition.

In another preferred embodiment, the present invention provides a therapeutic agent for Crohn's disease comprising an anti-fractalkine antibody,
The method is characterized in that an anti-fractalkine antibody is intravenously administered to a human at a dose of 10 to 15 mg/kg of human body weight per administration at an administration interval of once a week to once every two weeks;
The anti-fractalkine antibody is
The amino acid sequence shown in SEQ ID NO: 11 (QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYYIHWVKQAPGQGLEWIGWIYPGDGSPKFNERFKGRTTLTADKSTNTAYMLLSSLRSEDTAVYFCATGPTDGDYFDYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVE a heavy chain consisting of: RKCCVECPPCPAPPAAAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK;
an antibody having a light chain consisting of the amino acid sequence shown in SEQ ID NO: 12 (DIQMTQSPSSLSASVGDRVTITCRASGNIHNFLAWYQQKPGKAPKLLIYNEKTLADGVPSRFSGSGSGTDYTLTISSLQPEDFATYFCQQFWSTPYTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC);
It is a drug used to treat Crohn's disease.

In one embodiment of the present invention, there is provided a therapeutic agent for Crohn's disease comprising an anti-fractalkine antibody,
The method is characterized in that an anti-fractalkine antibody is intravenously administered to a human at a dose of 10 mg/kg of human body weight or 15 mg/kg of human body weight at an administration interval of once a week to once every two weeks;
The anti-fractalkine antibody is
The amino acid sequence shown in SEQ ID NO: 11 (QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYYIHWVKQAPGQGLEWIGWIYPGDGSPKFNERFKGRTTLTADKSTNTAYMLLSSLRSEDTAVYFCATGPTDGDYFDYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVE a heavy chain consisting of: RKCCVECPPCPAPPAAAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK;
an antibody having a light chain consisting of the amino acid sequence shown in SEQ ID NO: 12 (DIQMTQSPSSLSASVGDRVTITCRASGNIHNFLAWYQQKPGKAPKLLIYNEKTLADGVPSRFSGSGSGTDYTLTISSLQPEDFATYFCQQFWSTPYTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC);
It is a drug used to treat Crohn's disease.

In one embodiment of the present invention, the pharmaceutical composition of the present invention is a pharmaceutical composition in which, when the anti-fractalkine antibody is administered intravenously to a human at a dose of at least 10 mg/kg of human body weight multiple times at intervals of once per week to once per two weeks, the average serum total FKN concentration is 200 ng/mL or more, 300 ng/mL or more, or 400 ng/mL or more.

In another embodiment of the present invention, the pharmaceutical composition of the present invention is a pharmaceutical composition in which, when the anti-fractalkine antibody is administered intravenously to a human at a dose of 10 to 15 mg/kg of human body weight multiple times at intervals of once a week to once every two weeks, the average serum total FKN concentration is 200 ng/mL or more, 300 ng/mL or more, or 400 ng/mL or more.

In another embodiment of the present invention, the pharmaceutical composition of the present invention is a pharmaceutical composition in which, when the anti-fractalkine antibody is administered intravenously to a human at a dose of 10 mg/kg human body weight or 15 mg/kg human body weight multiple times at administration intervals of once a week to once every two weeks, the mean serum total FKN concentration is 200 ng/mL or more, 300 ng/mL or more, or 400 ng/mL or more.

In one embodiment of the present invention, the pharmaceutical composition of the present invention is a pharmaceutical composition that, when the anti-fractalkine antibody is administered intravenously to a human at a dose of at least 10 mg/kg of human body weight multiple times at intervals of once a week to once every two weeks, reduces the CDAI by 70 or more, preferably 100 or more, from the score before the start of administration 12 weeks or later.

In another embodiment of the present invention, the pharmaceutical composition of the present invention is a pharmaceutical composition that, when the anti-fractalkine antibody is administered intravenously to a human at 10 to 15 mg/kg of human body weight per administration multiple times at intervals ranging from once a week to once every two weeks, reduces the CDAI by 70 or more, preferably 100 or more, from the score before the start of administration 12 weeks or later.

In another embodiment of the present invention, the pharmaceutical composition of the present invention is a pharmaceutical composition that, when the anti-fractalkine antibody is administered intravenously to a human at 10 mg/kg human body weight or 15 mg/kg human body weight multiple times at intervals of once a week to once every two weeks, reduces the CDAI by 70 or more, preferably 100 or more, from the score before the start of administration 12 weeks or later.

In a preferred embodiment, the pharmaceutical composition of the present invention is a pharmaceutical composition that, when the anti-fractalkine antibody is administered intravenously multiple times at an administration interval of at least 10 mg/kg of human body weight per administration to a human with a CDAI score of 220 or more before the start of administration, from once a week to once every two weeks, reduces the CDAI by 70 or more, preferably 100 or more, from the score at the start of administration 12 weeks or later.

In another preferred embodiment, the pharmaceutical composition of the present invention is a pharmaceutical composition in which, when the anti-fractalkine antibody is administered intravenously multiple times at intervals of once a week to once every two weeks at 10 to 15 mg/kg of human body weight per administration to a human with a CDAI score of 220 or more before the start of administration, the CDAI score is reduced by 70 or more, preferably 100 or more, from the score at the start of administration 12 weeks or later.

In yet another preferred embodiment, the pharmaceutical composition of the present invention is a pharmaceutical composition in which, when the anti-fractalkine antibody is administered intravenously multiple times at intervals of once a week to once every two weeks at 10 mg/kg human body weight or 15 mg/kg human body weight per administration to a human with a CDAI score of 220 or more before the start of administration, the CDAI score is reduced by 70 or more, preferably 100 or more, from the score at the start of administration 12 weeks or later.

In a preferred embodiment, the pharmaceutical composition of the present invention is a pharmaceutical composition that, when the anti-fractalkine antibody is administered intravenously multiple times at an administration interval of at least 10 mg/kg of human body weight per administration to a human with a CDAI score of 220 or more before the start of administration, the CDAI is less than 150 12 weeks or more after the start of administration.

In another preferred embodiment, the pharmaceutical composition of the present invention is a pharmaceutical composition in which, when the anti-fractalkine antibody is administered intravenously multiple times at intervals of once a week to once every two weeks at 10 to 15 mg/kg of human body weight per administration to a human with a CDAI score of 220 or more before the start of administration, the CDAI is less than 150 12 weeks or more after the start of administration.

In yet another preferred embodiment, the pharmaceutical composition of the present invention is a pharmaceutical composition in which, when the anti-fractalkine antibody is administered intravenously multiple times at intervals of once a week to once every two weeks to a human with a CDAI score of 220 or more before the start of administration at 10 mg/kg human body weight or 15 mg/kg human body weight per administration, the CDAI is less than 150 12 weeks or more after the start of administration.

In one embodiment of the invention, the "human" is a patient with Crohn's disease who has had an inadequate, non-sustained, or intolerant response to corticosteroids or immunomodulatory drugs, or who has had an inadequate response to anti-TNF antibodies.

A person skilled in the art will understand that the present invention may be implemented by appropriately combining any one or more of the aspects described in this specification, unless technically inconsistent. Furthermore, a person skilled in the art will understand that it would be preferable to implement the present invention by appropriately combining any preferred or advantageous aspects described in this specification, unless technically inconsistent.

The documents cited in this specification should be considered to be expressly incorporated herein in their entirety by reference, and a person skilled in the art would be able to understand, in accordance with the context of this specification, the relevant disclosures in those documents as incorporated as part of this specification without departing from the spirit and scope of the present invention.

The documents cited herein are provided solely for the purpose of disclosing relevant technology prior to the filing date of this application and should not be construed as an admission that the inventors are not entitled to antedate such disclosure by virtue of prior invention or for any other reason. All statements in these documents are based on information available to the applicant and do not constitute any admission that the contents of such statements are accurate.

The terms used in this specification are used to describe specific embodiments and are not intended to limit the invention.

The term "comprise" or "include" as used herein intends that the stated items (such as components, steps, elements, or numbers) are present, unless the context clearly dictates otherwise, and does not exclude the presence of other items (such as components, steps, elements, or numbers). The term "consist of" encompasses aspects described with the terms "consist of" and/or "consist essentially of".

Unless otherwise defined, all terms used herein (including technical and scientific terms) have the same meaning as commonly understood by those skilled in the art to which this invention belongs. Terms used herein should be interpreted as having a meaning consistent with the meaning in this specification and related technical fields, and should not be interpreted in an idealized or overly formal sense, unless otherwise defined.

Although terms such as first, second, etc. are used to describe various elements, it is understood that these elements should not be limited by these terms themselves. These terms are used only to distinguish one element from another element, and for example, it is possible to refer to a first element as a second element and similarly, to refer to a second element as a first element without departing from the scope of the present invention.

In this specification, numerical values used to indicate ingredient contents, numerical ranges, etc. should be understood to be modified with the term "about" unless otherwise specified. For example, "10 μg" is understood to mean "about 10 μg" unless otherwise specified, and it is natural that a person skilled in the art would be able to reasonably understand that level in accordance with common technical knowledge and the meaning of this specification.

Unless the context clearly indicates otherwise, as used in this specification and claims, each aspect expressed in the singular may also be expressed in the plural, and vice versa, unless technically inconsistent.

The present invention will be described in more detail below with reference to examples. However, the present invention can be embodied in various ways and should not be construed as being limited to the examples described herein. Those skilled in the relevant art can implement the present invention with various modifications, additions, deletions, substitutions, etc. without altering the spirit or scope of the present invention.

The abbreviations used in the examples are conventional abbreviations well known to those skilled in the art. Some abbreviations are listed below.
AUC: area under the serum concentration-time curve
AUC _(0-inf) : Area under the serum concentration-time curve from zero time extrapolated to infinite time
AUC _(0-t) : Area under the concentration-time curve from zero time to time of last quantifiable concentration
AUC _(0-336h) : Area under the concentration-time curve from zero (predose) to fixed time-point 336 h (2 weeks) after the end of infusion
CDAI: Crohn's disease activity index
CL: total clearance
CRP: C-reactive protein
_Cmax : maximum serum concentration
ELISA: enzyme-linked immunosorbent assay
FKN: fractalkine
IgG: immunoglobulin G
NONMEN: nonlinear mixed effect model
SAS: statistical analysis system
t _1/2 : serum elimination half-life
t _max : time to reach peak serum concentration
_Vd : volume of distribution

Example 1: Preparation of humanized anti-human fractalkine antibody In the following human administration test, humanized anti-human fractalkine antibody H3-2L4 was used. H3-2L4, including humanization, was produced as described in WO2011/052799. H3-2L4 used in Example 2 and onwards was prepared by the methods described in 1-1 and 1-2 below.

1-1. Expression Vector An expression vector for the humanized anti-human fractalkine antibody (H3-2L4) was constructed as follows.
First, a signal sequence (SEQ ID NO: 3) was added to the N-terminus of the amino acid sequence (SEQ ID NO: 1) of the heavy chain variable region (H3-2) of the humanized anti-human fractalkine antibody (H3-2L4), and the amino acid sequence (SEQ ID NO: 4) of the constant region of human IgG2 into which two mutations (V234A and G237A) were inserted was added to the C-terminus (SEQ ID NO: 5). Next, a signal sequence (SEQ ID NO: 6) was added to the N-terminus of the amino acid sequence (SEQ ID NO: 2) of the light chain variable region (L4) of the humanized anti-human fractalkine antibody (H3-2L4), and the amino acid sequence (SEQ ID NO: 7) of the constant region of human Igκ was added to the C-terminus (SEQ ID NO: 8). These amino acid sequences (SEQ ID NOs: 5 and 8) were converted into optimal gene sequences for expression in CHO cells, and sequences were totally synthesized by adding the recognition sequence for the restriction enzyme HindIII and the Kozak sequence to the 5'-terminus of the gene sequence, and a stop codon and the recognition sequence for the restriction enzyme EcoRI to the 3'-terminus (SEQ ID NOs: 9 and 10).
The sequences identified by SEQ ID NOs: 1 to 10 are as follows:

Amino acid sequence of heavy chain variable region (H3-2) (SEQ ID NO:1)
QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYYIHWVKQAPGQGLEWIGWIYPGDGSPKFNERFKGRTTLTADKSTNTAYMLLSSLRSEDTAVYFCATGPTDGDYFDYWGQGTTVTVSS

Amino acid sequence of the light chain variable region (L4) (SEQ ID NO:2)
DIQMTQSPSSLSASVGDRVTITCRASGNIHNFLAWYQQKPGKAPKLLIYNEKTLADGVPSRFSGSGSGTDYTLTISSLQPEDFATYFCQQFWSTPYTFGGGTKVEIK

Signal sequence (SEQ ID NO:3)
MEWSWVFLFFLSVTTGVHS

Amino acid sequence of the constant region of human IgG2 with two mutations (V234A and G237A) (SEQ ID NO:4)
ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPAAAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

(SEQ ID NO:5)
MEWSWVFLFFLSVTTGVHSQVQLVQSGAEVKKPGASVKVSCKASGYTFTNYYIHWVKQAPGQGLEWIGWIYPGDGSPKFNERFKGRTTLTADKSTNTAYMLLSSLRSEDTAVYFCATGPTDGDYFDYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKV DKTVERKCCVECPPCPAPPAAAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

Signal sequence (SEQ ID NO:6)
MSVPTQVLGLLLLWLTDARC

Amino acid sequence of the constant region of human Igκ (SEQ ID NO:7)
RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

(SEQ ID NO:8)
MSVPTQVLGLLLLWLTDARCDIQMTQSPSSLSASVGDRVTITCRASGNIHNFLAWYQQKPGKAPKLLIYNEKTLADGVPSRFSGSGSGTDYTLTISSLQPEDFATYFCQQFWSTPYTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

(SEQ ID NO:9)

(SEQ ID NO: 10)

The gene sequence encoding the fully synthesized heavy chain was excised with the restriction enzymes HindIII and EcoRI and inserted into the HindIII and EcoRI sites of the pEE6.4 vector (Lonza). The gene sequence encoding the fully synthesized light chain was excised with the restriction enzymes HindIII and EcoRI and inserted into the HindIII and EcoRI sites of the pEE12.4 vector (Lonza). Each vector was cleaved with the restriction enzymes NotI and PvuI, and the vector fragments containing the heavy chain and light chain were ligated together to construct an expression vector.

1-2. Construction of a cell line expressing antibody H3-2L4 and acquisition of antibody H3-2L4 The prepared expression vector was introduced by electroporation into CHOK1SV cells (Lonza) acclimatized to CD-CHO/6 mM L-glutamine medium. After gene introduction, selection was performed in CD-CHO/50 uM MSX medium at 37°C/10% _CO2 to obtain cells expressing the target antibody. Then, cell cloning was performed to prepare a cell bank. The prepared cell bank was resuscitated and cultured, and the culture supernatant was purified by chromatography to obtain the target antibody H3-2L4.

The antibody H3-2L4 obtained as described above had heavy and light chains with the amino acid sequences shown in SEQ ID NOs: 11 and 12, respectively.

Full length heavy chain of H3-2L4 (SEQ ID NO:11)
QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYYIHWVKQAPGQGLEWIGWIYPGDGSPKFNERFKGRTTLTADKSTNTAYMLLSSLRSEDTAVYFCATGPTDGDYFDYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCC VECPPCPAPPAAAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

Full length light chain of H3-2L4 (SEQ ID NO: 12)
DIQMTQSPSSLSASVGDRVTITCRASGNIHNFLAWYQQKPGKAPKLLIYNEKTLADGVPSRFSGSGSGTDYTLTISSLQPEDFATYFCQQFWSTPYTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

The amino acid sequences of the heavy chain variable region and light chain variable region of antibody H3-2L4 were as follows:

Heavy chain variable region of H3-2L4 (SEQ ID NO: 13)
QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYYIHWVKQAPGQGLEWIGWIYPGDGSPKFNERFKGRTTLTADKSTNTAYMLLSSLRSEDTAVYFCATGPTDGDYFDYWGQGTTVTVSS

Light chain variable region of H3-2L4 (SEQ ID NO: 14)
DIQMTQSPSSLSASVGDRVTITCRASGNIHNFLAWYQQKPGKAPKLLIYNEKTLADGVPSRFSGSGSGTDYTLTISSLQPEDFATYFCQQFWSTPYTFGGGTKVEIK

The amino acid sequences of CDR-H1 to CDR-H3 and CDR-L1 to CDR-L3 of antibody H3-2L4 were as follows:

CDR-H1 of H3-2L4 (SEQ ID NO: 15)
NYYIH

CDR-H2 of H3-2L4 (SEQ ID NO: 16)
WIYPGDGSPKFNERFKG

CDR-H3 of H3-2L4 (SEQ ID NO: 17)
GPTDGDYFDY

CDR-L1 of H3-2L4 (SEQ ID NO: 18)
RASGNIHNFLA

CDR-L2 of H3-2L4 (SEQ ID NO: 19)
NEKTLAD

CDR-L3 of H3-2L4 (SEQ ID NO:20)
QQFWSTPYT

Example 2: Single Dose Study A Phase 1 clinical study was conducted to evaluate the safety and tolerability, as well as the pharmacokinetics, of a single intravenous dose of H3-2L4 in healthy adult male subjects.

2-1. [Clinical trial design]
This study was a single-center, randomized, double-blind, placebo-controlled, single-dose ascending study with the primary objective of evaluating the safety and tolerability of a single intravenous dose of H3-2L4 in healthy adult male Japanese subjects. Sixty-four subjects were divided into eight cohorts (0.0006, 0.006, 0.04, 0.2, 1, 3, 6, and 10 mg/kg groups), and six of the eight subjects in each cohort received a single intravenous dose of H3-2L4 and two received a placebo.

This clinical trial consisted of a screening period, an observational inpatient period, an inpatient administration period (double-blind), follow-up period 1 (double-blind), and follow-up period 2 (only cohorts 5 to 8: open-label).

Screening tests were performed within 2 to 27 days prior to administration of the study drug, and tests during the observational hospitalization period were performed the day before administration to confirm eligibility. Subjects who were confirmed as eligible were randomly assigned to the H3-2L4 or placebo group based on an allocation table prepared by the person in charge of allocation. Two subjects in each cohort were assigned to receive H3-2L4 and one placebo on the first day. The remaining six subjects were assigned to receive H3-2L4 and one placebo on the second to fourth days. The number of subjects administered on the same day was limited to two, and the second was administered at least two hours after the start of administration of the first. The study drug was administered by intravenous drip infusion over approximately 30 minutes after dissolving H3-2L4 or placebo in saline solution. This was a single-dose study, and subjects were administered only once, regardless of meal times.

After administration, subjects were observed and investigated as hospitalized patients for one week, and then observed and investigated as outpatients until 8 weeks after the end of administration in cohorts 1 to 4 (0.0006, 0.006, 0.04), and until 24 weeks after the end of administration in cohorts 5 to 8 (0.2, 1, 3, 6, 10 mg/kg groups).

Cohort transitions were made when the principal investigator and sponsor (medical experts, the sponsor's safety department staff, and the sponsor's representative) both determined that there were no problems with the transition based on the safety results obtained in each cohort, and were generally carried out every three weeks from the administration of the first subject in the previous cohort.

The types of investigational drugs used in this study were as follows:

2-2. [Administration of investigational drug]
H3-2L4 or placebo was diluted with physiological saline according to the body weight of each subject to prepare the administration volume shown in Table 2. For administration, a syringe pump or infusion pump was used to administer the drug intravenously using an in-line filter (pore size: 0.2 μm) for approximately 30 minutes (including saline flush).

2-3. [Analysis of serum H3-2L4 concentration]
The serum H3-2L4 concentration was measured using a validated measurement method. Specifically, serum H3-2L4 was bound to human fractalkine immobilized on a microplate, reacted with ruthenium-labeled anti-H3-2L4 rabbit polyclonal antibody, and then quantified by measuring the amount of electrochemiluminescence using a Sector Imager 6000 (Meso Scale Discovery). Blood samples for measuring serum H3-2L4 concentration were taken at the blood sampling times shown in Table 3 below.

2-4. [Pharmacokinetic analysis]
Pharmacokinetic analysis was performed using a group of subjects who had data that allowed calculation of one or more pharmacokinetic parameters based on serum H3-2L4 concentration data (hereinafter referred to as the pharmacokinetic analysis population). In this clinical trial, 16 subjects who received a placebo and 6 subjects who received 0.0006 mg/kg and 0.006 mg/kg H3-2L4 had serum H3-2L4 concentrations below the lower limit of quantification (0.100 μg/mL), so the remaining 36 subjects were excluded from the pharmacokinetic analysis population. Summary statistics of serum H3-2L4 concentrations were calculated for each dose at each specified time. A serum H3-2L4 concentration time course graph was prepared, and pharmacokinetic parameters including C _max , t _max , AUC _(0-t) , AUC _(0-inf) , t _1/2 , CL, and V _d were calculated by noncompartmental analysis using the serum H3-2L4 concentration. In addition, dose proportionality was examined using the obtained C _max , AUC _(0-t) , and AUC _(0-inf) . The analysis was performed using SAS, WinNonlin, Pharsight Knowledgebase Server, Microsoft Excel, and S-PLUS.

The results calculated as above are shown in Table 4 below and Figure 1. The average values of the pharmacokinetic parameters other than t _max in the table are shown as arithmetic means.

As a result of the analysis, the pharmacokinetics of H3-2L4 when administered intravenously once was considered to show a triphasic profile of α-phase, β-phase, and γ-phase at 0.04-10 mg/kg. After the end of intravenous infusion, an initial distribution phase (α-phase) in which the concentration rapidly decreased from _Cmax was observed, followed by a gradual elimination phase (β-phase) at high concentrations and a steep final elimination phase (γ-phase) at low concentrations. The boundary between the γ-phase and β-phase was considered to be around 10 μg/mL. In the 0.04 and 0.2 mg/kg groups, a biphasic profile was observed, and the α-phase was followed by the γ-phase. In the 1-10 mg/kg group, the number of blood samples required to accurately evaluate the γ-phase, which is the final elimination phase, was insufficient, so pharmacokinetic parameters dependent on the γ-phase could not be calculated. After 30 minutes of intravenous infusion, most subjects reached _tmax 0.5 hours later, but some subjects reached _tmax 6.5 hours later. t _1/2 , _β was longer than t _1/2 , _γ and increased with increasing dose, whereas t _1/2 , _β in the 6 and 10 mg/kg groups were similar and appeared to have reached a plateau. C _max increased proportionally to dose from 0.04 to 10 mg/kg, whereas AUC _(0-t) and AUC _(0-inf) were not proportional to dose, indicating that H3-2L4 exhibited nonlinear pharmacokinetics.

Example 3: Multiple-dose study A Phase 1/2 clinical study was conducted to evaluate the safety, tolerability, pharmacokinetics, and efficacy against Crohn's disease when H3-2L4 was administered intravenously multiple times in patients with Crohn's disease.

3-1. [Clinical trial design]
This clinical trial was a multicenter, open-label, non-controlled, multiple ascending dose (MAD) study with the primary objective of evaluating the safety and tolerability of repeated intravenous administration of H3-2L4 in Japanese patients with Crohn's disease, and was not randomized. In this clinical trial, 28 subjects were divided into four cohorts 1 to 4 (2, 5, 10, and 15 mg/kg groups) of 6, 8, 7, and 7 subjects, respectively, and received repeated intravenous administration of H3-2L4.
This clinical trial consisted of a screening period, an observation period, an administration period, a continuation administration period, and a follow-up period.

Screening tests were conducted within 2 to 42 days prior to the start of administration of the study drug, and observational tests were conducted the day before or on the day of the first administration of the study drug, and H3-2L4 was administered to subjects whose eligibility was confirmed.

Eligibility selection criteria were as follows:
(1) Be at least 20 years old and under 65 years old;
(2) Patients who have been diagnosed with Crohn's disease according to the Crohn's disease diagnostic criteria (2012) established by the "Study on Intractable Inflammatory Bowel Disorders" group, part of the Research Project to Overcome Intractable Diseases funded by the Ministry of Health, Labor and Welfare's Science Research Grant.
(3) Patients with mild to moderate disease severity during the observation period (CDAI score of 150 or greater but less than 450, according to the same Crohn's disease diagnostic criteria).
(4) Patients who have previously been administered aminosalicylic acid preparations (5-ASA), salazosulfapyridine, corticosteroids, immunomodulators, infliximab (including biosimilars, the same applies below), or adalimumab and have not seen any effect, or whose effect was weakened or disappeared after being administered, or who have been unable to continue administration due to side effects (excluding infliximab and adalimumab).

H3-2L4 was dissolved in approximately 100 mL of physiological saline solution and administered intravenously over approximately 30 minutes. During the administration period, H3-2L4 was administered every 2 weeks for the first (2 mg/kg) and second cohorts (5 mg/kg) for a total of 6 times up to week 10 (double dose at week 0), and H3-2L4 was administered at weeks 0, 1, and 2, and then every 2 weeks for a total of 7 times up to week 10 for the third (10 mg/kg) and fourth cohorts (15 mg/kg). In the evaluation at week 12, subjects who had no safety issues, a Crohn's Disease Activity Index (CDAI) score of less than 150 or a decrease of 70 or more from the observation period, and who wished to continue administration were administered the same dose 20 times every 2 weeks (40 weeks) (continuation administration period). During the continued administration period, the principal investigator or subinvestigator may decide to change (reduce) the dose to every four weeks at the same dose (it may also be possible to revert to administration every two weeks).

Cohort transitions were made when both the investigator and sponsor judged there to be no problem with the transition based on the safety results obtained in each cohort.

The types of investigational drugs used in this study were as follows:

3-2. [Administration of investigational drug]
H3-2L4 was diluted with physiological saline according to the subject's weight to prepare the administration volume (approximately 100 mL) of H3-2L4 at each dose. The doses (2, 5, 10, and 15 mg/kg) and administration methods are shown in Table 6. The body weight used to calculate the dose (total amount) converted into body weight was the body weight measured at the observation period, 12 weeks, 24 weeks, and 36 weeks, and the converted dose (total amount) was not changed even if the body weight increased or decreased during that period. When administering, an infusion pump was used to administer the drug intravenously for approximately 30 minutes (including saline flush) using an in-line filter (pore size: 0.2 μm). The investigational drug could be administered regardless of meal times.

3-3. [Pharmacokinetics and pharmacodynamics]
Using a validated measurement method, serum H3-2L4 concentration and serum total FKN concentration were measured. Serum H3-2L4 concentration was measured by the method described in Example 2, 2-4. Serum total FKN concentration was measured by sandwich ELISA using two types of anti-human fractalkine monoclonal antibodies 1F3 and 3A5-2 described in WO2011/052799. Specifically, total FKN in serum was captured with 1F3 immobilized on a plate, sandwiched with 3A5-2 labeled with horseradish peroxidase (HRP), and further reacted with 3,3',5,5'-tetramethylbenzidine (TMB), which is a substrate for HRP, and the color development value was measured with an autoreader (Microplate reader Envision, PerkinElmer Japan), thereby quantifying the serum total FKN concentration.

Pharmacokinetic analysis was performed based on serum H3-2L4 concentration data using a group of subjects who were administered the investigational drug and had at least one evaluable serum EH3-2L4 concentration data point (hereafter referred to as the pharmacokinetic analysis population). Summary statistics of serum H3-2L4 concentrations were calculated at specified times by dose, and a graph of serum H3-2L4 concentration over time was created.

Pharmacodynamic analysis was performed using a group of subjects who were administered the investigational drug and had at least one evaluable pharmacodynamic biomarker data point (hereafter referred to as the pharmacodynamic analysis group). Summary statistics were calculated for the measured serum total FKN concentrations by dose at each evaluation time point.

3-3-1. [Serum H3-2L4 concentration and serum total FKN concentration]
From the time of administration of the investigational drug until 12 weeks, blood samples were taken for measurement of serum H3-2L4 concentration and serum total FKN concentration before administration and within 10 minutes at the end of administration (0 h) at the time marked with an X in the implementation schedule in Table 7 for cohorts 1 and 2, and in Table 8 for cohorts 3 and 4. For subjects who moved to the continuous administration period, blood samples were taken every 4 weeks until the 52nd week. In addition, for subjects who discontinued the clinical trial during the administration period or the continuous administration period, blood samples were taken at the time of discontinuation.

The mean values (expressed as arithmetic means) of serum H3-2L4 concentrations and serum total FKN concentrations in the 2 mg/kg, 5 mg/kg, 10 mg/kg, and 15 mg/kg groups over the administration period (12 weeks) are shown in Tables 9 and 10, respectively. The time courses of the mean values of serum H3-2L4 concentrations and serum total FKN concentrations are shown in Figures 2 and 3, respectively.

3-4. [Binding Occupancy]
The serum H3-2L4 concentration and serum total FKN concentration in the single-dose administration test of H3-2L4 in healthy Japanese adults in Example 2 were measured using a two-target quasi-steady-state (QSS) model (J Pharmacokinet Pharmacodyn 2012;39:217-26) to estimate the binding affinity parameter (K ^m _ss ). The rate was simulated using the serum H3-2L4 concentration and _{the corresponding K mss} ^. The free FKN (cFKN) concentration in blood was calculated using the serum H3-2L4 concentration, serum total FKN concentration, and the QSS for cFKN. The simulation was performed using _a constant (K ^css ). It was assumed that the QSS constants were the same between healthy adults and Crohn's disease patients.

Figures 4 and 5 show the time-dependent changes in the binding occupancy rate for mFKN by dose and the ratio of cFKN amount to the baseline amount of cFKN before the start of administration in each administration group, based on the above simulation. These results suggest that 10 mg/kg and 15 mg/kg of anti-fractalkine antibody are effective doses per administration for both binding to mFKN and inhibiting cFKN.

3-5. [Effectiveness]
Efficacy analysis was performed using the full analysis set (FAS: Full Analysis Set), which is the set of subjects who were administered the investigational drug and had at least one evaluable efficacy data point after administration of the investigational drug.

Summary statistics for CDAI and CRP were calculated for each dose at each evaluation time point.

The analysis was carried out using WinNonlin, NONMEM, SAS, etc.

[CDAI]
CDAI was calculated from the subject's diary and the corresponding test results at the time marked with an X in the implementation schedule (Tables 7 and 8). In addition, for subjects who entered the continuous administration period, CDAI was calculated in the same manner every 4 weeks until Week 52. When calculating CDAI, the values on the day of evaluation were used for hematocrit value and weight, and the most recent value measured according to the implementation schedule was used for height.
A CDAI decrease of 70 or more from the observation period was defined as improvement, and a CDAI of less than 150 was defined as remission.
At the start of this clinical trial, there were 6, 8, 7, and 7 subjects in cohorts 1 to 4, respectively, but by 12 weeks after the first administration, 3, 2, 2, and 2 subjects in cohorts 1 to 4, respectively, discontinued the clinical trial. All subjects in cohorts 1 to 4, except for one subject in cohort 2, had a history of use of anti-TNF antibodies. Two, five, three, and two subjects, respectively, used one drug, and four, two, four, and five subjects, respectively, used two drugs, and in all cases the effect of use of anti-TNF antibodies was insufficient.

Among subjects with a CDAI of 220 or more before administration of the study drug, 1, 2, 4, and 3 subjects in cohorts 1 to 4, respectively, showed a decrease of 70 or more (CR-70) from the baseline CDAI before administration of the study drug at the 12-week evaluation. In addition, 1, 1, 4, and 3 subjects in cohorts 1 to 4, respectively, showed a decrease of 100 or more (CR-100). In addition, 3 subjects in cohort 3 and 1 subject in cohort 4 achieved remission. Of the 7 subjects in cohort 4, 3 had a CDAI of 330 or more before administration of the study drug. When stratified by subjects with a CDAI of less than 330 before administration of the study drug, 4 of 6 subjects (66%) in cohort 3 and 3 of 4 subjects (75%) in cohort 4 showed a decrease of 100 or more (CR-100). These results suggest that 10 mg/kg and 15 mg/kg of anti-fractalkine antibody are single doses that bring about good improvement effects in Crohn's disease.

Furthermore, one, two, and two subjects in cohorts 1 to 3, respectively, completed the extension treatment period. With the exception of one subject in cohort 2, all subjects who completed the extension treatment period maintained improvement throughout the 52-week period. At the 52-week evaluation, one, one, and two subjects in cohorts 1 to 3, respectively, achieved a reduction of 100 or more (CR-100) in CDAI before administration of the study drug, using the CDAI as baseline. Furthermore, two subjects in cohort 3 achieved remission.

In addition, in cohort 4, three subjects who showed improvement in the 12-week evaluation moved on to the continued treatment period, and two of these subjects maintained the improvement.

[CRP]
CRP was measured at the times marked with X in the implementation schedule (Tables 7 and 8).
The time course of CRP in seven subjects in Cohort 3 over 12 weeks is shown in Figure 6. At the 12-week evaluation, three subjects who achieved remission with CDAI (subjects 1, 3, and 6) were found to also have low CRP values.

Claims (10)

A pharmaceutical composition for treating Crohn's disease, comprising:
The pharmaceutical composition comprises an anti-fractalkine antibody and a pharma- ceutically acceptable excipient;
The anti-fractalkine antibody is
the heavy chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 13 (QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYYIHWVKQAPGQGLEWIGWIYPGDGSPKFNERFKGRTTLTADKSTNTAYMLLSSLRSEDTAVYFCATGPTDGDYFDYWGQGTTVTVSS);
the light chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 14 (DIQMTQSPSSLSASVGDRVTITCRASGNIHNFLAWYQQKPGKAPKLLIYNEKTLADGVPSRFSGSGSGTDYTLTISSLQPEDFATYFCQQFWSTPYTFGGGTKVEIK);
a constant region of a human IgG2 isotype; and an Fc region of the constant region of the human IgG2 isotype comprising the mutations V234A and G237A,
The pharmaceutical composition is characterized in that the anti-fractalkine antibody is intravenously administered to a human patient with Crohn's disease having a Crohn's disease activity index (CDAI) of 220 or more and less than 330 at a dose of 10 to 15 mg/kg of human body weight per administration so that a mean trough concentration is 80 μg/mL or more.
The pharmaceutical composition. 2. The pharmaceutical composition of claim 1,
The pharmaceutical composition, wherein the Crohn's disease is Crohn's disease for which a sufficient therapeutic effect has not been obtained, the therapeutic effect has not been sustained, or the patient has been intolerant to at least one standard treatment with a drug selected from a corticosteroid, an immunomodulatory agent, and/or an anti-TNF antibody. 2. The pharmaceutical composition of claim 1,
The patient has Crohn's disease in which aminosalicylic acid preparations, salazosulfapyridine, corticosteroids, immunomodulators, infliximab or adalimumab have been administered in the past without any effect, or the effect was weakened or disappeared after it was administered, or the administration could not be continued due to side effects.
The pharmaceutical composition. The pharmaceutical composition according to any one of claims 1 to 3,
The pharmaceutical composition is used so that the anti-fractalkine antibody is intravenously administered to the human patient at a dose of 10 mg/kg human body weight or 15 mg/kg human body weight per administration.
The pharmaceutical composition. The pharmaceutical composition according to any one of claims 1 to 3,
the mean Cmax of the anti-fractalkine antibody per mg of the anti-fractalkine antibody/kg of human body weight is 21 to 25 μg/mL when administered intravenously in a single dose to the human patient;
The pharmaceutical composition. The pharmaceutical composition according to any one of claims 1 to 3,
the mean Cmax of the anti-fractalkine antibody is within a numerical range of 80% to 125% of 2.4×10 ² μg/mL when the anti-fractalkine antibody is administered intravenously in a single dose of 10 mg/kg of human body weight to the human patient;
The pharmaceutical composition. The pharmaceutical composition according to any one of claims 1 to 3,
When the anti-fractalkine antibody is administered intravenously once at 10 mg/kg of human body weight to the human patient, the mean AUC(0-t) of the anti-fractalkine antibody is a value falling within a numerical range of 80% to 125% of 7.0×10 ⁴ μg·h/mL.
The pharmaceutical composition. The pharmaceutical composition according to any one of claims 1 to 3,
When the anti-fractalkine antibody is administered intravenously in a single dose of 10 mg/kg of human body weight to the human patient, the mean AUC (0-336 h) of the anti-fractalkine antibody is a value falling within a numerical range of 80% to 125% of 3.8×10 ⁴ μg·h/mL.
The pharmaceutical composition. The pharmaceutical composition according to any one of claims 1 to 8,
Administered intravenously in multiple doses at intervals ranging from once per week to once every two weeks;
The pharmaceutical composition. A therapeutic agent for Crohn's disease comprising an anti-fractalkine antibody,
The anti-fractalkine antibody is intravenously administered at 10 to 15 mg/kg of human body weight per dose to a human patient having a Crohn's disease activity index (CDAI) of 220 or more and less than 330, at an administration interval of once a week to once every two weeks, so that the mean trough concentration is 80 μg/mL or more;
The anti-fractalkine antibody is
the heavy chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 13 (QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYYIHWVKQAPGQGLEWIGWIYPGDGSPKFNERFKGRTTLTADKSTNTAYMLLSSLRSEDTAVYFCATGPTDGDYFDYWGQGTTVTVSS);
the light chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 14 (DIQMTQSPSSLSASVGDRVTITCRASGNIHNFLAWYQQKPGKAPKLLIYNEKTLADGVPSRFSGSGSGTDYTLTISSLQPEDFATYFCQQFWSTPYTFGGGTKVEIK);
a constant region of a human IgG2 isotype; and the human IgG2 isotype constant region Fc region comprises the mutations V234A and G237A.
The drug for treating Crohn's disease.