Treatment of IgE-mediated allergic diseases

Abstract

A method for treating subjects with immunoglobulin E (IgE)-related conditions using antibodies capable of binding to the Cεmx domain of membrane-bound IgE. The subjects may be administered at least two doses of the antibody, with the two doses at least three months apart.

Description

[0001] Related Applications

[0002] This application claims the benefit of the filing date of U.S. Provisional Application Serial No. 62 / 579,416, filed on October 31, 2017, under 35 U.S.C. §119, the entire contents of which are incorporated herein by reference. Background Art

[0003] IgE plays a central role in mediating type I hypersensitivity reactions that cause allergic diseases, including allergic asthma, allergic rhinitis, atopic dermatitis, etc. Allergic reactions are responses of the immune system to harmless environmental substances, such as dust mites, tree and grass pollens, certain foods and drugs, and stings of bees and fire ants. In such reactions, the binding of allergens to IgE on the surface of basophils and mast cells causes IgE cross-linking and leads to potential receptor aggregation of IgE.Fc, type I IgE.Fc receptors, or FcεRI. This receptor aggregation then activates signal transduction pathways, leading to exocytosis of particles and release of pharmacological mediators such as histamine, leukotrienes, tryptases, cytokines, and chemokines. The release of those mediators by mast cells and basophils can cause various pathological manifestations of allergies.

[0004] There are two types of IgE molecules, free (or soluble) IgE and membrane-bound IgE (mIgE). Free IgE molecules circulate in the blood and interstitial fluid. mIgE is expressed on the surface of B lymphoblasts and memory B cells. It is believed that targeting mIgE can effectively inhibit the production of antigen-specific IgE and thus inhibit IgE-mediated immune responses. Summary of the invention

[0005] The present disclosure is based on the unexpected discovery that a single dose of FB825, an antibody targeting the CεmX domain of mIgE on human B lymphocytes, successfully reduced total IgE levels in human subjects for at least three months.

[0006] Therefore, one aspect of the present disclosure provides a method for treating a disorder associated with IgE, the method comprising administering to a subject in need thereof a first dose of an antibody that binds to the Cεmx domain of membrane-bound IgE; and administering to the subject a second dose of the antibody. The second dose is administered at least 8 weeks (e.g., at least 10 weeks, 12 weeks, or 3 months) after the first dose.

[0007] In any of the methods described herein, the first dose, the second dose, or both can range from 0.5 mg / kg to 15 mg / kg (e.g., 1 mg / kg to 15 mg / kg). For example, the first dose, the second dose, or both can range from 1 mg / kg to 8 mg / kg (e.g., 1.5 mg / kg to 10 mg / kg). The first dose, the second dose, or both can be administered by intravenous injection.

[0008] The subject treated by the methods described herein can be a human patient suffering from or suspected of suffering from an IgE-related disorder (e.g., allergic asthma, allergic rhinitis, hyper-IgE syndrome, or atopic dermatitis). In some embodiments, the disorder is cold-induced urticaria, chronic urticaria, cholinergic urticaria, chronic sinusitis, systemic mastocytosis, cutaneous mastocytosis, allergic bronchopulmonary aspergillosis, recurrent idiopathic angioedema and interstitial cystitis, eosinophil-related gastrointestinal disorders, food allergies, or drug allergies.

[0009] On the other hand, the present disclosure provides a method for treating atopic dermatitis, the method comprising administering a first dose of an antibody that binds to the Cεmx domain of membrane-bound IgE to a subject in need thereof; wherein the first dose is about 1 mg / kg to 10 mg / kg (e.g., 3 mg / kg to 8 mg / kg, such as 5 mg / kg). If at the second dose, the change in the total IgE level in the subject relative to the total IgE level before the first dose is less than 50%, the method further comprises administering a second dose of the antibody to the subject about 3 months after the first dose. In some cases, the second dose may be the same as the first dose, such as 5 mg / kg. The first dose of the antibody, the second dose of the antibody, or both may be administered by intravenous infusion.

[0010] In any of the above methods, a moisturizer is applied to the subject at least twice a day for at least seven consecutive days before the first dose. Alternatively or additionally, the method may further include applying a topical corticosteroid to the subject. In some cases, a topical corticosteroid is applied daily to active lesions. Such topical corticosteroids may be 0.05% fluticasone propionate cream, 0.1% monetasone furoate cream, 0.06% betamethasone valerate, or 1% hydrocortisone ointment. In other cases, the topical corticosteroid may be 0.05% fluticasone acetonide cream, 0.25% desoximetasone ointment, or 0.05% clobetasol propionate ointment.

[0011] In some embodiments, the subject has not been treated with topical tacrolimus, topical pimecrolimus, systemic corticosteroids, leukotriene inhibitors, allergen immunotherapy, treatment involving immunosuppressants or immunomodulators, vaccine treatment, treatment involving traditional Chinese medicine, surgical procedures, ultraviolet procedures, or tanning.

[0012] In any of the treatment methods described herein, the anti-CεmX antibodies described herein may bind to the mIgE fragment GLAGGSAQSQRAPDRVL (SEQ ID NO: 1) or the mIgE fragment GLAGGSAQSQRA (SEQ ID NO: 7). In some cases, the antibody binds to the same epitope as antibody 4B12 (FB825), or competes with antibody FB825 for binding to the Cεmx domain of membrane-bound IgE. In some examples, the antibody comprises the same heavy chain complementary determining region as antibody FB825; and / or the same light chain complementary determining region as antibody FB825. Such antibodies may be humanized antibodies of 4B12 (eg, FB825). The antibody may comprise a heavy chain variable region having an amino acid sequence of SEQ ID NO: 2, and / or a light chain variable region having an amino acid sequence of SEQ ID NO: 3. Any antibody used in any of the methods described herein may be a full-length antibody or an antigen-binding fragment thereof. The antibody may be a human antibody, a humanized antibody, a chimeric antibody, or a single-chain antibody.

[0013] In any of the methods described herein, the anti-CεmX antibody can be formulated into a pharmaceutical composition comprising an antibody, a buffer (e.g., a buffer comprising an amino acid such as histidine), a salt (e.g., sodium chloride), and a nonionic surfactant (e.g., polysorbate 80). In some embodiments, the pharmaceutical composition is an aqueous solution with a pH of 5 to 8. In some examples, the antibody in the pharmaceutical composition is about 10 mg / ml to 30 mg / ml (e.g., about 20 mg / ml), the concentration of the histidine buffer is about 10-30 mM (e.g., about 20 mM), the concentration of sodium chloride is about 120-160 mM (e.g., about 140 mM), and the concentration of polysorbate 80 is about 0.01-0.03% (e.g., about 0.02%). Any pharmaceutical composition described herein is also within the scope of the present disclosure.

[0014] On the other hand, provided herein is an aqueous formulation comprising any one of the following: an anti-CεmX antibody described herein (e.g., FB825 or a functional variant thereof) at a concentration of about 10 mg / ml to 30 mg / ml, a buffer containing an amino acid (e.g., histidine) at a concentration of about 10-30 mM, a salt (e.g., sodium chloride) at a concentration of about 120-160 mM, and a nonionic surfactant (e.g., polysorbate 80) at a concentration of about 0.01-0.03%, wherein the pH of the aqueous formulation is about 5-8. In one example, the concentration of the aqueous formulation comprising the antibody is about 20 mg / ml, the concentration of the histidine buffer is about 20 mM, the concentration of the sodium chloride is about 140 mM, and the concentration of the polysorbate 80 is about 0.02%, wherein the pH of the aqueous formulation is about 6.5.

[0015] Also within the scope of the present disclosure are (i) a pharmaceutical composition for treating an IgE-related disorder as described herein, wherein the pharmaceutical composition comprises an anti-CεmX antibody and a pharmaceutically acceptable carrier, and wherein the pharmaceutical composition is administered to a subject in need of treatment for at least two doses, which is at least 8 weeks (e.g., at least 10 weeks, 12 weeks or 3 months apart, or 12 weeks to 6 months apart); and (ii) use of an anti-CεmX antibody in the preparation of a medicament for treating an IgE-related disorder, wherein the medicament can be administered to a subject in need of treatment for at least two doses, which are at least three months apart.

[0016] The details of one or more embodiments of the present invention are set forth in the following description. Other features or advantages of the present invention will become apparent from the following drawings and detailed description of several embodiments, and also form the appended claims. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 The mean (±SD) change from baseline in total IgE over time in human subjects treated with a single dose of FB825 is shown. Total IgE was determined from blood samples. Baseline was defined as the last non-missed assessment (including repeated and unplanned assessments) before study drug administration.

[0019] Figure 2 Is a graphic representation showing the mean (± SD) percent change from baseline in total IgE over time in human subjects treated with a single dose of FB825. Total IgE levels were determined in blood samples obtained from human subjects. Baseline is defined as the last non-missed assessment (including repeated and unplanned assessments) before study drug administration.

[0020] Figure 3 is a graphic representation showing the mean (± SD) change in anti-drug antibodies (ADA) relative to baseline over time in human subjects treated with a single dose of FB825. ADA was measured in blood samples obtained from subjects. Baseline was defined as the last non-missed assessment (including repeated and unplanned assessments) before study drug administration.

[0021] Figure 4 V values ​​of monoclonal antibody 4B12 and monoclonal antibody FB825 (which is a humanized 4B12 antibody) are shown. H and V L Amino acid sequence alignment of the two antibodies. The differences between the two antibodies are highlighted. H and V L The complementarity determining regions (CDRs) are in bold and underlined. H : SEQ ID NO: 4. V of 4B12 L: SEQ ID NO: 5. V of FB825 H : SEQ ID NO: 2. V of FB825 L :SEQ ID NO:3.

[0022] Figure 5 is a graphical illustration showing the percent change from baseline in the Eczema Area and Severity Index (EASI) on Days 8, 15, 29, 57, 85, 92, 99, 113, 141, and 169 in an open-label exploratory study evaluating the safety and efficacy of FB825 in adults with atopic dermatitis.

[0023] Figure 6 is a graphical illustration showing the percent change from baseline in Investigator's Global Assessment (IGA) on Days 8, 15, 29, 57, 85, 92, 99, 113, 141, and 169 in an open-label exploratory study evaluating the safety and efficacy of FB825 in adults with atopic dermatitis.

[0024] Figure 7 is a graphical illustration showing the percent change from baseline in the Severity Score of Atopic Dermatitis Index (SCORAD) on Days 8, 15, 29, 57, 85, 92, 99, 113, 141, and 169 in an open-label exploratory study evaluating the safety and efficacy of FB825 in adults with atopic dermatitis.

[0025] Figure 8 is a graphical illustration showing the percent change from baseline in the itch visual analog scale (VAS) on Days 8, 15, 29, 57, 85, 92, 99, 113, 141, and 169 in an open-label exploratory study evaluating the safety and efficacy of FB825 in adults with atopic dermatitis. DETAILED DESCRIPTION OF THE INVENTION

[0027] In atopic individuals who are at increased risk of developing allergies, the concentration of IgE in the circulation system can reach more than 10 times the normal level. The concentration of allergen-specific IgE antibodies is closely related to clinical symptoms and can be more than 1000 times that of healthy individuals in patients with allergic diseases. Immunoglobulin E sensitizes effector cells such as basophils, mast cells, and activated eosinophils by occupying the high-affinity IgE receptor FcεRI on which they are expressed. In type I hypersensitivity reactions, allergens cross-link with IgE molecules bound by FcεRI and subsequently trigger effector cell degranulation, thereby releasing pro-inflammatory mediators such as histamine and leukotrienes. The IgE-mediated allergic pathway that produces mediator-related allergic symptoms initiates immune activity locally or systemically. Basophils and mast cells also release a wide range of inflammatory cytokines and chemokines, which not only directly lead to clinical symptoms, but also activate and recruit various cell types to enhance the inflammatory state. Therefore, anti-IgE therapy can weaken both IgE-mediated pathways and inflammatory diseases.

[0028] Described herein are anti-CεmX antibodies that can be used to reduce total IgE levels and thus treat Ig-E mediated disorders. Such antibodies can be administered to a subject in need of treatment in at least two doses that can be spaced at least 8 weeks apart (e.g., 10 weeks, 12 weeks, or 3 months).

[0029] Antibodies capable of binding to the CεmX domain of membrane-bound IgE

[0030] CεmX is a 52 amino acid segment located between the CH4 domain and the C-terminal membrane anchor segment of the human membrane-bound ε chain (mε). The amino acid sequence of an exemplary CεmX fragment of human mIgE is provided below (SEQ ID NO: 6):

[0031] GLAGGSAQSQ RAPDRVLCHS GQQQGLPRAA GGSVPHPRCH CGAGRADWPG PP

[0032] The antibodies described herein can bind to the CεmX domain of mIgE, such as mIgE expressed on the surface of B cells. Such antibodies can induce cell death of B cells expressing mIgE via, for example, antibody-dependent cellular cytotoxicity and / or apoptosis, thereby eliminating B cells, which will result in reduced production of free IgE. Therefore, the anti-CεmX antibodies described herein can reduce total IgE levels in subjects (e.g., human patients) treated with antibodies.

[0033] Antibodies (used interchangeably in the plural) are immunoglobulin molecules that are capable of specifically binding to targets such as carbohydrates, polynucleotides, lipids, polypeptides, etc., through at least one antigen recognition site located in the variable region of the immunoglobulin molecule. As used herein, the term "antibody" encompasses not only complete (i.e., full-length) polyclonal or monoclonal antibodies, but also antigen-binding fragments thereof (such as Fab, Fab', F(ab')2, Fv), single chains (scFv), mutants thereof, fusion proteins comprising antibody portions, humanized antibodies, chimeric antibodies, diabodies, linear antibodies, single-chain antibodies, multispecific antibodies (e.g., bispecific antibodies), and any other modified configurations of immunoglobulin molecules comprising antigen recognition sites of desired specificity, including glycosylation variants of antibodies, amino acid sequence variants of antibodies, and covalently modified antibodies. Antibodies include antibodies of any class, such as IgD, IgE, IgG, IgA, or IgM (or their subclasses), and antibodies do not have to be of any particular class. Immunoglobulins can be assigned to different classes based on the antibody amino acid sequence of their heavy chain constant domains. There are five major classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, and some of these can be further divided into subclasses (isotypes), such as IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2. The heavy chain constant domains corresponding to the different classes of immunoglobulins are called α, δ, ε, γ, and μ, respectively. The subunit structures and three-dimensional configurations of different classes of immunoglobulins are well known.

[0034] The antibodies used in the methods described herein can be of murine, rat, human, or any other origin (including chimeric or humanized antibodies).

[0035] Any antibody described herein may be monoclonal or polyclonal. "Monoclonal antibody" refers to a homogenous antibody population, and "polyclonal antibody" refers to a heterogenous antibody population. These two terms do not limit the source of the antibody or how it is prepared.

[0036] In one example, the antibody used in the methods described herein is a humanized antibody.Humanized antibodies refer to the form of non-human (such as mouse) antibodies, which are specific chimeric immunoglobulins, immunoglobulin chains or their antigen-binding fragments, which contain the minimum sequence derived from non-human immunoglobulins. In most cases, humanized antibodies are human immunoglobulins (receptor antibodies), wherein the residues of the complementary determining regions (CDRs) of the receptor are replaced by CDR residues from non-human species (donor antibodies) (such as mice, rats or rabbits) with the required specificity, affinity and ability. In some cases, the Fv framework region (FR) residues of human immunoglobulins are replaced by corresponding non-human residues. In addition, humanized antibodies may be included in the receptor antibodies or the CDR or framework sequences imported, but are not found but are included to further improve and optimize the antibody performance residues. Generally, humanized antibodies will include at least one and usually two variable domains substantially all, wherein all or substantially all of the CDR regions correspond to those of non-human immunoglobulins, and all or substantially all of the FR domains are those of human immunoglobulin consensus sequences. Humanized antibodies will also optimally comprise at least a portion of a constant region or domain (Fc) of an immunoglobulin (typically a human immunoglobulin). The antibody may have an Fc region modified as described in WO 99 / 58572. Other forms of humanized antibodies have one or more CDRs (one, two, three, four, five, six) that are altered relative to the original antibody, also referred to as one or more CDRs "derived from" one or more CDRs from the original antibody. Humanized antibodies may also involve affinity maturation.

[0037] In another example, the antibody described herein is a chimeric antibody, which may include a heavy constant region and a light constant region from a human antibody. A chimeric antibody refers to an antibody having a variable region from a first species or a portion of a variable region and a constant region from a second species. Typically, in these chimeric antibodies, the variable regions of light and heavy chains simulate the variable regions of antibodies derived from a mammalian species (e.g., non-human mammals, such as mice, rabbits and rats), while the constant region is homologous to the sequence in the antibody derived from another mammal such as the human race. In some embodiments, amino acid modifications may be performed in the variable region and / or constant region.

[0038] In some examples, the antibodies disclosed herein specifically bind to the Cεmx domain of membrane-bound IgE that can be expressed on the surface of B cells. Antibodies that "specifically bind" (used interchangeably herein) to a target or epitope are well-known terms in the art, and methods for determining such specific binding are also well-known in the art. A molecule is considered to exhibit "specific binding" if it reacts or associates with a specific target antigen more frequently, more rapidly, for a longer duration, and / or with greater affinity. If an antibody binds with greater affinity, avidity, faster, and / or higher duration than it binds to other substances, it "specifically binds to" the target antigen. For example, an antibody that specifically (or preferentially) binds to a CεmX domain epitope is an antibody that binds to the CεmX domain epitope with greater affinity, avidity, faster, and / or higher duration than it binds to other CεmX domain epitopes or non-CεmX domain epitopes. It can also be understood by reading this definition that, for example, an antibody that specifically binds to a first target antigen may or may not specifically bind or preferentially bind to a second target antigen. Thus, "specific binding" or "preferential binding" does not necessarily require (although it can include) exclusive binding. Typically, but not necessarily, reference to binding means preferential binding.

[0039] The binding affinity of the anti-CεmX antibodies described herein may be less than about 100 nM, such as less than about 50 nM, about 10 nM, about 1 nM, about 500 pM, about 100 pM, or about 50 pM to about 2 pM. The binding affinity may be expressed as K D or dissociation constant, and increased binding affinity corresponds to a decreased K D One method for determining the binding affinity of an antibody to CεmX is by measuring the binding affinity of a monofunctional Fab fragment of the antibody. To obtain a monofunctional Fab fragment, the antibody (e.g., IgG) can be cleaved with papain or recombinantly expressed. The affinity of the anti-CεmX Fab fragment of the antibody can be determined by surface plasmon resonance (BIAcore3000TM surface plasmon resonance (SPR) system, BIAcore, INC, Piscaway NJ). The kinetic association rate (k on ) and dissociation rate (k off ) (usually measured at 25°C); and the equilibrium dissociation constant (K D ) value according to k off / k on calculate.

[0040] In some embodiments, the antibody binds to the CεmX domain of human IgE and does not significantly bind to IgE from other mammalian species. In some embodiments, the antibody binds to human IgE and one or more IgEs from another mammalian species. The one or more epitopes bound by the antibody may be continuous or discontinuous.

[0041] In some embodiments, the anti-CεmX antibodies described herein bind to the N-terminal portion of the CεmX domain, such as GLAGGSAQSQRAPDRVL (SEQ ID NO: 1) or GLAGGSAQSQRA (SEQ ID NO: 7). Such antibodies may have similar Figure 4 The anti-CεmX antibody may be a humanized antibody of 4B12 (e.g., FB825). In some examples, the anti-CεmX antibody used in the methods described herein is FB825, which is a humanized antibody of 4B12 (e.g., FB825). Figure 4 ) or a functional variant thereof. See also U.S. Patent No. 8,460,664, the relevant disclosure of which is incorporated herein by reference.

[0042] The functional variant (equivalent) of FB825 has substantially the same epitope binding specificity as FB825 and exhibits substantially similar biological activities as FB825, including the activity of eliminating B cells expressing mIgE in a subject and reducing the level of total IgE. In some embodiments, the functional variant of FB825 contains the same region / residues responsible for antigen binding as FB825, such as the same specificity determining residues in CDR or the entire CDR. In other embodiments, the functional variant of FB825 comprises V H Chain and V L chain, the V H The chain includes the corresponding V H The CDRs are at least 75% (e.g., 80%, 85%, 90%, 95% or 98%) identical to H CDR1, V H CDR2 and V H CDR3, the V L The chain includes the corresponding V H The CDRs are at least 75% (e.g., 80%, 85%, 90%, 95% or 98%) identical to L CDR1, V L CDR2 and V L CDR3. For example, a functional variant of FB825 could contain V HChain and / or V L chain, the V H Chain V H CDR compared to V H CDR area (total V H CDR1, CDR2 and / or CDR3) contain up to 5 (e.g., 1, 2, 3, 4 or 5) amino acid residue variations, wherein the V L Chain V H CDR compared to V L CDR area (total V L CDR1, CDR2 and / or CDR3) contain up to 5 (e.g., 1, 2, 3, 4 or 5) amino acid residue variations.

[0043] Alternatively, a functional variant of the FB825 contains a V H The V chains are at least 75% (e.g., 80%, 85%, 90%, 95% or 98%) identical H chain and with FB825's V L The V chains are at least 75% (e.g., 80%, 85%, 90%, 95% or 98%) identical L The amino acid sequence variation can only be in V H and / or V L Occurs in one or more of the framework regions.

[0044] The "percent identity" of two amino acid sequences is determined using the following algorithm: Karlin and Altschul Proc. Natl. Acad. Sci. USA 87:2264-68, 1990, as modified in Karlin and Altschul Proc. Natl. Acad. Sci. USA 90:5873-77, 1993. Such an algorithm is incorporated into the NBLAST and XBLAST programs (version 2.0) of Altschul et al. J. Mol. Biol. 215:403-10, 1990. BLAST protein searches can be performed using the XBLAST program (score = 50, word length = 3) to obtain amino acid sequences homologous to the target protein molecule. In the event that there is a gap between the two sequences, gapped BLAST can be used as described in Altschul et al., Nucleic Acids Res. 25 (17): 3389-3402, 1997. When utilizing BLAST and Gapped BLAST programs, the default parameters of the respective programs (eg, XBLAST and NBLAST) can be used.

[0045] Antibody preparation

[0046] Antibodies capable of binding to the CεmX domain of membrane-bound IgE as described herein can be prepared by any method known in the art. See, for example, Harlow and Lane, (1988) Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory, New York.

[0047] In some embodiments, antibodies specific for a target antigen (e.g., the CεmX domain of mIgE (such as human mIgE)) can be prepared by conventional hybridoma technology. Full-length target antigens or fragments thereof, optionally coupled to carrier proteins such as KLH, can be used to immunize host animals to produce antibodies that bind to this antigen. As further described herein, the immunization pathways and schedules of the host animals are generally consistent with established and conventional antibody stimulation and production techniques. General techniques for producing mouse, humanized and human antibodies are known in the art and are described herein. It is contemplated that any mammalian subject, including humans or cells producing antibodies therefrom, can be manipulated as a basis for mammalian production, including human hybridoma cell lines. Typically, a host animal is inoculated intraperitoneally, intramuscularly, orally, subcutaneously, plantarly and / or intradermally with a certain amount of an immunogen, including as described herein.

[0048] Hybridomas can be prepared from lymphocytes and immortalized myeloma cells using Kohler, B. and Milstein, C. (1975) Nature 256: 495-497 or conventional somatic cell hybridization techniques as modified by Buck, DW, et al., In Vitro, 18: 377-381 (1982). Available myeloma cell lines that can be used for hybridization include, but are not limited to, X63-Ag8.653 and those from Salk Institute, Cell Distribution Center, San Diego, Calif., USA. Typically, the technology involves fusing myeloma cells and lymphoid cells using fusogens such as polyethylene glycol or by electrical means well known to those skilled in the art. After fusion, the cells are separated from the fusion medium and grown in a selective growth medium (such as hypoxanthine-aminopterin-thymidine (HAT) medium) to eliminate unhybridized parental cells. Any culture medium described herein supplemented with or without serum can be used to culture hybridomas that secrete monoclonal antibodies. As another alternative to cell fusion technology, EBV immortalized B cells can be used to produce anti-CεmX monoclonal antibodies of the present invention. If necessary, the hybridoma is expanded and subcloned, and the anti-immunogen activity of the supernatant is determined by conventional immunoassay procedures (eg, radioimmunoassay, enzyme immunoassay or fluorescent immunoassay).

[0049] Hybridomas that can be used as sources of antibodies encompass all derivatives and progeny cells of parent hybridomas that produce monoclonal antibodies capable of binding to the CεmX domain. Hybridomas that produce such antibodies can be grown in vitro or in vivo using known procedures. If necessary, monoclonal antibodies can be separated from culture media or body fluids by conventional immunoglobulin purification methods, such as ammonium sulfate precipitation, gel electrophoresis, dialysis, chromatography, and ultrafiltration. If there is undesirable activity, it can be removed, for example, by running the preparation on an adsorbent made of an immunogen attached to a solid phase and eluting or releasing the desired antibody from the immunogen. Antibody (e.g., monoclonal antibody) populations can be generated by immunizing a host animal with an antigen of interest or a fragment containing a target amino acid sequence conjugated to a protein that is immunogenic in the species to be immunized (e.g., keyhole limpet hemocyanin, serum albumin, bovine thyroglobulin, or soybean trypsin inhibitor), using a bifunctional agent or a derivatizing agent (e.g., maleimidobenzoyl sulfosuccinimide ester (coupled through cysteine ​​residues), N-hydroxysuccinimide (through lysine residues), glutaraldehyde, succinic anhydride, SOCl, or R1N=C=NR), wherein R and R1 are different alkyl groups.

[0050] If necessary, the target antibody (monoclonal or polyclonal) (as produced by hybridoma) can be sequenced, and then the polynucleotide sequence can be cloned into a vector to express or propagate. The sequence encoding the target antibody can be maintained in a vector in a host cell, and then the host cell can be amplified and frozen for future use. In an alternative, the polynucleotide sequence can be used for genetic manipulation with "humanized" antibodies or to improve the affinity (affinity maturation) or other characteristics of the antibody. For example, if the antibody is used in the clinical trial and treatment of the mankind, the constant region can be engineered to be more similar to the human constant region to avoid immune response. It can be expected that the antibody sequence is genetically manipulated to obtain a greater affinity to the target antigen and to reduce the greater effect of total IgE. It is obvious to those skilled in the art that one or more polynucleotide changes can be carried out on the antibody, and still maintain its binding specificity with the target antigen.

[0051] In other embodiments, fully human antibodies can be obtained by using commercially available mice that have been engineered to express specific human immunoglobulin proteins. Transgenic animals designed to produce more desirable (e.g., fully human antibodies) or more robust immune responses can also be used to generate humanized or human antibodies. Examples of such technologies are XenomouseRTM from Amgen, Inc. (Fremont, Calif.) and HuMAb-MouseRTM and TC MouseTM from Medarex, Inc. (Princeton, NJ). In another alternative, antibodies can be recombinantly prepared by phage display technology. See, for example, U.S. Patent Nos. 5,565,332; 5,580,717; 5,733,743; and 6,265,150; and Winter et al., (1994) Annu.Rev.Immunol.12:433-455. Alternatively, phage display technology (McCafferty et al., (1990) Nature 348:552-553) can be used to produce human antibodies and antibody fragments in vitro from immunoglobulin variable (V) domain gene repertoires from unimmunized donors.

[0052] Antigen-binding fragments of intact antibodies (full-length antibodies) can be prepared by conventional methods. For example, F(ab')2 fragments can be produced by digesting antibody molecules with pepsin, and Fab fragments can be generated by reducing the disulfide bridges of the F(ab')2 fragments.

[0053] Antibodies through genetic engineering, such as humanized antibodies, chimeric antibodies, single-chain antibodies and bispecific antibodies can be produced via conventional recombinant techniques. In one example, conventional methods (such as, by using oligonucleotide probes that can specifically bind to the heavy chain and light chain of the monoclonal antibody encoding the monoclonal antibody) can be easily separated to encode the DNA of the monoclonal antibody specific to the target antigen, and sequenced. Hybridoma cells are used as the preferred source of such DNA. Once separated, DNA can be placed in one or more expression vectors, which are then transfected into host cells, such as Escherichia coli cells, monkey COS cells, Chinese hamster ovary (CHO) cells or myeloma cells that do not otherwise produce immunoglobulin proteins, to obtain the synthesis of monoclonal antibodies in recombinant host cells. See, such as PCT Publication No. WO 87 / 04462. The DNA can then be modified, for example, by replacing the homologous mouse sequences with coding sequences for human heavy and light chain constant domains, Morrison et al., (1984) Proc. Nat. Acad. Sci. 81: 6851, or by covalently linking the immunoglobulin coding sequence to all or part of a non-immunoglobulin polypeptide coding sequence. In this way, genetically engineered antibodies, such as "chimeric" or "hybrid" antibodies, can be prepared to have target antigen binding specificity.

[0054] Techniques developed for the production of "chimeric antibodies" are well known in the art. See, e.g., Morrison et al. (1984) Proc. Natl. Acad. Sci. USA 81, 6851; Neuberger et al. (1984) Nature 312, 604; and Takeda et al. (1984) Nature 314: 452.

[0055] Methods for constructing humanized antibodies are also well known in the art. See, e.g., Queen et al., Proc. Natl. Acad. Sci. USA, 86:10029-10033 (1989). In one example, the V of a parent non-human antibody is modified according to methods known in the art. H and V L The variable regions of the parent V were subjected to three-dimensional molecular modeling analysis. Next, the same molecular modeling analysis was used to identify the framework amino acid residues predicted to be important for forming the correct CDR structure. H and V L Sequences were used as search terms, and parent V H and V L The human V H and V L Receptor gene.

[0056] The CDR region in the selected human receptor gene can be replaced by the CDR region from the parent non-human antibody or its functional variant. If necessary, the residues in the parent chain framework region (see above) predicted to be important in the interaction with the CDR region can be used to replace the corresponding residues in the human receptor gene.

[0057] Single-chain antibodies can be prepared by recombinant techniques by connecting the nucleotide sequence encoding the heavy chain variable region and the nucleotide sequence encoding the light chain variable region. Preferably, a flexible linker is incorporated between the two variable regions. Alternatively, the techniques described for producing single-chain antibodies (U.S. Patent Nos. 4,946,778 and 4,704,692) can be adapted to produce phage scFv libraries, and scFv clones specific for IgE can be identified from the library according to conventional procedures.

[0058] Methods well known in the art can be used to characterize antibodies obtained according to methods known in the art and described herein. For example, one method is to identify the epitope bound by the antigen or "epitope mapping". There are many methods in the art that can be used to map and characterize the position of epitopes on proteins, including crystal structures of antibody-antigen complexes, competitive assays, gene fragment expression assays, and assays based on synthetic peptides, such as Harlow and Lane, Using Antibodies, a Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 1999, Chapter 11. In another example, epitope mapping can be used to determine the sequence bound by the antibody. The epitope can be a linear epitope, i.e., a conformational epitope (primary structure linear sequence) formed by the three-dimensional interaction of amino acids contained in a single amino acid segment, or may not necessarily be contained in a single segment. Peptides of different lengths (e.g., at least 4-6 amino acids long) can be separated or synthesized (e.g., recombined) and used for binding assays with antibodies. In another example, the epitope bound to the antibody can be determined in a systematic screening by using overlapping peptides derived from the target antigen sequence and determining the binding of the antibody. According to the gene fragment expression assay, the open reading frame encoding the target antigen is randomly or fragmented by a specific genetic construction, and the reactivity of the antigen expression fragment with the antibody to be tested is determined. The gene fragment can be produced, for example, by PCR, and then transcribed and translated into protein in vitro in the presence of radioactive amino acids. The combination of the antibody and the radiolabeled antigen fragment is then determined by immunoprecipitation and gel electrophoresis. Some epitopes can also be identified by using a large random peptide sequence library (phage library) displayed on the surface of phage particles. Alternatively, the combination of the limited library of overlapping peptide fragments and the test antibody can be tested in a simple binding assay. In another example, mutagenesis, domain exchange experiments and alanine scanning mutagenesis of the antigen binding domain can be carried out to identify the residues required, sufficient and / or necessary for the epitope to bind. For example, a mutant of the target antigen can be used to carry out a domain exchange experiment, in which each fragment of the IgE polypeptide has been replaced (exchanged) by a sequence from a closely related but different antigenic protein (such as another member of the immunoglobulin family). By assessing the combination of the antibody and the mutant immunoglobulin, the importance of the specific antigen fragment to the antibody binding can be assessed.

[0059] Alternatively, competition assays can be performed using other antibodies known to bind to the same antigen to determine whether the antibody binds to the same epitope as the other antibodies. Competition assays are well known to those skilled in the art.

[0060] Pharmaceutical composition

[0061] One or more of the above-mentioned anti-CεmX antibodies can be mixed with a pharmaceutically acceptable carrier (excipient) (including a buffer) to form a pharmaceutical composition for treating a disorder associated with IgE. "Acceptable" means that the carrier must be compatible with the active ingredient of the composition (and preferably capable of stabilizing the active ingredient) and harmless to the subject to be treated. Pharmaceutically acceptable excipients (carriers), including buffers, are well known in the art. See, e.g., Remington: The Science and Practice of Pharmacy 20th edition (2000) Lippincott Williams and Wilkins, ed. KE Hoover. In one example, the pharmaceutical composition contains more than one anti-CεmX antibody that recognizes different epitopes of the target antigen.

[0062] The pharmaceutical composition to be used in the method of the present invention may include a pharmaceutically acceptable carrier, excipient or stabilizer in the form of a lyophilized formulation or an aqueous solution. Remington: The Science and Practice of Pharmacy 20th Edition (2000) Lippincott Williams and Wilkins, ed. KE Hoover. Acceptable carriers, excipients or stabilizers are nontoxic to the recipient at the dosages and concentrations used, and may include buffers such as phosphates, citrates and other organic acids; antioxidants including ascorbic acid and methionine; preservatives (such as octadecyldimethylbenzyl ammonium chloride; hexamethylammonium chloride; benzalkonium chloride, benzethonium chloride; phenol, butyl or benzyl alcohol; alkyl parabens such as methyl or propyl paraben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecular weight (less than about 10 residues) polypeptides; proteins such as serum albumin, gelatin or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine or lysine; monosaccharides, disaccharides and other carbohydrates including glucose, mannose or dextran; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose or sorbitol; salt-forming counterions such as sodium; metal complexes (e.g., Zn-protein complexes); and / or nonionic surfactants such as TWEEN®. TM 、PLURONICS TM Or polyethylene glycol (PEG). Pharmaceutically acceptable excipients are also described herein.

[0063] In some examples, the pharmaceutical compositions described herein include liposomes containing anti-CεmX antibodies, which can be prepared by methods known in the art, such as Epstein et al., Proc. Natl. Acad. Sci. USA 82: 3688 (1985); Hwang et al., Proc. Natl. Acad. Sci. USA 77: 4030 (1980); and U.S. Pat. Nos. 4,485,045 and 4,544,545. Liposomes with enhanced circulation time are disclosed in U.S. Pat. No. 5,013,556. Particularly useful liposomes can be produced by reverse phase evaporation with a lipid composition comprising phosphatidylcholine, cholesterol and PEG-derivatized phosphatidylethanolamine (PEG-PE). Liposomes are extruded through a filter of defined pore size to produce liposomes with a desired diameter.

[0064] Anti-CεmX antibodies can also be embedded in microcapsules prepared, for example, by coacervation techniques or by interfacial polymerization, such as hydroxymethylcellulose or gelatin-microcapsules and poly(methyl methacrylate) microcapsules in colloidal drug delivery systems (e.g., liposomes, albumin microspheres, microemulsions, nanoparticles and nanocapsules) or in macroemulsions, respectively. Such techniques are known in the art, see, e.g., Remington, The Science and Practice of Pharmacy 20th edition Mack Publishing (2000).

[0065] In other examples, the pharmaceutical compositions described herein can be formulated into sustained release forms. Suitable examples of sustained release formulations include semipermeable matrices of solid hydrophobic polymers containing antibodies, the matrix being in the form of molded articles, such as films or microcapsules. Examples of sustained release matrices include polyesters, hydrogels (e.g., poly(2-hydroxyethyl-methacrylate) or poly(vinyl alcohol), polylactic acid (U.S. Patent No. 3,773,919), copolymers of L-glutamic acid and 7-ethyl-L-glutamic acid, non-degradable ethylene-vinyl acetate, degradable lactic acid-glycolic acid copolymers (such as LUPRON DEPOT TM (injectable microspheres composed of lactic acid-glycolic acid copolymer and leuprolide acetate), sucrose acetate isobutyrate, and poly-D-(-)-3-hydroxybutyric acid.

[0066] In some embodiments, a pharmaceutical composition comprising an anti-CεmX antibody described herein (e.g., FB825 or a functional variant thereof, also as described herein) may be a liquid formulation, which may further comprise a buffer (which may comprise an amino acid, such as histidine or arginine), a salt (e.g., sodium chloride), and / or a surfactant (such as a nonionic surfactant). For example, an aqueous formulation may comprise an antibody at a concentration of about 10-30 mg / ml, a buffer comprising an amino acid (e.g., histidine or arginine) at a concentration of about 10-30 mM, a surfactant (such as polysorbate 80) at a concentration of about 0.01-0.03%, and / or sodium chloride at a concentration of about 120-160 mM. Such an aqueous formulation may have a pH of about 5-8. In a specific example, an aqueous formulation can include FB825 antibody at a concentration of about 20 mg / ml, L-histidine at a concentration of about 20 mM, sodium chloride at a concentration of about 140 mM, polysorbate 80 at a concentration of about 0.02%, and a pH of about 6.5.

[0067] The term "about" or "approximately" means that a particular value is within an acceptable error range as determined by one of ordinary skill in the art, which will depend in part on how the value is measured or determined, i.e., the limitations of the measurement system. For example, according to the practice in the art, "about" may mean within an acceptable standard deviation. Alternatively, "about" may mean a range of up to ±20%, preferably up to ±10%, more preferably up to ±5%, and still more preferably up to ±1% of a given value. Alternatively, particularly with respect to biological systems or processes, the term may mean within the order of magnitude of a numerical value, preferably within twice the numerical value. Where a particular value is described in the present application and claims, the term "about" is implicit, unless otherwise stated, and in this context means that the particular value is within an acceptable error range.

[0068] Pharmaceutical compositions for in vivo administration must be sterile. This is easily achieved, for example, by filtration through a sterile filtration membrane. The therapeutic antibody composition is usually placed in a container with a sterile inlet, for example, an intravenous solution bag or vial with a stopper that can be pierced by a hypodermic injection needle.

[0069] The pharmaceutical compositions described herein may be in unit dosage form such as tablets, pills, capsules, powders, granules, solutions or suspensions, or suppositories, for oral, parenteral or rectal administration, or administration by inhalation or insufflation.

[0070] In order to prepare solid compositions, such as tablets, the main active ingredient can be mixed with a pharmaceutical carrier (such as conventional tableting ingredients, such as corn starch, lactose, sucrose, sorbitol, talc, stearic acid, magnesium stearate, dicalcium phosphate or gum) and other pharmaceutical diluents (such as water) to form a solid preformulated composition containing a homogeneous mixture of the compound of the present invention or its nontoxic pharmaceutically acceptable salt. When these preformulated compositions are referred to as homogeneous, it means that the active ingredient is evenly dispersed throughout the composition, so that the composition can be easily subdivided into equivalent unit dosage forms, such as tablets, pills and capsules. The solid preformulated composition is then subdivided into unit dosage forms of the above type, which contain 0.1 to about 500 mg of the active ingredient of the present invention. The tablets or pills of the novel composition can be coated or otherwise compounded to provide a dosage form that provides an advantage of prolonged action. For example, a tablet or pill can include an internal dose and an external dose component, the latter being an encapsulated form on the former. The two components may be separated by an enteric layer which serves to resist disintegration in the stomach and permit the inner component to pass intact into the duodenum or to be delayed in release. A variety of materials can be used for such enteric layers or coatings, such materials including a variety of polymeric acids and mixtures of polymeric acids with materials such as shellac, cetyl alcohol, and cellulose acetate.

[0071] Suitable surfactants include in particular nonionic agents such as polyoxyethylene sorbitan (eg, Tween TM 20, 40, 60, 80 or 85) and other sorbitans (e.g., Span TM 20, 40, 60, 80 or 85). Compositions with surfactants will conveniently contain 0.05 to 5% surfactant, and may be 0.1 to 2.5%. It will be appreciated that other ingredients may be added if necessary, such as mannitol or other pharmaceutically acceptable vehicles.

[0072] Commercially available fat emulsions such as Intralipid TM , Liposyn TM Infonutrol TM 、Lipofundin TM Lipiphysan TM To prepare a suitable emulsion. The active ingredient can be dissolved in a premixed emulsion composition, or alternatively can be dissolved in an oil (e.g., soybean oil, safflower oil, cottonseed oil, sesame oil, corn oil or almond oil) and an emulsion formed after mixing with a phospholipid (e.g., lecithin, soybean lecithin or soybean lecithin) and water. It should be understood that other ingredients, such as glycerol or glucose, can be added to adjust the tonicity of the emulsion. Suitable emulsions will typically contain up to 20% oil, for example 5 to 20%.

[0073] The emulsion composition can be prepared by combining the anti-CεmX antibody with Intralipid TM Or those prepared by mixing its components (soybean oil, lecithin, glycerin and water).

[0074] Pharmaceutical compositions for inhalation or insufflation include solutions and suspensions in pharmaceutically acceptable, aqueous or organic solvents, or mixtures thereof, and powders. Liquid or solid compositions may contain suitable pharmaceutically acceptable excipients as described above. In some embodiments, the compositions are administered by the oral or nasal respiratory route for local or systemic effect.

[0075] Compositions in preferably sterile pharmaceutically acceptable solvents may be aerosolized by the use of gases. Aerosolized solutions may be breathed directly from the aerosolizing device, or the aerosolizing device may be attached to a mask, tent, or intermittent positive pressure breathing machine. Solutions, suspensions, or powder compositions may be administered orally or nasally, preferably from a device that delivers the formulation in an appropriate manner.

[0076] Use of anti-CεmX antibodies for treating gE-related disorders

[0077] In order to implement the methods disclosed herein, an effective amount of the above-mentioned pharmaceutical composition can be administered to a subject (e.g., a person) in need of treatment via a suitable route (such as intravenous administration, such as as a bolus or by continuous infusion over a period of time, by intramuscular, intraperitoneal, intraspinal, subcutaneous, intraarticular, intrasynovial, intrathecal, oral, inhaled or topical routes). Commercially available nebulizers for liquid preparations, including jet nebulizers and ultrasonic nebulizers, can be used for administration. Liquid preparations can be directly atomized after reconstitution, and lyophilized powders can be atomized. Alternatively, anti-CemX antibodies can be atomized using fluorocarbon preparations and metered dose inhalers, or inhaled as lyophilized and ground powders.

[0078] The subject treated by the methods described herein can be a mammal, more preferably a human. Mammals include but are not limited to farm animals, sports animals, pets, primates, horses, dogs, cats, mice and rats. Human subjects in need of treatment can be patients with conditions associated with IgE (e.g., allergic asthma and other conditions known in the art and / or disclosed herein), in conditions associated with IgE (e.g., allergic asthma and other conditions known in the art and / or disclosed herein) risk or suspected patients with conditions associated with IgE (e.g., allergic asthma and other conditions known in the art and / or disclosed herein). It is possible to determine the subject with IgE related conditions (such as allergic asthma) by routine medical examinations, such as laboratory tests. Subjects suspected of having IgE related conditions may show one or more symptoms of conditions, such as, elevated levels of IgE and / or hyperreactivity to allergens and / or antigens. Subjects in risk of conditions can be subjects with one or more risk factors for this condition.

[0079] Exemplary IgE-associated disorders include, but are not limited to, asthma, allergic rhinitis, hyper-IgE syndrome, atopic dermatitis, cold-induced urticaria, chronic urticaria, cholinergic urticaria, chronic sinusitis, systemic mastocytosis, cutaneous mastocytosis, allergic bronchopulmonary aspergillosis, recurrent idiopathic angioedema and interstitial cystitis, eosinophil-associated gastrointestinal disorders, food allergies, or drug allergies.

[0080] As used herein, "effective amount" refers to the amount of each active agent required to give a subject a therapeutic effect, either alone or in combination with one or more other active agents. As recognized by those skilled in the art, the effective amount varies according to the specific illness being treated, the severity of the illness, individual patient parameters (including age, physical condition, size, sex and weight), duration of treatment, the nature of concurrent therapy (if any), specific route of administration, and similar factors within the knowledge and expertise of health practitioners. These factors are well known to those of ordinary skill in the art and can only be solved by routine experiments. It is usually preferred to use the maximum dose of a single component or its combination, i.e., the highest safe dose according to reasonable medical judgment. However, it will be understood by those of ordinary skill in the art that patients may adhere to lower doses or tolerable doses for medical reasons, psychological reasons, or indeed any other reason.

[0081] Empirical considerations such as half-life will generally help determine the dosage. For example, antibodies compatible with the human immune system, such as humanized antibodies or fully human antibodies, can be used to extend the half-life of the antibody and prevent the antibody from being attacked by the host's immune system. The frequency of administration can be determined and adjusted during the course of treatment, and is usually but not necessarily based on the treatment and / or inhibition and / or improvement and / or delay of IgE-related disorders. Alternatively, a sustained continuous release formulation of an anti-CεmX antibody may be appropriate. Various preparations and devices for achieving sustained release are known in the art.

[0082] In one example, the dosage of an anti-CεmX antibody as described herein can be determined empirically in an individual who has been given one or more administrations of an anti-CεmX antibody. Increasing doses of the anti-CεmX antibody are given to the individual. To assess the efficacy of the anti-CεmX antibody, indicators of IgE-related disorders (such as IgE levels) can be followed up.

[0083] For the purposes of the present disclosure, the appropriate dose of an anti-CεmX antibody will depend on the specific anti-CεmX antibody or antibodies (or compositions thereof) employed, the type and severity of the disorder associated with IgE, whether the antibody is administered for preventive or therapeutic purposes, previous therapy, the patient's clinical history and response to the antibody, and the discretion of the attending physician. Typically, the clinician will administer an anti-CεmX antibody (such as FB825) until a dose is reached that achieves the desired result. Administration of the anti-CεmX antibody may be continuous or intermittent, for example, depending on the physiological condition of the recipient, whether the administration is therapeutic or prophylactic, and other factors known to the skilled artisan.

[0084] In some embodiments, an anti-CεmX antibody described herein (e.g., FB825) is administered to a subject in need of treatment in an amount sufficient to reduce total IgE levels by at least 20% (e.g., 30%, 40%, 50%, 60%, 70%, 80%, 90% or more).

[0085] As used herein, the term "treat" refers to applying or administering a composition comprising one or more active agents to a subject suffering from, having symptoms of, or being predisposed to an IgE-related disease, with the purpose of curing, alleviating, alleviating, relieving, altering, remedying, improving, ameliorating, or affecting the condition, disease symptoms, or predisposition to the disease.

[0086] Alleviate the disease associated with IgE and include delaying the development or progress of the disease, or reducing the severity of the disease. Alleviate the disease does not necessarily require a cure. As used herein, the development of "delaying" a disease (such as a disease associated with IgE) means to delay, hinder, slow down, slow down, stabilize and / or postpone the progress of the disease. Depending on the disease treated and / or personal medical history, this delay may have different lengths of time. "Delay" or alleviate the development of a disease or delay the onset of a disease is when compared to not using the method, reducing the possibility of developing one or more symptoms of a disease within a given time frame and / or reducing the degree of symptoms within a given time frame. Such comparisons are usually based on clinical studies, using multiple subjects that are sufficient to give statistically significant results to carry out.

[0087] "Development" or "progression" of a disease means the initial manifestation and / or subsequent progression of the disease. Standard clinical techniques well known in the art can be used to detect and assess the development of a disease. However, development can also refer to progression that may not be detected. For the purposes of this disclosure, development or progression refers to the biological course of symptoms. "Development" includes occurrence, recurrence, and onset. As used herein, "onset" or "occurrence" of a disease associated with IgE includes initial onset and / or recurrence.

[0088] To perform the methods described herein, any of the anti-Cεmx antibodies (such as FB825) can be administered to a subject (e.g., a human patient) in need of treatment by a single dose or by multiple doses, via a suitable route (e.g., intravenous infusion or subcutaneous injection). The dose range of the anti-Cεmx antibody administered per dose is about 0.5 mg / kg to about 25 mg / kg (e.g., about 1 mg / kg to about 20 mg / kg, about 5 mg / kg to about 15 mg / kg, or about 10 mg / kg to about 20 mg / kg), depending on various factors, including those described herein. For repeated administration over several days or longer, depending on the circumstances, treatment is continued until the desired symptom suppression occurs or until a therapeutic level sufficient to alleviate the disorder associated with IgE or its symptoms is reached.

[0089] Administration of an anti-CεmX antibody (e.g., FB825) may be substantially continuous over a predetermined time or may be in a series of spaced doses, such as before, during, or after the development of an IgE-related disorder. An exemplary dosing regimen includes administering a first dose of an anti-Cemx antibody (e.g., 3 mg / kg, 5 mg / kg, 10 mg / kg, 15 mg / kg, 20 mg / kg, or 25 mg / kg) to a subject in need of treatment, followed by administration of a second dose of the antibody at least 3 months (e.g., 4 months, 5 months, or 6 months) after the first dose. The dose of the second administration may be higher, the same, or lower than the first administration. Other dosage regimens may be useful depending on the pattern of pharmacokinetic attenuation that the practitioner wishes to achieve.

[0090] In some embodiments, a subject in need of treatment can be given a suitable amount (e.g., 3 mg / kg, 5 mg / kg, 10 mg / kg, 15 mg / kg, 20 mg / kg, or 25 mg / kg). The subject is then regularly monitored for symptoms that indicate an IgE-related condition, such as an allergic reaction and / or an increase in total IgE levels. When such symptoms are observed, a second dose of the antibody can be given to the subject.

[0091] Also within the scope of the present disclosure is to use any of the anti-Cεmx antibodies to prophylactically treat IgE-related disorders to reduce the risk of developing such disorders. Subjects suitable for such prophylactic treatment can be human patients with a history of IgE-related disorders and / or a family history of IgE-related disorders.

[0092] According to the type of disease to be treated or disease site, the pharmaceutical composition can be administered to the subject using conventional methods known to those of ordinary skill in the medical field. The composition can also be administered via other conventional routes, such as oral, parenteral, by inhalation spray, topically, rectal, nasal, buccal, vaginally or via implanted reservoir administration. As used herein, the term "parenteral" includes subcutaneous, intradermal, intravenous, intramuscular, intraarticular, intraarterial, intrasynovial, intrasternal, intrathecal, intralesional and intracranial injection or infusion techniques. In addition, it can be administered to the subject via injectable reservoir administration routes, such as using 1, 3, 6 months reservoir injectable or biodegradable materials and methods.

[0093] Injectable compositions may contain various carriers, such as vegetable oils, dimethylacetamide, dimethylformamide, ethyl lactate, ethyl carbonate, isopropyl myristate, ethanol and polyols (glycerol, propylene glycol, liquid polyethylene glycol, etc.). For intravenous injection, water-soluble antibodies may be administered by drip infusion, whereby a pharmaceutical preparation containing the antibody and a physiologically acceptable excipient is infused. Physiologically acceptable excipients may include, for example, 5% glucose, 0.9% saline, Ringer's solution or other suitable excipients. Intramuscular preparations, such as sterile preparations in the form of suitable soluble salts of the antibody, may be dissolved in a pharmaceutical excipient (such as water for injection, 0.9% saline or 5% glucose solution) and administered therein.

[0094] In one embodiment, the anti-CεmX antibody is administered via a site-specific or targeted local delivery technique. Examples of site-specific or targeted local delivery techniques include various implantable reservoir sources or local delivery catheters of anti-CεmX antibodies, such as infusion catheters, indwelling catheters or needle catheters, synthetic grafts, outer membrane packaging, shunts and stents or other implantable devices, site-specific carriers, direct injection or direct application. See, e.g., PCT Publication No. WO 00 / 53211 and U.S. Patent No. 5,981,568. Targeted delivery of therapeutic compositions containing antisense polynucleotides, expression vectors or subgenomic polynucleotides can also be used. Receptor-mediated DNA delivery techniques are described, for example, in Findeis et al., Trends Biotechnol. (1993) 11:202; Chiou et al., Gene Therapeutics: Methods And Applications Of Direct Gene Transfer (J. A. Wolff, ed.) (1994); Wu et al., J. Biol. Chem. (1988) 263:621; Wu et al., J. Biol. Chem. (1994) 269:542; Zenke et al., Proc. Natl. Acad. Sci. USA (1990) 87:3655; Wu et al., J. Biol. Chem. (1991) 266:338. Therapeutic compositions containing polynucleotides are administered locally in a gene therapy regimen in the range of about 100 ng to about 200 mg DNA. In some embodiments, concentrations ranging from about 500 ng to about 50 mg, about 1 μg to about 2 mg, about 5 μg to about 500 μg, and about 20 μg to about 100 μg or more of DNA may also be used during a gene therapy regimen.

[0095] The therapeutic polynucleotides and polypeptides described herein can be delivered using gene delivery vehicles. The gene delivery vehicle can be viral or non-viral in origin (see generally Jolly, Cancer Gene Therapy (1994) 1: 51; Kimura, Human Gene Therapy (1994) 5: 845; Connelly, Human Gene Therapy (1995) 1: 185; and Kaplitt, Nature Genetics (1994) 6: 148). Endogenous mammalian or heterologous promoters and / or enhancers can be used to induce the expression of such coding sequences. The expression of coding sequences can be constitutive or regulated.

[0096] Viral-based vectors for delivery of a desired polynucleotide and expression in a desired cell are well known in the art. Exemplary viral-based vectors include, but are not limited to, recombinant retroviruses (see, e.g., PCT Publication Nos. WO 90 / 07936; WO 94 / 03622; WO 93 / 25698; WO 93 / 25234; WO 93 / 11230; WO 93 / 10218; WO 91 / 02805; U.S. Pat. Nos. 5,219,740 and 4,777,127; GB Patent No. 2,200,651; and European Patent No. 0 345242), alphavirus-based vectors (e.g., Sindbis virus vectors, Semliki Forest virus (ATCC VR-67; ATCC VR-1247), Ross River virus (ATCC VR-373; ATCC VR-1246), and Venezuelan equine encephalitis virus (ATCC VR-923; ATCC VR-1250; ATCC VR 1249; ATCC VR-1250). VR-532)); and adeno-associated virus (AAV) vectors (see, e.g., PCT Publication Nos. WO 94 / 12649, WO 93 / 03769; WO 93 / 19191; WO 94 / 28938; WO 95 / 11984 and WO 95 / 00655). Administration of DNA linked to a killed adenovirus can also be employed, as described in Curiel, Hum. Gene Ther. (1992) 3:147. Non-viral delivery vehicles and methods can also be used, including but not limited to polycation condensed DNA linked or unlinked to a separate killed adenovirus (see, e.g., Curiel, Hum. Gene Ther. (1992) 3: 147); ligand-linked DNA (see, e.g., Wu, J. Biol. Chem. (1989) 264: 16985); eukaryotic cell delivery vehicle cells (see, e.g., U.S. Pat. No. 5,814,482; PCT Publication Nos. WO 95 / 07994; WO 96 / 17072; WO 95 / 30763; and WO 97 / 42338) and nucleic acid charge neutralization or fusion with cell membranes. Naked DNA can also be used. Exemplary naked DNA introduction methods are described in PCT Publication No. WO 90 / 11092 and U.S. Pat. No. 5,580,859. Liposomes that can be used as gene delivery vehicles are described in US Pat. No. 5,422,120; PCT Publication Nos. WO 95 / 13796; WO 94 / 23697; WO 91 / 14445; and European Patent No. 0524968.Additional methods are described in Philip, Mol. Cell. Biol. (1994) 14:2411, and Woffendin, Proc. Natl. Acad. Sci. (1994) 91:1581.

[0097] It is also significant that expression vectors can be used to direct the expression of any of the protein-based anti-CεmX antibodies described herein (eg, FB825). For example, other anti-CεmX antibody fragments capable of binding to CεmX and / or IgE biological activity are known in the art.

[0098] The specific dosage regimen, i.e., dosage, timing, and repetition, used in the methods described herein will depend on the specific subject and the subject's medical history. Any anti-Cmex antibody described herein can be used in combination with other agents (e.g., other agents for treating IgE-related disorders) for enhancing and / or supplementing the effectiveness of the agent.

[0099] In some embodiments, anti-CεmX antibodies as described herein, such as FB825, are used as follows to treat atopic dermatitis. Atopic dermatitis, also known as eczema, is a chronic skin disorder characterized by redness and / or itching. It is common in children, but can occur at any age. Patients in need of treatment can be identified as having one or more symptoms of atopic dermatitis by conventional medical practice, including dry skin, itching, red to brown-gray patches, small raised bumps (which may leak fluid and crust when scratched), thickened and cracked scaly skin, and / or raw sensitive skin swollen due to scratching. In some cases, the total IgE level and allergen-specific IgE level of the candidate subject can be checked via conventional practice. If the IgE level of the candidate subject (e.g., total IgE, allergen-specific IgE, or both) is higher than the normal level (representing the average IgE level in subjects of the same species (such as humans without atopic dermatitis or other allergic conditions associated with IgE)).

[0100] Via conventional routes as described herein, a first dose of antibody can be administered to a human patient in need of treatment, and the dosage range can be 3 mg / kg to 8 mg / kg. In some cases, the first dose is 5 mg / kg. After the first dose, the total IgE level of the patient can be monitored. If the IgE level at 3-4 weeks after the first dose is reduced by less than 50%, the second dose of antibody can be administered to the patient 3-4 weeks after the first dose. The second dose can be the same as the first dose, or lower than the first dose. In some cases, the first dose and the second dose are both 5 mg / kg and are administered via intravenous infusion within 1-2 hours. During the treatment process, other biomarkers indicating efficacy and / or safety can also be monitored. Such biomarkers include, but are not limited to, thymus and activation-regulated chemokines (TARC), Eotaxin-3, thymic stromal lymphopoietin (TSLP), periostin, IL-1a, IL-4, IL-5, IL-13, IL-16, IL-31, M-CSF, or a combination thereof.

[0101] The human patient treated as described above may have chronic atopic dermatitis for at least 3 years, as diagnosed by conventional medical practice, e.g., as defined by the Hannifin and Rajka criteria modified by Eishenfield, and supported by positive allergen-specific IgE. The patient may have one or more of the following features: (i) an eczema area and severity index (EASI) score greater than 14, (ii) an investigator's global assessment (IGA) score greater than 3 (out of 5), (iii) a body surface area (BSA) greater than 10%, (iv) a history of insufficient response to a stable regimen of topical corticosteroids or calcineurin inhibitors for at least one month or at least three months prior to treatment. In addition, a stable dose of an emollient may be administered to the human patient twice daily for at least 7 days prior to treatment.

[0102] In some cases, an anti-CεmX antibody as described herein (e.g., FB825) may be used in conjunction with a moisturizer (e.g., at a stable dose, such as at least twice a day) and / or a topical corticosteroid (TCS). Intermediate-acting TCS may be applied to areas with active lesions and may be switched to low-acting TCS after the lesions are controlled. If the lesions recur, treatment with intermediate-acting TCS may be resumed using a step-down method. If lesions persist or worsen after daily treatment with intermediate-acting TCS, high- or ultra-high-acting TCS may be used unless it is considered unsafe. Low-acting TCS may be used in areas with thinner skin (e.g., face, neck, tricuspid valve, genital area, or areas of skin atrophy), or in areas where continued use of intermediate-acting TCS is considered unsafe.

[0103] TCS with low, medium and high or super high efficacy are well known in the art. Exemplary medium efficacy TCS include 0.05% fluticasone propionate cream, 0.1% mometasone furoate cream or 0.06% betamethasone valerate cream. Exemplary low efficacy TCS include 1% hydrocortisone ointment. Exemplary high efficacy TCS may be 0.05% fluticasone acetonide cream or 0.25% desoximetasone ointment. Exemplary super high efficacy TCS may be 0.05% clobetasol propionate ointment.

[0104] In some cases, patients undergoing treatment as described herein are free of one or more of the following therapies: (i) topical tacrolimus and pimecrolimus, (ii) systemic treatment with corticosteroids, (iii) leukotriene inhibitors, (iv) allergen immunotherapy, (v) systemic treatment with immunosuppressants or immunomodulators (such as cyclosporine, mycophenolate mofetil, IFN-γ, azathioprine, methotrexate or biologics), (vi) live (such as attenuated) vaccines, and / or (vii) traditional Chinese medicine. Patients may also be free of any surgical procedures and / or ultraviolet procedures.

[0105] Any of the methods described herein may also include evaluating the occurrence of decreased hemoglobin, upper respiratory tract infection, urinary tract infection, or a combination thereof in the subject after the first dose. If one or more occurrences are observed, the amount of the second dose of anti-Cemx antibody (e.g., FB825) may be reduced. Alternatively, treatment may be discontinued.

[0106] Kit for treating IgE-related disorders

[0107] The present disclosure also provides kits for treating IgE-related disorders.Such kits may include one or more containers comprising an anti-Cεmx antibody as described herein (such as FB825).

[0108] In some embodiments, the kit may include instructions for use according to any of the methods described herein. The included instructions may include instructions for treating IgE-related disorders, delaying the onset of IgE-related disorders, or alleviating IgE-related disorders by administering anti-Cεmx antibodies according to any of the methods described herein. The kit may also include a description of selecting an individual suitable for treatment based on identifying whether the individual has a disorder, is suspected of having a disorder, or is at risk of a disorder. In other embodiments, the instructions include a description of administering an anti-Cεmx antibody to a subject in need of treatment to reduce the risk of developing an IgE-related disorder.

[0109] Instructions for use involving anti-Cεmx antibodies typically include information about the intended treatment dose, dosing schedule, and route of administration. The container may be a unit dose, bulk (e.g., multi-dose packaging), or subunit dose. The instructions provided in the kit of the present invention are typically written instructions on a label or package insert (e.g., paper included in the kit), but machine-readable instructions (e.g., instructions carried on a magnetic or optical storage disk) are also acceptable.

[0110] The label or package insert indicates that the composition is used to treat, delay the onset of, and / or alleviate an IgE-associated disorder.Instructions for practicing any of the methods described herein can be provided.

[0111] The kits of the present disclosure are in suitable packaging. Suitable packaging includes, but is not limited to, vials, bottles, jars, flexible packaging (e.g., sealed Mylar or plastic bags), and the like. Packaging used in conjunction with specific devices, such as inhalers, nasal administration devices (e.g., nebulizers), or infusion devices such as miniature pumps, is also contemplated. The kit may have a sterile inlet (e.g., the container may be an intravenous solution bag or a vial with a stopper pierceable by a hypodermic needle). The container may also have a sterile inlet (e.g., the container may be an intravenous solution bag or a vial with a stopper pierceable by a hypodermic needle). At least one active agent in the composition is an anti-Cεmx antibody, such as FB825.

[0112] The kit may optionally provide additional components, such as buffer and interpretative information. Typically, the kit includes a container and a label or one or more package inserts on or associated with the container. In some embodiments, the disclosure provides an article comprising the contents of the above-mentioned kit.

[0113] General Technology

[0114] The practice of the present invention will employ, unless otherwise indicated, conventional techniques of molecular biology (including recombinant techniques), microbiology, cell biology, biochemistry, and immunology, which are within the skill of the art. Such techniques are fully explained in the literature, such as Molecular Cloning: A Laboratory Manual, 2nd Edition (Sambrook et al., 1989) Cold Spring Harbor Press; Oligonucleotide Synthesis (MJ Gait, ed., 1984); Methods in Molecular Biology, Humana Press; Cell Biology: A Laboratory Notebook (JE Cellis, ed., 1998) Academic Press; Animal Cell Culture (RI Freshney, ed., 1987); Introduction to Cell and Tissue Culture (JP Thayer and PE Roberts, 1998) Plenum Press; Cell and Tissue Culture: Laboratory Procedures (A. Doyle, JB Griffiths and DG Newell, eds., 1993-8) J. Wiley and Sons; Methods in Enzymology (Academic Press, Inc.); Handbook of Experimental Immunology (DM Weir and CC Blackwell, eds.); Gene Transfer Vectors for Mammalian Cells (JM Weir and CC Blackwell, eds.); Cells (JM Miller and MP Calos, 1987); Current Protocols in Molecular Biology (FM Ausubel et al., 1987); PCR: The Polymerase Chain Reaction, (Mullis et al., 1994); Current Protocols in Immunology (JE Coligan et al., 1991); Short Protocols in Molecular Biology (Wiley and Sons, 1999); Immunobiology (CA Janeway and P. Travers, 1997); Antibodies (P.Finch, 1997); Antibodies: a practical approach (D. Catty., editor, IRL Press, 1988-1989); Monoclonal antibodies: a practical approach (P. Shepherd and C. Dean, editors, Oxford University Press, 2000); Using antibodies: a laboratory manual (E. Harlow and D. Lane (Cold Spring Harbor Laboratory Press, 1999); The Antibodies (edited by M. Zanetti and JD Capra, Harwood Academic Publishers, 1995).

[0115] Without further elaboration, it is believed that those skilled in the art can utilize the present invention to the greatest extent based on the above description. Therefore, the following specific embodiments should be interpreted as merely illustrative and not limiting the remainder of the present disclosure in any way. All publications cited herein are incorporated by reference for the purpose or subject matter cited herein.

[0116] Example 1: Toxicity study of FB825 in cynomolgus monkeys

[0117] Materials and methods

[0118] Laboratory testing

[0119] Blood and urine samples for hematology, coagulation, serum chemistry (including liver function tests), thyroid function tests, and urinalysis will be collected and analyzed as per routine clinical laboratory testing.

[0120] Clinical laboratory abnormal values ​​are marked as high or low (or normal or abnormal) according to the reference range of each laboratory parameter. Clinically significant is defined as any change in the results that are medically relevant and can lead to a change in medical care (e.g., active observation, diagnostic measures, or therapeutic measures). If a clinically significant change relative to screening is noted, the clinically significant value and clinically significant reason are recorded. The treated cynomolgus monkeys will continue to be monitored and additional assessments will be performed until the value reaches the reference range or the value at screening, or until medical follow-up is no longer necessary.

[0121] Pharmacodynamic evaluation

[0122] Blood samples for determination of total IgE and anti-drug antibodies (ADA) were collected using 3.5-mL blood collection tubes (BD Biosciences, Inc., New York, NY) provided by Vince and Associates Clinical Research. SST TM Serum SeparationTubes) are collected. One sample is collected, with the minimum blood volume for each blood collection tube size as the goal. At least 1.0mL serum is collected at each time point. After obtaining the blood sample, the collection tube is inverted 5 times, and the blood is allowed to coagulate for 30 minutes at ambient temperature (19°C-24°C). The sample is centrifuged at about 2200rpm for 10 minutes at room temperature in a rotary bucket centrifuge. Standard laboratory techniques are used to transfer approximately equal volumes of duplicate serum aliquots (each aliquot is at least 500 μL) to 2 appropriately labeled storage tubes (2-mL polypropylene frozen vials) provided by Vince and Associates Clinical Research.

[0123] Analyze the immunogenicity samples for measuring ADA using a validated ELISA assay.

[0124] Serum aspartate aminotransferase and alanine aminotransferase

[0125] Liver function in FB825-treated cynomolgus monkeys was monitored by measuring the levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT). The activities of these two enzymes were expressed as units per liter (U / L).

[0126] result

[0127] Single-dose study of FB825 in cynomolgus monkeys

[0128] The single-dose toxicity of FB825 was evaluated in non-GLP single-dose toxicity studies conducted in cynomolgus monkeys. In the first study, treatment-related effects were determined in cynomolgus monkeys that were administered a single 10-minute intravenous infusion of FB825. In the second study, treatment-related effects were determined in cynomolgus monkeys that were administered a single subcutaneous injection of FB825.

[0129] In the intravenous injection-dose-range discovery study, male and female cynomolgus monkeys tolerated a single 10-minute intravenous infusion of FB825 well at 30, 100 and 300 mg / kg. There were no treatment-related effects on clinical parameters, food consumption, body weight or mortality. The FB825-related effects were limited to minimal increases in ALT and AST (recovered by day 57) and minimal increases in interleukin-6 and interleukin-10 (at 6 hours and / or 1 hour after administration in animals administered at least 100 mg / kg FB825). Based on these results, the no-observation-adverse-effect level (NOAEL) was considered to be 300 mg / kg.

[0130] In another single-dose toxicology study, FB825 was well tolerated in cynomolgus monkeys via a single subcutaneous injection at 300 mg / kg. No FB825-related clinical signs or effects on body weight or clinical pathology parameters (hematology, coagulation, and clinical chemistry) were observed.

[0131] Repeated-dose study of FB825 in cynomolgus monkeys

[0132] Repeated dose toxicity of FB825 was evaluated in cynomolgus monkeys. Treatment-related effects were determined in cynomolgus monkeys administered a single 10-minute intravenous infusion of FB825 once weekly (4 doses).

[0133] FB825 administered by 10-minute intravenous infusion once weekly was well tolerated by cynomolgus monkeys at 30, 100 and 300 mg / kg (4 doses in total). The following parameters and endpoints were evaluated in this study: clinical signs, body weight, food consumption, ophthalmology, electrocardiogram, clinical pathology parameters (hematology, coagulation and clinical chemistry), bioanalysis and toxicokinetics evaluation, anti-therapeutic antibody analysis, flow cytometry, IgE analysis, thyroid hormone levels, gross autopsy findings, organ weights and histopathological examinations.

[0134] No FB825-related clinical signs or effects on body weight, food consumption, ophthalmological and electrocardiographic examinations, coagulation parameters, and thyroid hormone levels were observed in either sex at doses up to 300 mg / kg. In addition, there were no FB825-related gross findings at doses up to and including 300 mg / kg.

[0135] FB825-related effects were limited to reversible significant increases in ALT at FB825 doses greater than or equal to 100 mg / kg, and reversible mild elevations in AST, partially reversible minimal decreases in albumin levels and corresponding albumin:globulin ratios, and reversible minimal increases in monocyte counts at a FB825 dose of 300 mg / kg. The magnitude of the increases in ALT and AST levels at 300 mg / kg was considered adverse. Target organ effects were observed at levels ≥30 mg / kg and consisted of nonadverse follicular colloid depletion of the thyroid gland and lower thyroid weights. However, the lower thyroid weights were within the range observed in historical control monkeys. Because changes in colloid staining patterns, changes in thyroid follicle size, and vacuolar degeneration have been reported spontaneously in cynomolgus monkeys (Ishida 2000; Hatakeyama 2011), and because there were no test article-related effects on thyroxine and thyroid-stimulating hormone levels, the thyroid findings were considered nonadverse under the conditions of this study.

[0136] Based on the increase in ALT and AST levels at 300 mg / kg, the NOAEL was considered to be 100 mg / kg (C max 5330 μg / mL and AUC (0-168h) 520 mg·hr / mL for men; and C max 5220 μg / mL and AUC (0-168h) is 487 mg·hr / mL for women).

[0137] Example 2: Human clinical studies

[0138] The primary objective of this study was to evaluate the safety and tolerability of single ascending intravenous doses of FB825 in normal healthy subjects. Secondary objectives of this study included determining the PK profile of single ascending intravenous doses of FB825, to explore the effect on total IgE after single ascending intravenous doses of FB825, and to explore the development of anti-FB825 antibodies after single ascending intravenous doses of FB825.

[0139] Study Design

[0140] This is a Phase 1, first-in-human (FIH), randomized, double-blind, placebo-controlled study to evaluate the safety, tolerability, pharmacokinetics, and immunogenicity of single ascending intravenous doses of FB825. Table 1 describes an overview of the event schedule conducted during the FB825 human clinical study. Subjects who met the study inclusion criteria were assigned to the current dose cohort and were randomly assigned to receive FB825 or placebo (vehicle). All doses of FB825 were administered as a 1-hour intravenous infusion.

[0141] Safety data for each dose cohort were reviewed prior to progression to the next highest dose cohort. Dosing of the next highest dose cohort was permitted only after safety was confirmed to be so. Dose levels may be adjusted or repeated as needed. Blinded PK data were reviewed concurrently with blinded safety data.

[0142] The study included 6 groups and a total of approximately 54 normal healthy subjects (7 subjects per group [4 active: 3 placebo] in groups A and B, and 10 subjects per group [7 active: 3 placebo] in groups C, D, E and F). The initial FB825 dose was 0.003 mg / kg IV, with planned increases to 0.03, 0.3, 1.5, 5 and 10 mg / kg IV in subsequent groups. The study consisted of a screening period (days -28 to -2), a check-in (day -1), a treatment / follow-up period (days 1 to 140), and an end-of-study visit (day 140).

[0143] Subjects signed in to the clinic on Day -1 and underwent the sign-in procedure. When procedures overlapped and were scheduled to be performed at the same time point, the order of procedures was vital sign measurements, electrocardiogram, and then pharmacokinetic blood collection.

[0144] Subjects received active drug or placebo study drug on the 1st day. The first two subjects from the 2 lowest dose groups (groups A and B) received placebo or FB825 (that is, both placebo and active study drug need to be indicated). The remaining subjects in these groups were administered 48 hours after the first 2 subjects were administered. All subjects in the 4 highest dose groups were administered at the same time. At least 14 days were separated between the administration of the last subject in the group, and then it was decided to continue to administer to the next group. Before progressing to the second highest dose group, the safety data of each dose group were checked in a blinded manner. Only after confirming that it is safe to do so, administration in the second highest dose group is allowed. Blind PK data and blind safety data are checked simultaneously.

[0145] Blood samples for PK assessment were collected at the following time points on Day 1: 30 minutes before the start of infusion (±5 minutes); 30 minutes after the start of infusion (±2 minutes); 1, 1.25 and 2 hours after the start of infusion (±2 minutes); 4 and 8 hours after the start of infusion (±5 minutes); and 24 and 48 hours after the start of infusion (±10 minutes). In addition, single blood samples were collected on days 5, 14, 29, 85 and 140 after infusion. Blood samples for immunogenicity assessment were collected on Day 1 (30 minutes [±5 minutes] before the start of infusion) and on days 5, 14, 29, 85 and 140.

[0146] Subjects were confined to the clinic from Day -1 until discharge on Day 3 (48 hours post-dose) and returned for outpatient visits on Days 5, 14, 29, 85, and 140. The duration of the study excluding screening was approximately 140 days.

[0147] Patient population

[0148] Healthy male and female subjects aged 18 to 55 years (inclusive) had a body weight greater than or equal to 50 kg and a body mass index of 18.0 to 30.0 kg / m2 (inclusive), and provided written informed consent. A total of 54 subjects were recruited, 41 subjects (75.9%) completed the study, and 13 subjects (24.1%) were stopped. Seven subjects (13.0%) stopped due to subject selection, and included 1 subject each in the 0.3, 1.5, 5 and 10 mg / kg FB825 treatment groups, and included 3 subjects in the placebo treatment group. 6 subjects (11.1%) lost follow-up, and included 1 subject each in the 0.003, 0.3 and 1.5 mg / kg FB825 and placebo treatment groups, and included 2 subjects in the 10 mg / kg FB825 treatment group.

[0149] Table 1. Timeline of events

[0150]

[0151] Abbreviations: ADA, antidrug antibodies; ECG, electrocardiogram; EOS, end of study; FSH, follicle-stimulating hormone; Ig, immunoglobulin.

[0152] FB825 Application

[0153] On day 1, subjects received a single intravenous infusion of FB825 (0.003, 0.03, 0.3, 1.5, 5, or 10 mg / kg) or placebo over approximately 1 hour.

[0154] All doses were administered as a single, approximately 1-hour intravenous infusion on the morning of the scheduled dosing day (Day 1), after fasting for approximately 2 hours. Light meals were allowed 2 hours or more before dosing. Water was allowed at any time except 1 hour before and 1 hour after dosing. The FB825 solution was diluted before administration.

[0155] A single, approximately 1-hour intravenous infusion of FB825 or placebo was administered at 0 hours on the first day. The progress of the next dose level depends on the approval of the safety review team. The first 2 subjects from the 2 lowest dose groups (groups A and B) received placebo or FB825 (i.e., both placebo and active agent need to be represented). The remaining subjects in these groups were administered 48 hours after the first 2 subjects were administered. All subjects in the 4 highest dose groups were administered simultaneously.

[0156] Pharmacokinetic (PK) evaluation

[0157] Blood samples for PK analysis of FB825 were collected from all subjects at the following time points: on day 1, 30 minutes (±5 minutes) before the start of infusion; 30 minutes (±2 minutes) after the start of infusion; 1, 1.25 and 2 hours after the start of infusion (±2 minutes, 30 minutes to 2 hours after administration); 4 and 8 hours after the start of infusion (±5 minutes, 4 to 8 hours after administration); and 24 and 48 hours after the start of infusion (±10 minutes, 24 to 48 hours after administration). In addition, single blood samples were collected at 5, 14 (±1 day), 29 (±2 days), 85 (±3 days) and 140 (±5 days) after infusion.

[0158] The following single-dose PK parameters of FB825 were calculated from the serum concentration data for each subject using a non-compartmental approach:

[0159] AUC 0-t The area under the serum concentration-time curve (AUC) from time 0 to the last quantifiable concentration was calculated using the linear trapezoidal rule

[0160] AUC 0-inf The AUC extrapolated from time 0 to infinity is calculated using the following formula: AUC 0-inf =AUC 0-t +C t / K el , where C t is the last measurable serum concentration, and K el is the terminal elimination rate constant. If the extrapolated area (C t / K el ) is greater than AUC 0-inf 20%, then the AUC 0-inf and its related parameters (CL and V d ) is set to missing.

[0161] %AUC ex For calculating AUC 0-inf Extrapolated area percentage

[0162] C maxMaximum observed serum concentration

[0163] T max Time of maximum observed serum concentration

[0164] K el The terminal elimination rate constant, K el is the magnitude of the linear regression slope of the log concentration versus time curve during the terminal phase. 2 ≥0.80 and 3 points in the terminal stage excluding C max , keep K el .

[0165] t 1 / 2 Terminal half-life (whenever possible), expressed as (ln)2 / K el calculate

[0166] MRT average dwell time, calculated as AUMC / AUC

[0167] CL Apparent clearance, by dose / AUC 0-inf calculate

[0168] V d The apparent volume of distribution is calculated as follows: dose / (AUC 0-inf ×K el )

[0169] Pharmacokinetic blood samples

[0170] Blood was collected using 3.5 mL blood collection tubes (BD SST TM Blood samples were collected for analysis of FB825 using serum separation tubes. One sample was collected, targeting the minimum blood volume for each blood collection tube size. A minimum of 1.0 mL of serum was collected at each time point. After obtaining the blood sample, the collection tube was inverted 5 times and the blood was allowed to coagulate for 30 minutes at ambient temperature (19°C-24°C). The sample was centrifuged at approximately 2200 rpm for 10 minutes at room temperature in a rotary bucket centrifuge. Using standard laboratory techniques, approximately equal volumes of duplicate serum aliquots (at least 500 μL per aliquot) were transferred to 2 appropriately labeled storage tubes (2-mL polypropylene cryovials) provided by Vince and Associates Clinical Research. Labels were affixed to each storage tube and contained the following information:

[0171] Sample type: Human serum

[0172] Assay Type: PK

[0173] Solution: FB825CLCT01

[0174] Subject number: random number

[0175] Time point: Reference plan

[0176] Serum aliquots: 1 or 2

[0177] Within 90 minutes of collection, two aliquot samples were stored vertically at -70°C ± 10°C. Pharmacokinetic serum assays of FB825 were performed using a validated ELISA.

[0178] Pharmacodynamic evaluation

[0179] Blood samples for determination of total IgE and anti-drug antibodies (ADA) were collected on day 1 30 minutes (±5 minutes) before the start of infusion, and on days 5, 14 (±1 day), 29 (±2 days), 85 (±3 days), and 140 (±5 days) after infusion or at early termination.

[0180] Blood was collected using 3.5 mL blood collection tubes (BD SST TM Serum Separation Tubes) are used to collect blood samples for analyzing ADA levels. One sample is collected, with the minimum blood volume of each blood collection tube size as the goal. At least 1.0mL serum is collected at each time point. After obtaining the blood sample, the collection tube is inverted 5 times, and the blood is coagulated for 30 minutes at ambient temperature (19°C-24°C). The sample is centrifuged at room temperature for 10 minutes at about 2200rpm in a rotary bucket centrifuge. Using standard laboratory techniques, two copies of serum aliquots of approximately equal volume (at least 500 μL per aliquot) are transferred to 2 appropriately labeled storage tubes (2-mL polypropylene frozen tubes). Use a validated ELISA to analyze the immunogenicity samples for ADA measurement.

[0181] Security Assessment

[0182] Safety and tolerability were assessed by monitoring and recording AEs, clinical laboratory test results (hematology, coagulation, serum chemistry including liver function tests, thyroid function tests, and urinalysis), vital sign measurements, 12-lead electrocardiogram results, cardiac telemetry data, and physical examination results.

[0183] Clinical laboratory tests

[0184] Blood and urine samples were collected under fasting conditions (fasting for approximately 2 or more hours) at the time points indicated in the event schedule (Table 1) for hematology, coagulation, serum chemistry (including liver function tests), thyroid function tests, urinalysis, and drug screening tests. Clinical laboratory tests (hematology, coagulation, serum chemistry [including liver function tests], and urinalysis) were performed at screening; check-in; on Day 1 before the start of the infusion and at 1 (end of infusion), 8, and 24 hours (±15 minutes) after the start of the infusion; and at a single time point on Days 3 (±15 minutes, continuing up to 48 hours after dosing), 5, 14, 29, 85, and 140.

[0185] Samples were used for clinical laboratory testing including hematology, coagulation, serum chemistry, thyroid function, and urinalysis.

[0186] All female subjects underwent serum pregnancy testing (β-human chorionic gonadotropin) at Screening, Check-in, Day 3 (48 hours after dosing), and End of Study Visit (Day 140). Postmenopausal female subjects underwent serum FSH testing at Screening.

[0187] Hepatitis B surface antigen, antibodies to hepatitis C virus, and antibodies to human immunodeficiency virus (type 1 or 2) were assessed at screening.

[0188] At screening and on day -1, alcohol, amphetamines, barbiturates, benzodiazepines Urine drug screen for cocaine metabolites, cotinine, MDMA, opiates, phencyclidine, propoxyphene, and THC.

[0189] Abnormal clinical laboratory values ​​are marked as high or low (or normal or abnormal) according to the reference range for each laboratory parameter. Clinically significant is defined as any change in the results that are medically relevant and may lead to a change in medical care (e.g., active observation, diagnostic measures, or therapeutic measures). If a clinically significant change relative to screening is noted, the clinically significant value and clinically significant reason are recorded on the AE page of the eCRF. Subjects are continuously monitored with additional assessments until the value reaches the reference range or the value at screening, or until medical follow-up is no longer necessary.

[0190] Vital signs measurement

[0191] Vital sign measurements included systolic and diastolic blood pressure, heart rate, respiratory rate, and oral temperature. Subjects were seated for at least 5 minutes before all measurements were taken, except for the orthostatic assessment. At the time point for the orthostatic assessment, after all measurements were taken for seated subjects, subjects were placed in a supine position for 5 minutes before their blood pressure and heart rate were obtained; subjects were then placed in a standing position for 1 minute before their blood pressure and heart rate were obtained. Vital signs were measured at the time points indicated in the event schedule (Table 1).

[0192] Vital sign measurements (systolic and diastolic blood pressure, heart rate, respiratory rate, and oral temperature) were obtained at screening; at check-in; on Day 1 before the start of infusion, and at 1 (end of infusion), 2 and 4 hours (±15 minutes), and 8 and 24 hours (±30 minutes) after the start of infusion; and on Days 3 (±15 minutes, continuing up to 48 hours after dosing), 5, 14, 29, 85, and 140. Vital sign measurements were taken every 15 minutes (±5 minutes) during the infusion. Orthostatic assessments were performed at check-in; on Day 1 before the start of infusion, and at 2, 4, 8, and 24 hours (±15 minutes) after the start of infusion; and on Day 5.

[0193] Clinically significant is defined as any change in the outcome that is medically relevant and may lead to a change in medical care (e.g., active observation, diagnostic measures, or therapeutic measures). If a clinically significant change relative to screening is noted, the clinically significant value and clinically significant reason are recorded on the AE page of the subject's eCRF. Subjects may be continuously monitored by additional assessments until the value reaches the reference range or the value at screening, or until medical follow-up is no longer necessary.

[0194] 12-lead ECG

[0195] A single 12-lead ECG was obtained at the time points indicated in the event schedule, with the patient in the supine position for at least 5 minutes (Table 1). A 12-lead ECG was obtained at screening; check-in; on Day 1, within 2 hours before the start of the infusion and at 1 (end of infusion), 8, and 24 hours (±15 minutes) after the start of the infusion; and on Days 3 (±15 minutes, continuing up to 48 hours after dosing), 5, 14, and 140.

[0196] The ECG evaluation included comments on whether the trace was normal or abnormal and on the rhythm, presence of arrhythmias or conduction defects, morphology, evidence of myocardial infarction, and abnormalities of the ST segment, T wave, and U wave. In addition, the following intervals were measured and reported: RR interval, PR interval, QRS width, QT interval, and QTcF.

[0197] Clinically significant is defined as any change in the outcome that is medically relevant and may lead to a change in medical care (e.g., active observation, diagnostic measures, or therapeutic measures). If a clinically significant change relative to screening is noted, the clinically significant value and clinically significant reason are recorded on the AE page of the subject's eCRF. Subjects are continuously monitored with additional assessments until any value reaches the reference range or the value at screening, or until medical follow-up is no longer necessary.

[0198] Cardiac Telemetry

[0199] Cardiac telemetry was performed at the time points specified in the event schedule (Table 1). Cardiac telemetry monitoring began on Day 1 (beginning approximately 30 minutes before the start of the infusion) and continued for 4 hours after the end of the infusion.

[0200] Clinically significant is defined as any change in outcome that is medically relevant and may lead to a change in medical care (e.g., active observation, diagnostic measures, or therapeutic measures). If a clinically significant change is noted relative to the initial telemetry findings on Day 1 (approximately 30 minutes prior to the start of the infusion), the clinically significant value and clinically significant reason are recorded on the AE page of the subject's eCRF. Subjects are monitored continuously with additional assessments until medical follow-up is no longer necessary.

[0201] Physical examination

[0202] Complete and abbreviated physical examinations were performed at the time points specified in the event schedule (Table 1). Complete physical examinations (including assessment of skin erythema) were performed at screening and on Days 3, 14, and 140. Abbreviated physical examinations (including assessment of skin erythema) were performed at check-in (Day -1) and on Days 5, 29, and 85. The physical examination included height and weight at screening and weight on other days only.

[0203] A complete physical examination included evaluation of the skin (including any signs of skin erythema), head, ears, eyes, nose, throat, neck, thyroid, lungs, heart, cardiovascular system, abdomen, lymph nodes, and musculoskeletal system / limbs. Interim physical examinations were performed to evaluate for AEs or clinical laboratory abnormalities.

[0204] The brief physical examination includes evaluation of the skin (including any signs of skin erythema), lungs, cardiovascular system, and abdomen (liver, spleen).

[0205] Height and weight were measured, and body mass index was calculated only at Screening. Weight was measured at all other examination time points indicated in the event schedule.

[0206] Dose selection for human studies

[0207] The starting dose in this Phase 1 FIH study was 0.003 mg / kg, which was determined using the NOAEL from a 4-week repeat-dose toxicology study in non-human primates (the closest and most relevant model to humans) and applying appropriate safety factors and from calculating the minimum anticipated biological effect level (MABEL) for FB825 based on in vitro and in vivo pharmacology and toxicokinetic data.

[0208] In a 4-week repeat dose toxicology study in cynomolgus monkeys, the NOAEL of FB825 was considered to be 100 mg / kg. According to the U.S. Food and Drug Administration (FDA) industry guidance: the maximum safe starting dose estimated in initial clinical trials for treatment of adult healthy volunteers (July 2005), the calculated equivalent human dose is 38.8 mg / kg; after applying an appropriate safety factor (i.e., 100 times) to this, the maximum recommended starting dose in human subjects is about 0.39 mg / kg.

[0209] The MABEL for FB825 is estimated to be 0.1 μg / mL. It is estimated that a single intravenous dose of 0.003 mg / kg in human subjects would result in a systemic exposure range of 0.06 μg / mL to 0.09 μg / mL to FB825; therefore, the MABEL should not be exceeded during the study. Taking the higher number of this range, in cynomolgus monkeys administered a single intravenous dose of FB825 (Study 20031008), the expected maximum circulating concentration of FB825 at this starting dose would be higher than the C of FB825 at the dose level of the NOAEL (100 mg / kg / day). max Low about 4.4×10 4 times. In mg / kg, this provides 3.7×10 4 times the safety margin, exceeding and higher than the human equivalent dose of 111.6 mg / kg under NOAEL (300 mg / kg).

[0210] result

[0211] This study evaluated the safety, tolerability, pharmacokinetics and immunogenicity of FB825 via single ascending doses of intravenous injection in a randomized, placebo-controlled, double-blind study of healthy adult subjects. The design and selection of the study population for the planned first-in-human (FIH) clinical phase 1 study were based on the need to obtain initial safety, tolerability, and PK epidemiological results of FB825 for further clinical research. Data were obtained according to the event schedule shown in Table 1.

[0212] FB825 is safely administered to human subjects

[0213] FB825 was infused intravenously in a single 1-hour dose at a dose of 0.003. 0.03, 0.3, 1.5, 5 and 10 mg / kg were safe and well tolerated by healthy subjects in clinical trials. There were no deaths, and no subjects discontinued due to treatment-emergent adverse events (TEAEs). Except for decreased hemoglobin, upper respiratory tract infection, urinary tract infection and gunshot wounds, all TEAEs resolved by the end of the study.

[0214] There were no significant treatment- or dose-related trends in clinical laboratory test results, vital sign measurements, 12-lead ECG results, or physical examination findings.

[0215] Reduction of total IgE by administration of FB825

[0216] Total IgE decreased at all post-dose time points in the 1.5 and 10 mg / kg treatment groups. Total IgE decreased at some time points post-dose in the other doses (0.003, 0.03, 0.3, and 5 mg / kg FB825) and placebo treatment groups, but there was no overall trend ( Figure 1 and Figure 2 ).

[0217] Only 4 subjects (1 each in the 0.03, 0.3, and 5 mg / kg FB825 and placebo treatment groups) had detectable ADA, of which only 1 subject in the 0.03 mg / kg FB825 treatment group had reactive ADA ( Figure 3 ).

[0218] Example 3: Open-label exploratory study evaluating the safety and efficacy of FB825 in adults with atopic dermatitis

[0219] This study was designed to evaluate the changes from baseline in total IgE and allergen-specific IgE in patients with atopic dermatitis after intravenous administration of FB825, and to evaluate the clinical efficacy in these patients. This study also aims to evaluate the safety of FB825 in patients treated thereby, monitor changes in clinical hematology after intravenous administration of FB825, and explore changes from baseline in biomarkers after intravenous administration of FB825, including thymus and activation-regulated chemokine (TARC), Eotaxin-3, thymic stromal lymphopoietin (TSLP), periostin, IL-1a, IL-4, IL-5, IL-13, IL-16, IL-31, and M-CSF.

[0220] FB825 is a humanized monoclonal immunoglobulin G1 (IgG1) that targets the CεmX domain on human B lymphocytes expressing membrane-bound IgE (mIgE). FB825 can block the biological pathway of IgE synthesis, thereby benefiting the treatment of IgE-mediated allergic diseases. FB825 is formulated into an aqueous solution and administered to patients at 5 mg / kg via a 1-hour intravenous infusion.

[0221] Study Procedure

[0222] This is an open-label exploratory study to evaluate the safety and efficacy of FB825 in adults with atopic dermatitis (AD). Approximately 12 human patients with atopic dermatitis (AD) who met the study inclusion criteria were recruited to the study. All eligible patients received 5 mg / kg FB825 by 1 hour intravenous infusion on days 1 and 85 (Table 2). Subjects were hospitalized after receiving FB825 on days 1 and 85 and were discharged the next day for safety observation (at least 12 hours). Patients returned to the study site on days 8, 15, 29, 57, 85, 92, 99, 113, 141, and 169 for safety and efficacy and biomarker evaluation (Table 2).

[0223] In some cases, patients may be asked to make an on-site visit on day 78 for blood sampling. The purpose of blood sampling is to measure total IgE to determine whether the subject needs a second dose of FB825. Subjects whose total IgE changes less than 50% from baseline on day 78 can receive a second dose of FB825 (5 mg / kg). Subjects whose total IgE changes 50% from baseline on day 78 can end the study visit on day 85 and then complete the study. For patients receiving a second dose of FB825, the data collected for the second dose can be analyzed and summarized in the same way as those collected for the first dose. In some cases, patients receive a second dose of FB825 (5 mg / kg) by 1 hour intravenous infusion on day 85. Such patients were hospitalized before receiving FB825 on day 85 and were discharged the next day for safety observation (at least 12 hours). Patients returned to the study site on days 92, 99, 113, 141 and 169 for safety and efficacy evaluation.

[0224] Serum total IgE and antigen-specific IgE were measured at scheduled visits and evaluated to explore changes from baseline following intravenous administration of 5 mg / kg FB825.

[0225] Patients were examined at scheduled visits for clinical efficacy assessment activities, including the pruritus visual analogue scale (VAS), the eczema area and severity index (EASI), the severity score of atopic dermatitis index (SCORAD), the investigator global assessment (IGA) of AD and the body surface area (BSA) involving AD symptoms.

[0226] The PI reviewed safety data including AEs and laboratory tests. The duration of subject participation in the study was approximately 24 weeks.

[0227] Selection of study population

[0228] Male or female subjects with atopic dermatitis were recruited to a single study site. A total of approximately 15 subjects were recruited to achieve at least 12 evaluable subjects.

[0229] Selection criteria

[0230] (i) Main inclusion criteria

[0231] Male or female subjects aged 20 to 65 years (inclusive).

[0232] The subject has a physician-confirmed diagnosis of chronic atopic dermatitis as defined by the Hannifin and Rajka criteria modified by Eichenfeld and supported by a 3-year history of symptoms with positive allergen-specific IgE at the screening visit.

[0233] Eczema Area and Severity Index (EASI) score ≥ 14 at screening and baseline visits.

[0234] Investigator's Global Assessment (IGA) score ≥ 3 (out of 5) at screening and baseline visits.

[0235] AD involving ≥10% body surface area (BSA) at screening and baseline visits.

[0236] History of inadequate response to stable (1 month) topical corticosteroid therapy or calcineurin inhibitors as treatment for AD within 3 months prior to the screening visit. (Topical corticosteroid regimen means moderate to high potency, applied for at least 28 days or the maximum duration recommended by the product prescribing information.)

[0237] • Patients must have been applying a stable dose of an emollient provided for atopic dermatitis twice daily for at least 7 days prior to the baseline visit.

[0238] Female subjects of childbearing potential must use at least two forms of birth control. One must be barrier protection (i.e., condoms or female condoms) and the other must be one of the acceptable birth control methods (i.e., diaphragm, intrauterine contraceptive device, hormonal contraceptive, or abstinence pills) throughout the study. Subjects who are surgically infertile (i.e., hysterectomy, bilateral tubal ligation, or bilateral oophorectomy) or postmenopausal (defined as 12 consecutive months of amenorrhea and recorded follicle-stimulating hormone levels >40mU / mL) will be considered to have no reproductive potential. All female subjects must have a negative serum pregnancy test prior to dosing. Subjects must use the contraceptive methods mentioned above during the study and for 16 weeks or 5 half-lives after the last dose of FB825.

[0239] Subjects weighed ≥40 kg at screening and had a body mass index of 18.0 to 30.0 kg / m 2 (inclusive).

[0240] The subject has a normal 12-lead electrocardiogram (ECG) as determined by the Investigator with normal cardiac conduction parameters of:

[0241] - Heart rate 45 to 100 bpm;

[0242] - Friccria-corrected QT interval (QTcF) ≤450 milliseconds (males) or ≤470 milliseconds (females)

[0243] -QRS interval is less than 120 milliseconds.

[0244] If the investigator determines that the subject is healthy, except for atopic disease, based on the results of clinical laboratory tests performed at screening. If the results are outside the normal reference range, the subject may be included only if the investigator judges that the abnormality or deviation from normal levels is not clinically significant. This determination must be recorded in the subject's original file and initiated by the investigator. This does not apply to laboratory abnormalities listed in the exclusion criteria (using the Microbiology and Infectious Diseases Department criteria).

[0245] ·The subjects were able to provide written informed consent.

[0246] ·The subject agrees to comply with all protocol requirements.

[0247] (ii) Main exclusion criteria

[0248] · Pregnant or breastfeeding female subjects.

[0249] The subject is dieting or not eating enough.

[0250] Subject has a history of cardiac arrhythmia (any clinically relevant).

[0251] The subject has a positive test result for hepatitis B surface antigen, hepatitis C virus antibody, or human immunodeficiency virus antibody at screening.

[0252] The investigator believes that the subject has a history of alcohol or drug abuse that would impair or jeopardize the subject's full participation in the study.

[0253] The subject is subject to judicial supervision or regulation.

[0254] Subjects with clinically relevant, current active or potential gastrointestinal cardiovascular disease, neurological disease, psychiatric disease, metabolic disease, renal disease, hepatic disease, respiratory disease (except uncomplicated allergic rhinitis), inflammatory disease, immunological disease, endocrine disease, diabetes or infectious disease and are not eligible to participate in the study as determined by the investigator.

[0255] Subjects have any previous history of allergic reactions.

[0256] The subject has any medical condition that, in the opinion of the investigator, would impair the study or the subject's health or prevent the subject from meeting or performing the study requirements.

[0257] The subject has received any immunoglobulin products or blood products within three months before dosing.

[0258] Subject has received a biological product:

[0259] - Subject has received any cytodepleting agent, not limited to rituximab, within 6 months prior to dosing or before lymphocyte counts returned to normal, whichever is longer.

[0260] - Subject has received other biologics within 5 half-lives (if known) or 16 weeks (whichever is longer) prior to dosing.

[0261] Subject has one or more of the following laboratory abnormalities at Screening as defined in the Division of Microbiology and Infectious Diseases Adult Toxicity Table, 2007 [Laboratory values ​​may be converted to equivalent standard units. Retesting of abnormal laboratory values ​​that may lead to exclusion is permitted once (without prior approval by the Sponsor). Retesting will be performed during an unscheduled visit (prior to baseline) during the Screening Period]:

[0262] - Aspartate aminotransferase or alanine aminotransferase (>2x upper limit of normal [ULN]) or higher;

[0263] - Total bilirubin ≥1.5x ULN

[0264] - Serum creatinine ≥1.6 x ULN

[0265] - Any other laboratory abnormality higher than or equal to Grade 2 except IgE level, eosinophil count, eosinophil cationic protein (ECP) and the laboratory values ​​mentioned above.

[0266] Laboratory values ​​may be converted to equivalent standard units. Retesting of abnormal laboratory values ​​that may lead to exclusion may be permitted once (without prior approval by the Sponsor). Retesting may be performed during an unscheduled visit (prior to baseline) during the Screening Period.

[0267] The subject has received any approved or unapproved (i.e., investigational) immunotherapy within the past 3 months.

[0268] The subject has used any of the following medications (prescription or over-the-counter):

[0269] -Intranasal corticosteroids (e.g., fluticasone propionate) within 30 days prior to administration.

[0270] -Systemic corticosteroids (e.g., prednisone) within 30 days prior to administration.

[0271] - Leukotriene modifiers (e.g., montelukast) within 30 days prior to dosing.

[0272] -Immunosuppressants (eg, gold salts, methotrexate, azathioprine, cyclosporine) within 30 days prior to administration.

[0273] - Immunomodulatory drugs (such as IFN-γ) within 30 days prior to administration.

[0274] - Anti-IgE within the past 1 year prior to administration of medication (e.g., omalizumab).

[0275] - Allergen immunotherapy within the past 1 year prior to dosing.

[0276] -Oral inhaled corticosteroids (e.g., budesonide) within the past 30 days prior to dosing.

[0277] The subject had received phototherapy within 4 weeks prior to dosing.

[0278] The subject has received a live vaccine within 12 weeks prior to dosing.

[0279] Subjects with a known or suspected history of immunosuppression, including a history of opportunistic infections (e.g., TB), as determined by the investigator.

[0280] · Subjects with a history of malignant disease within 5 years before the screening period.

[0281] High risk of parasitic infection. Risk factors for parasitic disease (living in an endemic area, chronic gastrointestinal symptoms, travel within the past 6 months to an area where geohelminth infection is endemic, and / or chronic immunosuppression) and evidence of parasitic colonization or infection at the time of stool evaluation for ova and parasites. Stool evaluation for ova and parasites should only be performed in patients with risk factors and an eosinophil count greater than twice the upper limit of normal for subjects.

[0282] Study Treatment

[0283] (i) Dosage and intravenous administration of study treatment

[0284] Two doses of 5 mg / kg FB825 were given to subjects with atopic dermatitis. On the morning of Day 1 and Day 85, subjects received FB825 via a 1-hour intravenous infusion. In some cases, fasting for at least 2 hours was required. No water was allowed within 1 hour before and after administration. The amount of FB825 administered can be adjusted according to the subject's weight, and the appropriate amount of the drug is diluted with 250 mL of 0.9% sodium chloride solution. FB825 is administered via the intravenous route within 1 hour with the aid of a programmable volume infusion device. After reconstitution, the final diluted product containing 0.9% sodium chloride must be used as soon as possible and must be used within 8 hours. The reconstituted FB825 can be stored at a temperature of 2°C to 25°C for up to 8 hours before use.

[0285] (ii) Prior, concomitant, and contraindicated medications

[0286] Previous medications and therapies

[0287] The subject's CRF recorded information about previous medications taken by the subject within 30 days before he or she provided informed consent.

[0288] Pretreatment / concomitant medications and procedures

[0289] Any treatment (including nutritional supplements) or procedure administered between the time the ICF is signed and the end of the study visit is considered concomitant and recorded on the CRF. This includes licensed medications that were ongoing at the time of consent.

[0290] The basic AD therapy during the study period is described as follows:

[0291] - All patients applied a moisturizer at least twice daily for at least 7 consecutive days prior to dosing and continued treatment throughout the study. All types of moisturizers were permitted, however patients were not allowed to initiate treatment with prescription moisturizers or moisturizers containing additives during the screening period or during the study.

[0292] - Patients may continue to use a stable dose of prescription moisturizers or moisturizers with additives if started prior to the screening visit. Beginning on Day 1 / Baseline, all patients are to initiate topical corticosteroid (TCS) therapy using a standardized regimen following the following guidelines:

[0293] -Apply medium-potency TCS daily to areas with active lesions. Low-potency TCS should be used in thin skin areas (face, neck, tricuspid and genital areas, skin atrophy, etc.), or in areas where continued treatment with medium-potency TCS is considered unsafe.

[0294] -After lesions are controlled (clear or almost clear), switch from intermediate-acting TCS to moderate-acting TCS and continue daily treatment for 7 days, then stop.

[0295] - If lesions recur, reinstitute treatment with intermediate-acting TCS using the step-down approach described above after lesions have resolved.

[0296] - For lesions that persist or worsen despite daily treatment with intermediate-potency TCS, patients can be treated with high-potency or ultra-potency TCS (rescue) unless high-potency TCS is considered unsafe.

[0297] -Monitor patients for signs of local or systemic TCS toxicity and taper or discontinue treatment as needed.

[0298] - Record the type and amount of topical product used during the study. Determine the amount of TCS used by weighing the tube at each visit (see Study Reference Manual for details).

[0299] -Advise patients to use fluticasone propionate 0.05% cream, mometasone furoate 0.1% cream, or betamethasone valerate 0.06% cream as intermediate-potency TCS, and hydrocortisone 1% ointment for low-potency TCS.

[0300] -If rescue with TCS is required, patients are advised to use fluocinolone acetonide 0.05% cream, desoximetasone 0.25% ointment for high-efficiency TCS, and clobetasol propionate 0.05% ointment for ultra-high-efficiency TCS.

[0301] -Do not use moisturizer and TCS on the same area during the day. On areas not treated with TCS, apply moisturizer twice daily (morning and evening).

[0302] Pre-treatment medications / procedures: medications taken or procedures performed before administration of a drug.

[0303] Concomitant Medication / Procedure: Medication taken or procedure performed after intravenous administration of study medication at the EOS visit.

[0304] Concomitant medications prohibited:

[0305] -Topical tacrolimus and pimecrolimus

[0306] - Systemic corticosteroids, unless the subject is receiving rescue medication.

[0307] -Leukotriene inhibitors

[0308] -Allergen Immunotherapy

[0309] - Systemic treatment for AD with immunosuppressive / immunomodulatory substances (including but not limited to cyclosporine, mycophenolate mofetil, IFN-γ, azathioprine, methotrexate, or biologics)

[0310] -Treatment with live (attenuated) vaccines

[0311] -Traditional Chinese Medicine

[0312] Prohibited accompanying programs:

[0313] -Surgical procedures

[0314] - Concomitant ultraviolet (UV) procedures (phototherapy [NBUVB, UVB, UVA1, or PUVA])

[0315] - Tanning in bed / cabin is not allowed during the study

[0316] - Patients were not permitted to take more than 2 bleach baths per week during study participation.

[0317] (iii) Management of infusion-related or allergic reactions

[0318] Intravenous administration of study drug must be performed under the supervision of trained medical personnel and in facilities available for the management of allergic reactions. If a subject experiences an infusion-related reaction, it must be treated symptomatically with supportive care, further monitoring, and appropriate medical therapy, which may include antihistamines and / or corticosteroids as necessary. The study infusion may be stopped, and the subject will be followed until the end of the study. The infusion volume will be recorded. If a subject experiences typical symptoms of an allergic reaction (e.g., shortness of breath, anaphylactic reaction, urticaria, angioedema), intravenous administration of study drug should be immediately and permanently discontinued.

[0319] Suspected anaphylaxis should be evaluated according to the clinical diagnostic criteria outlined by the National Institute of Allergy and Infectious Diseases provided in Appendix 12-2.

[0320] For these and other conditions, subjects may receive appropriate medical treatment at the discretion of the investigator.

[0321] If an allergic reaction occurs, the patient may be saved by treating the intolerable AD symptoms with contraindicated medications or procedures.

[0322] - Rescue for AD with systemic corticosteroids at a dose of less than 1 mg / kg / day prednisolone and for no more than 3 days may be offered to study subjects after Week 2 if medically necessary (ie, to control intolerable AD symptoms).

[0323] -Perform efficacy and safety assessments (e.g., disease severity scores, safety labs) on patients immediately prior to administration of any rescue therapy.

[0324] Research procedures and evaluation methods

[0325] The following sections describe the study procedures and data to be collected. Subjects were assessed by the same investigator or site personnel whenever possible. The schedule and assessments are provided in Table 2. Baseline characteristics are provided in Table 3.

[0326] (i) end:

[0327] Primary endpoint:

[0328] - Change from baseline in total IgE at Day 85 and Day 169 / End of Study (EOS).

[0329] Change from baseline in allergen-specific IgE at Day 85 and Day 169 / EOS.

[0330] Biomarker endpoints:

[0331] - Change from baseline in total IgE on Days 8, 15, 29, 57, 85, 92, 99, 113, 141, and 169.

[0332] - Changes from baseline in allergen-specific IgE on days 8, 15, 29 and 57, 85, 92, 99, 113, 141, and 169. Changes from baseline in biomarkers including thymus and activation-regulated chemokine (TARC), Eotaxin-3, thymic stromal lymphopoietin (TSLP), periostin, IL-1a, IL-4, IL-5, IL-13, IL-16, IL-31, and M-CSF following intravenous administration of FB825 on days 8, 15, 29, 57, 85, 92, 99, 113, 141, and 169.

[0333] Efficacy endpoints:

[0334] - Change from baseline in pruritus visual analogue scale (VAS) on Days 8, 15, 29, 57, 85, 92, 99, 113, 141, and 169.

[0335] - Change from baseline in the Eczema Area and Severity Index (EASI) on Days 8, 15, 29, 57, 85, 92, 99, 113, 141, and 169.

[0336] - Change from baseline in the Severity Score of Atopic Dermatitis Index (SCORAD) on Days 8, 15, 29, 57, 85, 92, 99, 113, 141, and 169.

[0337] - Change from baseline in the Investigator's Global Assessment (IGA) of atopic dermatitis on Days 8, 15, 29, 57, 85, 92, 99, 113, 141, and 169.

[0338] - Change from baseline in body surface area (BSA) of participants with atopic dermatitis on Days 8, 15, 29, 57, 85, 92, 99, 113, 141, and 169.

[0339] - Changes from baseline in biomarkers including thymus and activation-regulated chemokine (TARC), Eotaxin-3, thymic stromal lymphopoietin (TSLP), periostin, IL-1a, IL-4, IL-5, IL-13, IL-16, IL-31, and M-CSF following intravenous administration of FB825 on days 8, 15, 29, 57, 85, 92, 99, 113, 141, and 169.

[0340] -Safety was assessed by monitoring and recording adverse events (AEs) and serious adverse events (SAEs); physical examination findings and vital sign measurements (systolic and diastolic blood pressure, heart rate, respiratory rate, and temperature); clinical laboratory test results (hematology, coagulation, serum chemistry [including liver function tests, blood glucose levels], and urinalysis); and 12-lead ECG results.

[0341] (ii) Biomarker Assessment

[0342] Blood samples for measurement of total allergen-specific IgE, thymic and activation-regulated chemokine (TARC), Eotaxin-3, thymic stromal lymphopoietin (TSLP), periostin, IL-1a, IL-4, IL-5, IL-13, IL-16, IL-31, and M-CSF were analyzed according to methods described in a separate report.

[0343] Collect blood samples. Actual sample collection times and sampling problems (if any) are recorded on the CRF. Draw approximately 5 mL of venous blood for each sample at the following time points:

[0344] Total immunoglobulin / allergen-specific IgE:

[0345] - Screening period: at any time

[0346] -Days 1 and 85: (within 2 hours before start of infusion)

[0347] - Days 8, 15, 29, 57, 92, 99, 113, 141 and 169 after intravenous administration of FB825 at any time of the day.

[0348] ·TARC, Eotaxin-3, TSLP, Periostin, IL-1a, IL-4, IL-5, IL-13, IL-16, IL-31 and M-CSF:

[0349] - Within 2 hours before the start of the infusion on Days 1 and 85, and at any time on Days 8, 15, 29, 57, 92, 99, 113, 141, and 169.

[0350] (iii) Clinical efficacy evaluation

[0351] Pruritus visual analogue scale (VAS).

[0352] Pruritus was measured using the SCORAD Pruritus Visual Analog Scale (VAS). Patients were asked to assign a numerical score representing the intensity of their symptoms on a scale from 0 to 10, with 0 indicating no symptoms and 10 indicating the most severe symptoms. Subjects were asked to take measurements at screening, on days 1 and 85 before dosing, on days 2 and 86 before discharge, and at any time on days 8, 15, 29, 57, 92, 99, 113, 141, and 169.

[0353] Severity Scoring of Atopic Dermatitis Index (SCORAD)

[0354] SCORAD (index) is a validated scoring system in atopic dermatitis (AD). To measure the extent of AD, a nine-point scale is applied to the before / after images of the patient's inflammatory lesions. The scale can range from 0 to 100. The intensity portion of SCORAD consists of 6 items: erythema, edema / papules, excoriations, lichenification, exudation / crusting, and dryness. Each item can be graded on a scale of 0 (absent) to 3 (severe). Subjective items include daily pruritus and insomnia. The SCORAD index formula is: A / 5+7B / 2+C. In this formula, A is defined as the extent (0–100), B is defined as the intensity (0–18), and C is defined as the subjective symptom (0–20). The maximum score of the SCORAD index is 103. Subjects were asked to have measurements taken at screening, on days 1 and 85 before dosing, on days 2 and 86 before discharge, and at any time on days 8, 15, 29, 57, 92, 99, 113, 141, and 169.

[0355] Eczema Area and Severity Index (EASI)

[0356] The EASI scoring system grades the severity and extent of eczema signs using a defined process. The extent and severity of eczema signs are assessed on four body regions, and the total score is the sum of the four region scores adjusted by a multiplier. The EASI score ranges from 0-72. Subjects are asked to have measurements taken at screening, on days 1 and 85 prior to dosing, on days 2 and 86 prior to discharge, and any time on days 8, 15, 29, 57, 92, 99, 113, 141, and 169.

[0357] Investigator Global Assessment (IGA) of AD

[0358] The IGA allows investigators to assess overall disease severity at a given time point and consists of a 5-point severity scale ranging from clear to very severe disease (0 = clear, 1 = almost clear, 2 = mild disease, 3 = moderate disease, and 4 = severe disease). Subjects were asked to take measurements at screening, on days 1 and 85 prior to dosing, on days 2 and 86 prior to discharge, and any time on days 8, 15, 29, 57, 92, 99, 113, 141, and 169.

[0359] BSA involving AD symptoms

[0360] It will be measured as part A (range) of SCORAD. Subjects will be asked to have the measurement at screening, on days 1 and 85 prior to dosing, on days 2 and 86 prior to discharge, and any time on days 8, 15, 29, 57, 92, 99, 113, 141, and 169.

[0361] (iv) Safety Assessment

[0362] Safety was assessed by monitoring and recording adverse events (AEs) and serious adverse events (SAEs), physical examination findings and vital sign measurements (systolic and diastolic blood pressure, heart rate, respiratory rate, and oral temperature), clinical laboratory test results (hematology, coagulation, serum chemistry [including liver function tests], and urinalysis), and 12-lead ECG results. The total number of patients who experienced an adverse event-SAF throughout the 15-168 day treatment period is provided in Table 4.

[0363] Adverse events

[0364] The Adverse Events section (Section 7) describes adverse events (SAEs), serious adverse events (SAEs), and adverse events of special interest collected during the study.

[0365] Physical examination

[0366] A comprehensive physical examination was performed at the time points specified in the timeline of events.

[0367] A complete physical examination included assessment of the skin (including any signs of skin erythema), head, ears, eyes, nose, throat, neck, thyroid, lungs, heart, cardiovascular system, abdomen, lymph nodes, and musculoskeletal system / limbs. Interim physical examinations were performed at the discretion of the investigator, as necessary, to evaluate AEs or clinical laboratory abnormalities. Height and weight were measured, and body mass index was calculated only at screening. Weight was also measured at all other physical examination time points as indicated in the study event schedule. In indoor clothing (without coat and shoes), weight was recorded in kilograms (kg) to one decimal place, and height (without shoes) was measured in centimeters (cm) without decimal places.

[0368] Vital signs measurement

[0369] Vital sign measurements included systolic and diastolic blood pressure, heart rate, respiratory rate, and body temperature. Subjects sat still for at least 5 minutes before all measurements were taken. Vital signs were measured at the time points indicated in the event schedule.

[0370] When procedures overlap and occur at the same time point, the order of procedures should be vital sign measurement followed by ECG.

[0371] The researcher can determine that any vital sign measurement value is clinically significant or not clinically significant. Clinically significant is defined as any change in the results that have medical relevance and can lead to changes in medical care (such as, active observation, diagnostic measures or therapeutic measures). If a clinically significant change relative to the screening value is noted, the clinically significant value and clinically significant reason are recorded on the AE page of the subject's CRF. The researcher can continue to monitor the subject with additional assessments until the value reaches the reference range or the value when screening, or until the researcher determines that medical follow-up is no longer necessary.

[0372] Clinical laboratory tests

[0373] Clinical laboratory tests were performed at local sites. Blood and urine were collected at the time points indicated in the event schedule under fasting conditions (approximately 2 or more hours).

[0374] The following hematology, coagulation, serum chemistry (including liver and thyroid function tests), and urinalysis evaluations were performed:

[0375] -Hematology: Hematocrit (Hct), hemoglobin (Hb), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), mean corpuscular volume (MCV), platelet count, red blood cell (RBC) count, and WBC and differential counts (absolute values ​​and percentages).

[0376] -Coagulation: International Normalized Ratio (INR), Partial Thromboplastin Time (PTT) and -Prothrombin Time (PT).

[0377] - Serum chemistries: ALT, albumin, alkaline phosphatase, amylase, anion gap, AST, bicarbonate, bilirubin (total and direct), blood urea nitrogen (BUN), calcium, carbon dioxide, chloride, cholesterol (total, HDL, and calculated LDL), creatine phosphokinase, creatinine, gamma-glutamyl transferase (γ-GT), globulin, glucose, lactate dehydrogenase (LDH), lipase, magnesium, phosphorus, potassium, sodium, total protein, triglycerides, troponin I or T, and uric acid.

[0378] -Thyroid function: free T4 and TSH

[0379] -Urinalysis: appearance, bilirubin, color, glucose, ketones, leukocyte esterase, microscopy (if dipstick is positive; include bacteria, casts, crystals, epithelial cells, erythrocytes, and leukocytes), nitrites, occult blood, pH, protein, specific gravity, turbidity, and urobilinogen

[0380] A serum pregnancy test (beta-human chorionic gonadotropin) will be performed on all female subjects of reproductive potential at Screening and the result will be confirmed prior to dosing. A urine pregnancy test will be performed on Days 8, 15, 29, 57, 85, 92, 99, 113, 141, and 169. For postmenopausal female subjects, a serum follicle-stimulating hormone test will be performed at Screening.

[0381] Hepatitis B surface antigen, hepatitis C virus antibodies, and human immunodeficiency virus antibodies will be assessed at screening.

[0382] According to the reference range of each laboratory parameter, abnormal clinical laboratory values ​​are marked as high or low (or normal or abnormal). The researcher can determine whether any abnormally high or low result is clinically significant or not clinically significant. Clinically significant is defined as any change in the result that has medical relevance and can cause medical care (such as, active observation, diagnostic measures or therapeutic measures) to change. If a clinically significant change relative to the screening value is noted, the clinically significant value and clinically significant reason can be recorded on the AE page of the CRF. The researcher can continue to monitor the subject with additional assessments until the value reaches the reference range or the value at the time of screening, or until the researcher determines that medical follow-up is no longer necessary.

[0383] Clinically significant laboratory values ​​for individual subjects are listed. A summary of the number and percentage of subjects with clinically significant laboratory values ​​at any time is presented.

[0384] Electrocardiogram

[0385] At the time points indicated in the timeline of events, a single 12-lead ECG is obtained after the patient has been in the supine position for at least 5 minutes. The ECG evaluation includes comments on whether the trace is normal or abnormal, the rhythm, the presence of arrhythmias or conduction defects, the morphology, any evidence of myocardial infarction, or abnormalities of the ST segment, T wave, and U wave. In addition, measurements of the following intervals are measured and reported: RR interval, PR interval, QRS width, QT interval, and QTcF.

[0386] Adverse Events

[0387] The total number of patients who experienced an adverse event-SAF during the entire 15-168 day treatment period is provided in Table 4.

[0388] (i) definition

[0389] Adverse events (AEs)

[0390] An AE is defined as any untoward medical occurrence associated with the use of a drug in humans, whether or not considered drug-related. Subjects will be instructed to contact the investigator at any time during the study if any symptoms occur.

[0391] A TEAE was defined as any event that was not present before exposure to study drug or any pre-existing event that worsened in intensity or frequency after exposure.

[0392] An adverse reaction is any AE attributed to a drug. An adverse reaction is a subset of all suspected adverse reactions for which it is reasonable to conclude that the drug caused the event.

[0393] A suspected adverse reaction is any AE for which there is a reasonable possibility that the study drug caused the AE. For the purposes of investigational new drug safety reporting, "reasonable possibility" means there is evidence of a causal relationship between the study drug and the AE. A suspected adverse reaction means that there is less certainty about the causal relationship than an adverse reaction, which means any AE caused by the study drug.

[0394] An AE or suspected adverse reaction is considered "unexpected" if it is not listed in the IB or is not at the specificity or severity observed with the test study drug; or, if an IB is not required or provided, is inconsistent with the risk information described elsewhere in the general study plan or current application. For example, according to this definition, if the IB refers only to elevated liver enzymes or hepatitis, then hepatic necrosis is unexpected (due to higher severity). Similarly, if the IB lists only cerebrovascular accidents, cerebral thromboembolism and cerebral vasculitis would be unexpected (due to higher specificity). "Unexpected" as used in this definition also refers to AEs or suspected adverse reactions mentioned in the IB that occur with a class of drugs or are expected from the pharmacological properties of the drug, but are not specifically mentioned as occurring with the specific drug being studied.

[0395] Serious adverse events (SAEs)

[0396] An AE or suspected adverse reaction was considered a SAE if, in the opinion of the investigator or sponsor, it resulted in any of the following outcomes:

[0397] -die

[0398] - Life-threatening AEs

[0399] - Inpatient hospitalization or extension of existing hospitalization

[0400] - Permanent or severe disability or severe impairment of the ability to carry out normal life functions

[0401] - Congenital anomalies or birth defects

[0402] Important medical events that would not result in death, life-threatening conditions, or hospitalization may be considered serious when, based on appropriate medical judgment, they may endanger the subject and may require medical or surgical intervention to prevent one of the outcomes listed in this definition. Examples of such medical events include allergic bronchospasm requiring intensive treatment in an emergency department or at home, blood disorders or seizures that do not result in inpatient hospitalization, or the development of drug dependence or drug abuse.

[0403] An AE or suspected adverse reaction is considered “life-threatening” if, in the opinion of the investigator or sponsor, its occurrence places the subject at immediate risk of death. It does not include AEs or suspected adverse reactions that could result in death if they occurred in a more severe form.

[0404] Adverse events of special concern

[0405] Adverse events of particular interest include the following:

[0406] - Any subject experienced a treatment-emergent AE (TEAE) of anaphylaxis

[0407] - Any subject experienced a SAE (Section 7.1.2)

[0408] - Any subject who experiences at least 30 minutes of persistent QT prolongation (>500 milliseconds or change from baseline ≥60 milliseconds) or ischemic changes (on repeat ECG), or persistent symptomatic arrhythmias

[0409] - Hypersensitivity, including anaphylactic reactions, thyroid abnormalities, skin erythema, thrombocytopenia, anemia, hemolysis, neutropenia, hepatotoxicity, and nephrotoxicity.

[0410] - The following laboratory parameters were encountered in any single subject:

[0411] √ALT or AST ≥ 5 × ULN

[0412] √Bilirubin ≥ 2 × ULN

[0413] √ Platelet count level 1≤99.999*109 / L

[0414] √Hemoglobin ≤ 105 g / L

[0415] √ Absolute neutrophil count level 1≤1.5*109 / L

[0416] √ Blood urea nitrogen or creatinine increased to >2×ULN

[0417] (ii) Induced adverse events

[0418] The investigator was responsible for ensuring that all AEs and SAEs were documented in the CRF and reported to Fountain. Adverse events were assessed from the time of the screening period until completion of all study procedures and withdrawal from the study.

[0419] At each study visit or assessment, participants were asked a standard question to elicit any medically relevant changes in their health. They were also asked if they had been hospitalized, had an accident, or taken any new medications or had any changes to their concomitant medication regimen (prescription and over-the-counter).

[0420] In addition to subject observations, AEs were recorded from any data collected on the AE page of the CRF (eg, laboratory values, physical examination findings, and ECG changes) or other documentation related to subject safety.

[0421] Severity Assessment The severity (or intensity) of the AE determined using the Common Terminology Criteria for Adverse Events (CTCAE) was used to grade the severity of infusion reactions.

[0422] - Grade 1 Mild: transient or mild discomfort (<48 hours); no medical intervention / therapy required.

[0423] - Grade 2 Moderate: Mild to moderate limitation in activity - may need some assistance; no or minimal medical intervention / therapy required.

[0424] -Grade 3 Severe: Marked limitation of activity, usually requiring some assistance; medical intervention / treatment, possible hospitalization, is required.

[0425] - Grade 4 Life-threatening: Extreme limitation of activity, requiring substantial assistance; requiring substantial medical intervention / therapy, possible hospitalization or hospice care.

[0426] Changes in the severity of AEs should be recorded so that an assessment of the duration of events at each level of intensity can be made. AEs characterized as intermittent require the occurrence and duration of each episode to be recorded.

[0427] 7.3.2 Causality Assessment

[0428] The investigator's assessment of the relationship of the AE to the study drug was part of the documentation process but was not a factor in determining what was or was not reported in the study.

[0429] The investigator assessed the causality of all AEs and SAEs (i.e., whether there was a reasonable possibility that the study drug caused the event). The relationship was characterized using the following categories:

[0430] - Unrelated: This relationship indicates that there is no association between the study drug and the reported event.

[0431] - Probable: The relationship is based on evidence that there is a causal relationship between the study drug and the AE, i.e., there is a reasonable possibility that the drug caused the event. The event follows a reasonable time sequence from intravenous administration of the drug or follows a known pattern of response to the study drug, but it is also possible that it was caused by other factors.

[0432] - Probable: This relationship indicates that there is a plausible temporal sequence of events related to the intravenous administration of the drug, and an association of the events with the study drug appears likely based on the known pharmacological actions of the drug, known or previously reported adverse reactions to the drug or drugs of this class, or the investigator's clinical experience.

[0433] - Clear: This relationship demonstrates that there is a clear causal relationship between intravenous administration of the drug and the AE, and other circumstances (intercurrent illness, progression / expression of a disease state, or concurrent drug reactions) do not appear to explain the event.

[0434] Statistical analysis

[0435] (i) Overall Statistics

[0436] Details of all statistical analyses are described in the statistical analysis plan. All data collected are presented in the data listings. Data for subjects excluded from the analysis population are presented in the data listings but are not included in the calculation of summary statistics.

[0437] For categorical variables, frequencies and percentages are given. Continuous variables were summarized using descriptive statistics (number of subjects, mean, median, SD, minimum and maximum).

[0438] Baseline demographics and background variables were summarized by dose and overall for all subjects. The number of subjects participating in the study and the number and percentage of subjects who completed the study were presented. The frequency and percentage of subjects who withdrew or discontinued the study were also summarized, as well as the reasons for withdrawal or discontinuation. The analysis of this study was demonstrated by descriptive statistics for each period and each group. No significant test was applied.

[0439] A statistical analysis plan (SAP) is written to address the statistical analysis in detail. A clinical database lock may occur after the last patient completes the study and all data have been reconciled (i.e., "cleaned").

[0440] (ii) Sample size calculation

[0441] The study is planned to enroll approximately 12 evaluable subjects in total. The sample size for this study was based on clinical and practical considerations rather than on formal statistical power calculations.

[0442] (iii) Analysis Set

[0443] The Full Analysis Set (FAS) is defined as subjects who received at least 1 dose of study drug. All efficacy evaluations will be performed in the FAS population.

[0444] The safety population is defined as subjects who received at least 1 dose of study drug. All safety evaluations will be performed in the safety population.

[0445] (iv) Statistical analysis

[0446] Biomarker analysis

[0447] Concentrations and changes from baseline of total and allergen-specific IgE and biomarkers listed as endpoints were summarized by visit and presented graphically. Changes from baseline in total and allergen-specific IgE were summarized by baseline total IgE concentration (serum IgE>1500 IU / mL or serum IgE≤1500 IU / mL) and FB825 dose administered (1 dose or 2 doses).

[0448] In subjects who received the second dose of FB825, data collected for the second dose were analyzed and summarized in the same manner as data collected for the first dose.

[0449] Clinical efficacy analysis

[0450] Evaluation indices were assessed by PI or combined PI during each visit for each subject. Clinical endpoints were analyzed by visit time point and FB825 dose administered (1 dose or 2 doses) using descriptive statistics (mean values ​​for EASI, SCORAD, IGA, VAS, and BSA; SD, CV, number of subjects).

[0451] The change in average index relative to scheduled access time is presented graphically,

[0452] In subjects who received the second dose of FB825, data collected for the second dose were analyzed and summarized in the same manner as data collected for the first dose.

[0453] Safety analysis

[0454] Adverse events were coded by preferred term and system organ class using the latest version of the Medical Dictionary for Regulatory Activities and summarized by treatment, dose level, and overall. Adverse events were also summarized by severity, relationship to study drug, SAEs leading to study drug discontinuation, and AEs.

[0455] Actual values ​​and changes from baseline for clinical laboratory test results, vital sign measurements, and 12-lead ECG results were summarized by treatment and dose at each time point using descriptive statistics (number of subjects, mean, SD, median, minimum, and maximum). Moving tables were generated for clinical laboratory test results. Clinical laboratory data, vital sign measurements, 12-lead ECG results, and physical examination findings were presented in data tables.

[0456] (v) Handling of lost data

[0457] For descriptive statistics, concentrations below the limit of quantification (BLQ) will be considered as zero. The mean BLQ concentration will be expressed as BLQ, and the SD and CV will be reported as not applicable. Missing concentrations will be excluded from the calculations.

[0458] Last observation carried forward (LOCF) was used to handle missing data. No imputation was used for safety data.

[0459] (vi) Data quality assurance

[0460] Monitor all aspects of the study to ensure compliance with applicable government regulations, current International Conference on Harmonization (ICH) harmonized tripartite guideline E6(R1): Good Clinical Practice and current standard operating procedures. An internal quality review of the data and additional review by clinical supervisors may be required.

[0461] Table 2. Timeline of events and assessments

[0462]

[0463] Table 3. Baseline characteristics

[0464]

[0465] Table 4. General summary of the number of patients experiencing adverse events during the treatment period - SAF, 15-168 days

[0466]

[0467] result

[0468] This is an open-label exploratory study to evaluate the safety and efficacy of FB825 in adults with atopic dermatitis (AD). Twelve eligible human patients with AD were enrolled in the study and received 5 mg / kg FB825 via a 1-hour intravenous infusion on Days 1 and 85. Patients were scheduled to return to the study site for safety and efficacy evaluations according to the event schedule presented in Table 2.

[0469] FB825 is safely administered to human subjects with AD

[0470] On Day 1 and Day 85, FB825 was administered to patients via intravenous infusion at 5 mg / kg over 1 hour. There were no deaths, and no subjects were discontinued due to treatment-emergent adverse events (TEAEs) or treatment-emergent serious adverse events (TE SAEs). Six subjects experienced TEAEs, three of which were possibly drug-related. Five subjects developed upper respiratory tract infections, and two subjects were infected with herpes virus infections. Only one subject was reported to have one of the following: conjunctivitis, rhinorrhea, asthma exacerbation, fever, cough, or prolonged QTC. (Table 4)

[0471] There were no significant treatment- or dose-related trends in clinical laboratory test results, vital sign measurements, 12-lead ECG results, or physical examination findings.

[0472] effect

[0473] The efficacy of FB825 was determined by recording the change from baseline in the Eczema Area and Severity Index (EASI), Investigator Global Assessment (IGA), Severity Score of Atopic Dermatitis Index (SCORAD), and Pruritus Visual Analog Scale (VAS) at any time on Days 1 and 85 before dosing, Days 2 and 86 before discharge, and on Days 8, 15, 29, 57, 92, 99, 113, 141, and 169. Baseline demographic and background variables are summarized in Table 3 for all subjects by dose and overall.

[0474] (i) Compared with baseline, FB825 reduced EASI

[0475] From day 1 to about day 55 after FB825 administration, subjects showed a gradual decrease in EASI (about 65% decrease). EASI scores began to rise after day 55 until the second FB825 administration on day 85, but were still about 40% lower than baseline. On day 113, subjects showed a further decrease in EASI to about 70% decrease, and maintained this level until the end of the study (EOS). Figure 5 ).

[0476] (ii) Compared with baseline, FB825 reduced IGA

[0477] From day 1 to day 55 after FB825 administration, subjects showed a gradual decrease in IGA (by about 30%). IGA scores began to rise after day 55 until the second FB825 administration on day 85, but were still about 15% lower than baseline. On day 113, subjects showed further IGA decreases to about 45% decrease (at the lowest point), and maintained a 35% decrease compared to baseline until the end of the study (EOS). Figure 6 ).

[0478] (iii) Compared with baseline, FB825 reduced SCORAD

[0479] From day 1 to about day 55 after FB825 administration, subjects showed a gradual decrease in SCORAD (about 45% decrease). SCORAD scores began to rise after day 55 until the second FB825 administration on day 85, but were still about 30% lower than baseline. On day 113, subjects showed further SCORAD decreases to about 55% decrease and maintained this level until the end of the study (EOS). Figure 7 ).

[0480] (iv) Compared with baseline, FB825 reduced VAS

[0481] From day 1 to about day 15 after FB825 administration, subjects showed a gradual decrease in VAS (a decrease of about 45%). VAS scores began to rise after day 15 and maintained about a 25% decrease compared to baseline until the second FB825 administration on day 85. On day 113, subjects showed a further decrease in VAS to about a 55% decrease and maintained this level until the end of the study (EOS). Figure 8 ).

[0482] Other Implementations

[0483] All features disclosed in this specification may be combined in any combination. Each feature disclosed in this specification may be replaced by an alternative feature for the same, equivalent or similar purpose. Therefore, unless otherwise expressly stated, each feature disclosed is only an example of a series of equivalent or similar features.

[0484] From the above description, those skilled in the art can easily determine the basic characteristics of the present invention, and without departing from its spirit and scope, various changes and modifications can be made to the present invention to adapt it to various uses and conditions. Therefore, other embodiments are also within the claims. Sequence Listing <110> Fountain Biopharma Inc. <120> Treatment of IgE-mediated allergic diseases <130> F0720.70004WO00 <140> Not yet assigned <141> at the same time <150> US 62 / 579,416 <151> 2017-10-31 <160> 7 <170> PatentIn Version 3.5 <210> 1 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> Synthetic peptides <400> 1 Gly Leu Ala Gly Gly Ser Ala Gln Ser Gln Arg Ala Pro Asp Arg Val 1 5 10 15 Leu <210> 2 <211> 117 <212> PRT <213> Artificial Sequence <220> <223> Synthetic peptides <400> 2 Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu 1 5 10 15 Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Tyr Ser Ile Thr Ser Asp 20 25 30 Tyr Ala Trp Asn Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp 35 40 45 Ile Gly Ser Ile Ser Tyr Ser Gly Ile Thr Gly Tyr Asn Pro Ser Leu 50 55 60 Lys Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe Ser 65 70 75 80 Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Met Gly Tyr Asp Gly Leu Ala Tyr Trp Gly Gln Gly Thr Leu 100 105 110 Val Thr Val Ser Ser 115 <210> 3 <211> 113 <212> PRT <213> Artificial Sequence <220> <223> Synthetic polypeptide <400> 3 Asp Ile Val Met Thr Gln Thr Pro Leu Ser Leu Ser Val Thr Pro Gly 1 5 10 15 Gln Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Ile Val His Ser 20 25 30 Asn Gly Asn Thr Tyr Leu Glu Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 40 45 Pro Gln Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr Leu Lys Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Phe Gln Gly 85 90 95 Ser His Val Pro Pro Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105 110 Arg <210> 4 <211> 117 <212> PRT <213> Artificial Sequence <220> <223> Synthetic polypeptide <400> 4 Asp Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gln 1 5 10 15 Ser Leu Ser Leu Thr Cys Thr Val Thr Gly Tyr Ser Ile Thr Ser Asp 20 25 30 Tyr Ala Trp Asn Trp Ile Arg Gln Phe Pro Gly Asn Lys Leu Glu Trp 35 40 45 Met Gly Ser Ile Ser Tyr Ser Gly Ile Thr Gly Tyr Asn Pro Ser Leu 50 55 60 Lys Ser Arg Ile Ser Val Thr Arg Asp Thr Ser Lys Asn Gln Phe Phe 65 70 75 80 Leu Gln Leu Asn Ser Val Thr Thr Glu Asp Thr Ala Thr Tyr Tyr Cys 85 90 95 Ala Arg Met Gly Tyr Asp Gly Leu Ala Tyr Trp Gly His Gly Thr Thr 100 105 110 Val Thr Val Ser Ala 115 <210> 5 <211> 113 <212> PRT <213> Artificial Sequence <220> <223> Synthetic polypeptide <400> 5 Asp Val Leu Met Thr Gln Thr Pro Leu Ser Leu Pro Val Ser Leu Gly 1 5 10 15 Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Ile Val His Ser 20 25 30 Asn Gly Asn Thr Tyr Leu Glu Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 40 45 Pro Lys Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr Phe Lys Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Tyr Cys Phe Gln Gly 85 90 95 Ser His Val Pro Pro Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 110 Arg <210> 6 <211> 52 <212> PRT <213> Homo sapiens <400> 6 Gly Leu Ala Gly Gly Ser Ala Gln Ser Gln Arg Ala Pro Asp Arg Val 1 5 10 15 Leu Cys His Ser Gly Gln Gln Gln Gly Leu Pro Arg Ala Ala Gly Gly 20 25 30 Ser Val Pro His Pro Arg Cys His Cys Gly Ala Gly Arg Ala Asp Trp 35 40 45 Pro Gly Pro Pro 50 <210> 7 <211> 12 <212> PRT <213> Artificial Sequence <220> <223> Synthetic polypeptide <400> 7 Gly Leu Ala Gly Gly Ser Ala Gln Ser Gln Arg Ala 1 5 10

Claims

1. Use of an antibody that binds to the Cεmx domain of membrane-bound IgE in the preparation of a medicament for treating a disorder associated with immunoglobulin E (IgE), the treatment comprising: (i) administering a first dose of the drug to a human subject in need thereof; and (ii) administering a second dose of the drug to the human subject; wherein the second dose is administered at least 8 weeks and at most 6 months after the first dose; wherein the first dose, the second dose, or both are 1 mg / kg to 10 mg / kg; The antibody comprises a heavy chain variable region (V H ), the heavy chain variable region includes a heavy chain complementary determining region (CDR) 1, a heavy chain CDR2 and a heavy chain CDR3, wherein the heavy chain CDR1, the heavy chain CDR2 and the heavy chain CDR3 are shown in the amino acid sequences of residues 26-36, 51-66 and 98-106 of SEQ ID NO: 2, respectively; and The antibody comprises a light chain variable region (V L ), the light chain variable region includes a light chain CDR1, a light chain CDR2 and a light chain CDR3, wherein the light chain CDR1, the light chain CDR2 and the light chain CDR3 are respectively as shown in the amino acid sequences of residues 24-39, 55-61 and 94-102 of SEQ ID NO:3; and Wherein the condition is atopic dermatitis.

2. Use of an antibody that binds to the Cεmx domain of membrane-bound IgE in the preparation of a medicament for treating atopic dermatitis, the treatment comprising administering a first dose of the medicament to a human subject in need thereof; wherein the first dose is 1 mg / kg to 10 mg / kg; The antibody comprises a heavy chain variable region (V H ), the heavy chain variable region includes a heavy chain complementary determining region (CDR) 1, a heavy chain CDR2 and a heavy chain CDR3, wherein the heavy chain CDR1, the heavy chain CDR2 and the heavy chain CDR3 are shown in the amino acid sequences of residues 26-36, 51-66 and 98-106 of SEQ ID NO: 2, respectively; and The antibody comprises a light chain variable region (V L ), the light chain variable region includes a light chain CDR1, a light chain CDR2 and a light chain CDR3, and wherein the light chain CDR1, the light chain CDR2 and the light chain CDR3 are respectively as shown in the amino acid sequences of residues 24-39, 55-61 and 94-102 of SEQ ID NO:

3.

3. The use of any one of claims 1 to 2, wherein the antibody comprises V H , the V H The amino acid sequence is as shown in SEQ ID NO: 2, and the antibody comprises V L , the V L The amino acid sequence is as shown in SEQ ID NO:

3.

4. The method according to any one of claims 1 to 2, wherein the antibody is a human antibody or a humanized antibody.

5. The use according to any one of claims 1 to 2, wherein the antibody is a full-length antibody or an antigen-binding fragment thereof.

6. The use according to any one of claims 1 to 2, wherein the antibody is formulated into a pharmaceutical composition comprising the antibody, a buffer, a salt and a non-ionic surfactant.

7. The use according to claim 6, wherein the pharmaceutical composition is an aqueous solution with a pH of 5 to 8.

8. The method of claim 7, wherein the buffer is a histidine buffer, the salt is sodium chloride and / or the nonionic surfactant is polysorbate 80.

9. The use according to claim 8, wherein the antibody in the pharmaceutical composition is 10 mg / ml to 30 mg / ml, the concentration of the histidine buffer is 10-30 mM, the concentration of the sodium chloride is 120-160 mM, and the concentration of the polysorbate 80 is 0.01-0.03%.

10. An aqueous formulation comprising an antibody that binds to the Cεmx domain of membrane-bound IgE at a concentration of 10 mg / ml to 30 mg / ml, a buffer comprising amino acids at a concentration of 10-30 mM, a salt at a concentration of 120-160 mM, and a nonionic surfactant at a concentration of 0.01-0.03%, wherein the pH of the aqueous formulation is 5-8; The antibody comprises a heavy chain variable region (V H ), the heavy chain variable region includes a heavy chain complementary determining region (CDR) 1, a heavy chain CDR2 and a heavy chain CDR3, wherein the heavy chain CDR1, the heavy chain CDR2 and the heavy chain CDR3 are shown in the amino acid sequences of residues 26-36, 51-66 and 98-106 of SEQ ID NO: 2, respectively; and The antibody comprises a light chain variable region (V L ), the light chain variable region includes a light chain CDR1, a light chain CDR2 and a light chain CDR3, wherein the light chain CDR1, the light chain CDR2 and the light chain CDR3 are respectively as shown in the amino acid sequences of residues 24-39, 55-61 and 94-102 of SEQ ID NO:

3.

11. The aqueous formulation of claim 10, wherein the antibody comprises V H and V L , the V H is as shown in the amino acid sequence of SEQ ID NO: 2, and said V L The amino acid sequence is as shown in SEQ ID NO:

3.

12. An aqueous formulation as claimed in claim 10 or claim 11, wherein the antibody is an IgG molecule.

13. An aqueous formulation as claimed in claim 10 or claim 11, wherein the amino acid-containing buffer is a histidine buffer, the salt is sodium chloride, and / or the non-ionic surfactant is polysorbate 80.

14. The aqueous formulation of claim 13, wherein the concentration of the antibody is 20 mg / ml, the concentration of the histidine buffer is 20 mM, the concentration of the sodium chloride is 140 mM, and the concentration of the polysorbate 80 is 0.02%, and wherein the pH of the aqueous formulation is 6.5.

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