CN120603605A - Methods for improving bone growth by administering IL-4R antagonists - Google Patents

Abstract

Methods for improving bone growth in a subject are provided. In one aspect, the method comprises administering one or more doses of an interleukin-4 receptor (IL-4R) antagonist, such as an anti-IL-4R antibody or antigen-binding fragment thereof, to a subject having a defect in bone growth.

Description

Methods for improving bone growth by administering an IL-4R antagonist

Cross Reference to Related Applications

The present application was filed on month 22 of 2023 as PCT international patent application requiring priority and rights of U.S. provisional patent application nos. 63/384,816, 63/480,717, and 63/498,946, respectively, filed on month 20 of 2023 and 4, respectively, as filed on month 22 of 2023, 11, and 23, respectively, the contents of which are incorporated herein by reference.

Reference to sequence Listing XML

The application contains a sequence table that has been submitted electronically in XML format. Sequence Listing XML is incorporated herein by reference. The XML file created at 2023, 11, 17 was named 40848_0118wou1_sl. XML and was 267,776 bytes in size.

Technical Field

The present disclosure relates to the use of interleukin-4 receptor (IL-4R) antagonists for improving bone growth.

Background

Children with Atopic Dermatitis (AD) have a risk of low Bone Mineral Density (BMD), which is associated with reduced bone mass, increased prevalence of osteoporosis and fracture risk (Wu et al ANN TRANSL MED,2021,9:40.doi:10.21037/atm-20-4708; lowe et al, J ALLERGY CLIN Immunol 2020, 145:563-571). Factors such as nutrient limitation, vitamin D deficiency, dyssomnia and corticosteroid use result in lower levels of bone alkaline phosphatase (BALP) (markers of bone mineralization) compared to healthy children, as seen in moderate to severe AD children (Silverberg, PEDIATR ALLERGY Immunol,2015, 26:54-61).

The main determinant of life-long risk of Bone fracture and osteoporosis is the size of the peak Bone mass reached during pre-puberty (Diemar et al, bone,2021,146:115879.Doi:10.1016/j. Bone.2021.115879). Low BALP and BMD in moderate to severe AD children may lead to reduced bone mass and a higher prevalence of osteoporosis.

Disclosure of Invention

In one aspect, a method for improving bone growth is provided. In some embodiments, the method comprises:

selecting a subject having a defect in bone growth, wherein the subject is a pediatric or adolescent subject less than 18 years old, and

One or more doses of an interleukin-4 receptor (IL-4R) antagonist are administered to a subject.

In some embodiments, the IL-4R antagonist is an anti-IL-4R antibody or antigen-binding fragment thereof, e.g., comprising one or more of the CDR, HCVR, and/or LCVR sequences shown in Table 1. In some embodiments, the IL-4R antagonist is an anti-IL-4R antibody or antigen binding fragment thereof comprising three HCDRs (HCDR 1, HCDR2, and HCDR 3) and three LCDRs (LCDR 1, LCDR2, and LCDR 3), wherein HCDR1 comprises the amino acid sequence of SEQ ID NO:3, HCDR2 comprises the amino acid sequence of SEQ ID NO:4, HCDR3 comprises the amino acid sequence of SEQ ID NO:5, LCDR1 comprises the amino acid sequence of SEQ ID NO:6, LCDR2 comprises the amino acid sequence LGS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 8.

In some embodiments, the subject has Atopic Dermatitis (AD). In some embodiments, the subject has moderate to severe or severe AD.

In some embodiments, the subject is a pediatric subject less than 12 years of age. In some embodiments, the subject is 6 to 11 years old. In some embodiments, the subject is 6 months to 5 years old.

In some embodiments, the subject is a teenager aged 12 to 17 years old.

In some embodiments, the subject has combined asthma.

In some embodiments, the selecting step comprises selecting a subject that exhibits a level of a bone turnover marker below a threshold, wherein the bone turnover marker is bone specific alkaline phosphatase, carboxy-terminal cross-linked telopeptide of type I collagen (β -CTX), type I procollagen N-terminal propeptide (PINP), insulin-like growth factor 1 (IGF-1), or osteocalcin. In some embodiments, the threshold is an average level of bone transition markers for a population of healthy subjects of the same age as the pediatric or adolescent subject selected.

In some embodiments, the bone transition marker is a bone-specific alkaline phosphatase.

In some embodiments, the IL-4R antagonist is administered at a dose of about 50mg to about 600 mg. In some embodiments, the IL-4R antagonist is administered at a frequency of once per week (QW), once per two weeks (Q2W), once per three weeks (Q3W), or once per four weeks (Q4W). In some embodiments, the IL-4R antagonist is administered at an initial dose of 100-600mg followed by one or more subsequent doses of 50-300mg, wherein each subsequent dose is administered one week to four weeks after the immediately preceding dose.

In some embodiments, the IL-4R antagonist is administered at an initial dose of 200mg followed by one or more subsequent doses of 200 mg.

In some embodiments, the IL-4R antagonist is administered at an initial dose of 300mg followed by one or more subsequent doses of 300 mg.

In some embodiments, the IL-4R antagonist is administered at an initial dose of 400mg followed by one or more subsequent doses of 200 mg.

In some embodiments, the IL-4R antagonist is administered at an initial dose of 600mg followed by one or more subsequent doses of 300 mg.

In some embodiments, the subject is a pediatric subject of 6 to 11 years old or a adolescent of 12 to 17 years old, and wherein the subject has a baseline body weight of ≡60kg, wherein the IL-4R antagonist is administered subcutaneously at an initial dose of 600mg followed by one or more subsequent doses of 300mg q2 w.

In some embodiments, the subject is a adolescent with a baseline weight of <60kg, and the IL-4R antagonist is administered subcutaneously at an initial dose of 400mg followed by one or more subsequent doses of 200mg q2 w.

In some embodiments, the subject is a pediatric subject aged 6 to 11 years and having a baseline body weight of ≡30kg to <60kg, wherein the IL-4R antagonist is administered subcutaneously at an initial dose of 400mg followed by one or more subsequent doses of 200mg q2 w.

In some embodiments, the subject is a pediatric subject aged 6 to 11 years and having a baseline body weight of ≡15kg to <30kg, wherein the IL-4R antagonist is administered subcutaneously at an initial dose of 600mg followed by one or more subsequent doses of 300mg q4 w.

In some embodiments, the subject is a pediatric subject aged 6 to 11 years and having a baseline body weight of ≡15kg to <60kg, wherein the IL-4R antagonist is administered subcutaneously at an initial dose of 300mg on day 1, followed by 300mg on day 15, followed by one or more subsequent doses of 300mg q4w beginning four weeks after administration on day 15.

In some embodiments, the subject is a pediatric subject aged 6 months to 5 years and having a baseline body weight of ≡15kg to <30kg, wherein the IL-4R antagonist is administered subcutaneously at a dose of 300mg q4 w.

In some embodiments, the subject is a pediatric subject aged 6 months to 5 years and having a baseline body weight of ≡5kg to <15kg, wherein the IL-4R antagonist is administered subcutaneously at a dose of 200mg q4 w.

In some embodiments, the IL-4R antagonist is administered for at least 16 weeks.

In some embodiments, the IL-4R antagonist is administered in combination with a topical AD drug. In some embodiments, the local AD drug is TCS.

In some embodiments, treatment with an IL-4R antagonist results in increased bone growth in the subject as measured by an increase in a bone turnover marker selected from the group consisting of bone-specific alkaline phosphatase, β -CTX, PINP, IGF-1, and osteocalcin.

In some embodiments, an anti-IL-4R antibody or antigen-binding fragment thereof comprises a Heavy Chain Variable Region (HCVR) comprising the amino acid sequence of SEQ ID NO. 1 and comprises a Light Chain Variable Region (LCVR) comprising the amino acid sequence of SEQ ID NO. 2. In some embodiments, the anti-IL-4R antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO. 9 and a light chain comprising the amino acid sequence of SEQ ID NO. 10. In some embodiments, the IL-4R antagonist is Dupilumab Li Youshan.

In some embodiments, the IL-4R antagonist is contained in a container selected from the group consisting of a glass vial, a syringe, a prefilled syringe, a pen delivery device, and an auto-injector. In some embodiments, the IL-4R antagonist is contained in a pre-filled syringe. In some embodiments, the prefilled syringe is a single dose prefilled syringe. In some embodiments, the IL-4R antagonist is contained in a pen delivery device. In some embodiments, the IL-4R antagonist is contained in an auto-injector.

In another aspect, a pharmaceutical composition for improving bone growth is provided. In some embodiments, the pharmaceutical composition comprises an interleukin-4 receptor (IL-4R) antagonist. In some embodiments, the IL-4R antagonist is an anti-IL-4R antibody or antigen-binding fragment thereof, e.g., comprising one or more of the CDR, HCVR, and/or LCVR sequences shown in Table 1. In some embodiments, the IL-4R antagonist is an anti-IL-4R antibody or antigen binding fragment thereof comprising three HCDRs (HCDR 1, HCDR2, and HCDR 3) and three LCDRs (LCDR 1, LCDR2, and LCDR 3), wherein HCDR1 comprises the amino acid sequence of SEQ ID NO:3, HCDR2 comprises the amino acid sequence of SEQ ID NO:4, HCDR3 comprises the amino acid sequence of SEQ ID NO:5, LCDR1 comprises the amino acid sequence of SEQ ID NO:6, LCDR2 comprises the amino acid sequence LGS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 8. In some embodiments, the pharmaceutical composition is used to improve bone growth in a pediatric or adolescent subject (e.g., a pediatric or adolescent subject suffering from atopic dermatitis).

In another aspect, provided herein are interleukin-4 receptor (IL-4R) antagonists for use in the preparation of a medicament for improving bone growth. In some embodiments, the IL-4R antagonist is an anti-IL-4R antibody or antigen-binding fragment thereof, e.g., comprising one or more of the CDR, HCVR, and/or LCVR sequences shown in Table 1. In some embodiments, the IL-4R antagonist is an anti-IL-4R antibody or antigen binding fragment thereof comprising three HCDRs (HCDR 1, HCDR2, and HCDR 3) and three LCDRs (LCDR 1, LCDR2, and LCDR 3), wherein HCDR1 comprises the amino acid sequence of SEQ ID NO:3, HCDR2 comprises the amino acid sequence of SEQ ID NO:4, HCDR3 comprises the amino acid sequence of SEQ ID NO:5, LCDR1 comprises the amino acid sequence of SEQ ID NO:6, LCDR2 comprises the amino acid sequence LGS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 8. In some embodiments, the medicament is for improving bone growth in a pediatric or adolescent subject (e.g., a pediatric or adolescent subject suffering from atopic dermatitis).

Other embodiments will be apparent from reading the detailed description that follows.

Drawings

FIG. 1 shows the geometric mean of bone alkaline phosphatase (BALP) (mcg/L) relative to baseline at visit for patients treated with placebo + Topical Corticosteroid (TCS), dumb Li Youshan against 300mg Q4W+TCS, or dumb Li Youshan against 100mg or 200mg Q2W+TCS in a 16 week maternal study (R668-AD-1652; "LIBERTY AD PEDS") or a subsequent open label extension study (R668-AD-1434; "LIBERTY AD PED-OLE"). The visits at weeks 8, 12 and 16 were from the maternal study, and the visit at week 52 was from the open label extension study. Patients treated with placebo + TCS during the maternal study were converted to either degree of common Li Youshan anti 100mg or 200mg q2w or 300mg q4w for open label extension studies. ns, is not obvious, and SE, standard error.

FIG. 2 shows osteocalcin levels (ng/mL) in patients treated with placebo + TCS or Dupulin Li Youshan antibody (100/200 mg Q2W or 300mg Q4W) +TCS at weeks 8, 12, 16 or 52 of treatment. The connecting lines represent data from the same subject. The box plot shows the median value (middle horizontal line) and the quartile range (lower and upper limits of the box) which correspond to the values at the top of the plot.

FIG. 3 shows the level of type I procollagen N-terminal pro peptide (PINP) in patients treated with placebo +TCS or Dupulin Li Youshan anti (100/200 mg Q2W or 300mg Q4W) +TCS at weeks 8, 12, 16 or 52 of the treatment. The connecting lines represent data from the same subject. The box plot shows the median value (middle horizontal line) and the quartile range (lower and upper limits of the box) which correspond to the values at the top of the plot.

FIG. 4 shows insulin-like growth factor 1 (IGF-1) levels (ng/mL) in patients treated with placebo + TCS or Dupulin Li Youshan anti (100/200 mg Q2W or 300mg Q4W) +TCS at weeks 8, 12, 16 or 52 of treatment. The connecting lines represent data from the same subject. The box plot shows the median value (middle horizontal line) and the quartile range (lower and upper limits of the box) which correspond to the values at the top of the plot.

FIG. 5 shows the level of carboxy-terminal cross-terminal peptide (. Beta. -CTX) of type I collagen in patients treated with placebo + TCS or Dupulin Li Youshan anti (100/200 mg Q2W or 300mg Q4W) +TCS at weeks 8, 12, 16 or 52 of treatment. The connecting lines represent data from the same subject. The box plot shows the median value (middle horizontal line) and the quartile range (lower and upper limits of the box) which correspond to the values at the top of the plot.

Figures 6A and 6B show BALP geometric mean values over time for female (6A) and male (6B) patients in the 6-11 year old treatment group. ^a The diagonal lines represent BALP reference intervals for either females or males. ^b After 16 weeks, these patients received positive degree of common Li Youshan anti-treatment at the time of enrollment in the LIBERTY AD PED-OLE test. The visits at weeks 8, 12 and 16 were from the LIBERTY AD PEDS test, and the visit at week 52 was from the LIBERTY AD PED-OLE test. All P <0.05, P <0.01, P <0.001, P <0.0001 compared to the corresponding placebo + TCS. ns, is not obvious, and SE, standard error.

Figure 7 shows BALP geometric mean values for female (upper panel) and male (lower panel) patients in the 6-11 year old treatment group at visit. The visits at weeks 8, 12 and 16 were from the LIBERTY AD PEDS test, and the visit at week 52 was from the LIBERTY AD PED-OLE test. The numbers under the gender and treatment regimen labels represent the median of the group and the range from lower quartile to upper quartile. ^a After 16 weeks, these patients received positive degree of common Li Youshan anti-treatment at the time of enrollment in the LIBERTY AD PED-OLE test. All P <0.05, P <0.01, P <0.001, P <0.0001, compared to the corresponding baseline.

Detailed Description

Definition of the definition

Before describing the present invention, it is to be understood that this invention is not limited to particular methodology and experimental conditions described, as such methods and conditions may vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting, since the scope of the present invention will be limited only by the appended claims.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

As used herein, the term "about" when used in reference to a specifically recited value means that the value may vary by no more than 1% from the recited value. For example, as used herein, the expression "about 100" includes 99 and 101 and all values therebetween (e.g., 99.1, 99.2, 99.3, 99.4, etc.).

As used herein, the term "treating" or the like means temporarily or permanently alleviating symptoms, eliminating the cause of symptoms, or preventing or slowing the appearance of symptoms of the disorder or condition.

As used herein, the term "subject in need thereof" refers to a human or non-human animal having a defect in bone growth. In some embodiments, "defect in bone growth" refers to, for example, a decrease in bone mineral density levels and/or a decrease in levels of biomarkers of bone formation or bone mineralization as compared to a healthy subject or population of subjects. In some embodiments, the term "subject in need thereof" refers to pediatric patients of less than 12 years of age, such as patients from 6 months to 5 years of age or patients from 6 years to 11 years of age. In some embodiments, the term "subject in need thereof" refers to adolescent patients that are ≡12 years of age and <18 years of age. The terms "subject" and "patient" are used interchangeably herein.

As used herein, "atopic dermatitis" or "AD" means an inflammatory skin disease characterized by intense itching (e.g., severe itching) and squamous and dry eczematous lesions. The term "atopic dermatitis" includes, but is not limited to, AD caused by or associated with epidermal barrier dysfunction, allergy (e.g., allergy to certain foods, pollen, mold, dust mites, animals, etc.), radiation exposure, and/or asthma. The present disclosure includes methods of treating moderate to severe or severe AD patients. As used herein, "moderate to severe AD" is characterized by intense itching, extensive skin lesions, which are often complicated by persistent bacterial, viral or fungal infections. Moderate to severe AD also includes chronic AD in patients. In many cases, chronic lesions include thickened plaques of the skin, lichenification and fibrous papules. In general, patients affected by moderate to severe AD also have more than 20% of their body skin affected, or 10% of their skin area affected, in addition to those involving eyes, hands and body folds. Moderate to severe AD is also thought to be present in patients in need of frequent treatment with topical corticosteroids. Patients may also be referred to as suffering from moderate to severe AD when they are resistant or refractory to treatment with topical corticosteroids or calcineurin inhibitors. As used herein, "severe AD" is characterized by the presence of extensive skin lesions, persistent itching, or physical or emotional disabling diseases that significantly impair the quality of life of the patient. In some cases, patients with severe AD also exhibit one or more symptoms such as abrasion, extensive skin thickening, bleeding, exudation and/or skin cracking, and altered pigmentation. In some embodiments, severe AD is refractory to treatment with local therapies, such as local corticosteroids, calcineurin inhibitors, or clenbuterol (crisaborole).

As used herein, the term "TCS" includes class I, class II, class III and class IV topical corticosteroids. Corticosteroids are classified as weak (group I), moderately effective (group II) and effective (group III) and very effective (group IV) based on their activity compared to hydrocortisone (hydrocortisone) according to the anatomical treatment classification system of the world health organization. Group IV TCS (very potent) was 600 times more potent than hydrocortisone and included clobetasol propionate (clobetasol propionate) and halcinonide (halcinonide). Group III TCS (effective) is 50 to 100 times that of hydrocortisone and includes, but is not limited to, betamethasone valerate (betamethasone valerate), betamethasone dipropionate (betamethasone dipropionate), diflurocortone valerate (diflucortolone valerate), hydrocortisone-17-butyrate, mometasone furoate (mometasone furoate) and methylprednisolone acetate (methylprednisolone aceponate). Group II TCS (moderately potent; also interchangeably referred to herein as "intermediate potent") is 2-to 25-fold that of hydrocortisone and includes, but is not limited to, clobetasone butyrate (clobetasone butyrate) and triamcinolone acetonide (triamcinolone acetonide). Group I TCS (mild; also interchangeably referred to herein as "low potency") includes hydrocortisone.

Although any methods and materials similar or equivalent to those described herein can be used in the practice of the present disclosure, typical methods and materials are now described. All publications mentioned herein are incorporated by reference in their entirety.

Therapeutic method

In one aspect, a method for improving bone growth in a subject is provided. In some embodiments, the subject has a defect in bone growth, such as a defect in bone formation or bone metabolism. In some embodiments, the methods comprise administering one or more doses of an interleukin-4 receptor (IL-4R) antagonist, such as an anti-IL-4 ra antibody or antigen-binding fragment thereof as disclosed herein, to a subject.

In some embodiments, the subject is a pediatric subject or adolescent subject less than 18 years old. In some embodiments, the subject is aged from ≡6 months to <18 years. In some embodiments, the subject is aged from ≡6 to <18 years. In some embodiments, the subject is aged from ≡12 to <18 years. In some embodiments, the subject is aged from ≡6 to <12 years old. In some embodiments, the subject is aged from ≡6 months to <12 years. In some embodiments, the subject is aged from ≡6 months to <6 years.

In some embodiments, the subject is a pediatric or adolescent subject having a weight of <60kg at baseline. In some embodiments, the subject is a pediatric or adolescent subject having a weight of <30kg at baseline. In some embodiments, the subject weighs ∈5kg and <30kg at baseline. In some embodiments, the subject weighs ∈5kg and <15kg at baseline. In some embodiments, the subject weighs ∈15kg and <30kg at baseline.

In some embodiments, the subject has an atopic disease. In some embodiments, the subject has AD (e.g., moderate to severe AD or severe AD). In some embodiments, the subject is diagnosed with chronic atopic dermatitis at least 6 months (e.g., at least 9 months or at least 1 year) before the initiation of treatment. In some embodiments, the subject has moderate to severe or severe AD that is inadequate to respond to, or for which topical therapy (e.g., TCS with or without a local calcineurin inhibitor (TCI)) is undesirable (e.g., due to adverse side effects or safety risks). In some embodiments, the subject has moderate to severe or severe AD and is a candidate for systemic therapy.

In some embodiments, the subject has AD (e.g., moderate to severe AD or severe AD) and has one or more concomitant allergic conditions (i.e., does not include AD). In some embodiments, the subject has a concurrent atopic or allergic disorder selected from the group consisting of allergic rhinitis, asthma, food allergy, non-food allergy, allergic conjunctivitis, urticaria, chronic sinusitis, nasal polyps, and eosinophilic esophagitis.

In some embodiments, the subject has a defect in bone growth. In some embodiments, the subject has abnormal bone metabolism relative to a healthy control or population of healthy control subjects. In some embodiments, the subject has reduced bone formation relative to a healthy control or population of healthy control subjects. In some embodiments, the subject has a reduced bone mineral density relative to a healthy control or population of healthy control subjects. In some embodiments, the subject is at risk of a bone fracture. In some embodiments, the subject has a medical history of a bone fracture. In some embodiments, the subject has reduced bone mass. In some embodiments, the subject has osteoporosis (e.g., idiopathic juvenile osteoporosis or secondary osteoporosis). In some embodiments, the subject with a defect in bone growth has a history of treatment with a local therapy (e.g., a local corticosteroid, calcineurin inhibitor, or clenbuterol).

In some embodiments, the subject with a defect in bone growth is selected based on the level of a bone-specific marker (e.g., a bone formation marker or a bone turnover marker). In some embodiments, the marker is bone-specific alkaline phosphatase, carboxy-terminal cross-linked terminal peptide of type I collagen (CTX-1), type I procollagen N-terminal propeptide (PINP), insulin-like growth factor 1 (IGF-1), or osteocalcin. In some embodiments, the subject is selected based on the level of a marker that exhibits a threshold value (e.g., a bone transition marker).

In some embodiments, the subject with a defect in bone growth is selected based on the subject's Bone Mineral Density (BMD) (e.g., Z-score calculated for one or more bone sites). In some embodiments, the subject is selected based on having a BMD Z score below a threshold. In some embodiments, a subject is identified as having a defect in bone growth if the subject has a BMD Z score of +.2.0, e.g., as measured for lumbar vertebra, femur, hip, or another skeletal site.

In some embodiments, a "threshold" of a parameter or marker (e.g., bone transition marker or BMD Z score) as disclosed herein is determined by reference to a healthy subject population that is the same as or within the age range of a selected pediatric or adolescent subject. For a given parameter or marker, one skilled in the art can determine a threshold for a particular age or range of ages based on knowledge in the art of the level of the parameter or marker in the general population. For example, methods for calculating average BMD Z scores for different age ranges in pediatric and adolescent subjects with AD or healthy control subjects are disclosed in Leung et al, hong Kong Med J,2017,23:470-479; pedreira et al, pediatr Dermatol,2007,24:613-620; penterrich et al, J Pediatr Endocrinol Metab,2018,31:247-260; silverberg et al, J ALLERGY CLIN Immunol,2013,132:1132-1138; silverberg et al, PEDIATR ALLERGY Immunol,2015;26:54-61; and Wu et al, ANN TRANSL MED,2021, 9:40.doi:10.21037/atm-20-4708. Methods for calculating the average levels of Bone formation/Bone turnover markers, including Bone alkaline phosphatase, osteocalcin, PINP, IGF-1 and beta-CTX, are disclosed in Diemar et al, bone,2021,146:115879, penterrich et al, J Pediatr Endocrinol Metab,2018,31:247-260, silverberg et al, PEDIATR ALLERGY Immunol,2015, 26:54-61, and Tobiume et al, J Clin Endocrinol Metab,1997, 82:2056-2061. In some embodiments, the threshold is a lower value for a 95% reference interval for a Bone transition marker established for pediatric or adolescent patients, e.g., as shown in table 3 of Diemar et al, bone,2021,146:115879, or as provided by Mayo Clinic Laboratories Pediatric Catalog (pediatric.

In some embodiments, the subject is selected based on a level of bone alkaline phosphatase exhibiting a threshold value, e.g., a lower value for the 95% reference interval of bone alkaline phosphatase established for pediatric or adolescent patient populations. In some embodiments, a subject is selected if the subject's serum bone alkaline phosphatase level is <70 μg/L, <65 μg/L, <60 μg/L, or <55 μg/L.

In some embodiments, if the subject:

age 8-9 years and serum bone alkaline phosphatase level <53.4 μg/L (for female subjects) or <46.2 μg/L (for male subjects), or

Age 10-11 years and serum bone alkaline phosphatase level <50.6 μg/L (for female subjects) or <52.7 μg/L (for male subjects), or

Age 12-13 years and serum bone alkaline phosphatase level <54.6 μg/L (for female subjects) or <49.5 μg/L (for male subjects), or

Age 14-15 years and serum bone alkaline phosphatase level <14.2 μg/L (for female subjects) or <30.1 μg/L (for male subjects), or

Age 16-17 years, and serum bone alkaline phosphatase level <12.3 μg/L (for female subjects) or <25.7 μg/L (for male subjects), then the subject is selected.

In some embodiments, the subject is selected based on a level of osteocalcin exhibiting a threshold value below, for example, a lower value of the 95% reference interval of osteocalcin established for a pediatric or adolescent patient population. In some embodiments, if the subject:

age 8-9 years and serum osteocalcin level <68.5 μg/L (for female subjects) or <54.1 μg/L (for male subjects), or

Age 10-11 years and serum osteocalcin level <72.2 μg/L (for female subjects) or <55.8 μg/L (for male subjects), or

Age 12-13 years and serum osteocalcin level <82.9 μg/L (for female subjects) or <58.7 μg/L (for male subjects), or

Age 14-15 years and serum osteocalcin level <22.2 μg/L (for female subjects) or <54.1 μg/L (for male subjects), or

Age 16-17 years, and serum osteocalcin level <18.8 μg/L (for female subjects) or <61.5 μg/L (for male subjects), then the subject is selected.

In some embodiments, the subject is selected based on a level of PINP that exhibits a threshold value below, for example, a lower value of the 95% reference interval of PINP established for pediatric or adolescent patient populations. In some embodiments, if the subject:

Age 8-9 years and serum PINP levels <415 μg/L (for female subjects) or <381 μg/L (for male subjects), or

Age 10-11 years, and serum PINP levels <352 μg/L (for female subjects) or <298 μg/L (for male subjects), or

Age 12-13 years and serum PINP levels <387 μg/L (for female subjects) or <168 μg/L (for male subjects), or

Age 14-15 years and serum PINP levels <65 μg/L (for female subjects) or <219 μg/L (for male subjects), or

Age 16-17 years, and serum PINP levels <55 μg/L (for female subjects) or <166 μg/L (for male subjects), then the subject is selected.

In some embodiments, the subject is selected based on a level of β -CTX exhibiting a threshold value below, for example, a lower value of the 95% reference interval of β -CTX established for pediatric or adolescent patient populations. In some embodiments, if the subject:

age 8-9 years, and serum beta-CTX level <1030ng/L (for female subjects) or <1080ng/L (for male subjects), or

Age 10-11 years, and serum beta-CTX level <1103ng/L (for female subjects) or <1140ng/L (for male subjects), or

Age 12-13 years and serum beta-CTX level <960ng/L (for female subjects) or <1100ng/L (for male subjects), or

Age 14-15 years and serum beta-CTX level <330ng/L (for female subjects) or <1000ng/L (for male subjects), or

Age 16-17 years, and serum β -CTX level <290ng/L (for female subjects) or <1060ng/L (for male subjects), then the subject is selected.

In some embodiments, the subject is selected based on exhibiting a level of IGF-1 below a threshold, e.g., a lower value for the 95% reference interval of IGF-1 established for a pediatric or adolescent patient population. In some embodiments, if the subject:

Age <1 year old and serum IGF-1 level <14ng/mL (for female subjects) or <18ng/mL (for male subjects), or

Age 1 year old and serum IGF-1 level <23ng/mL (for female subjects) or <14ng/mL (for male subjects), or

Age 2 years and serum IGF-1 level <28ng/mL (for female subjects) or <16ng/mL (for male subjects), or

Age 3 years, and serum IGF-1 level <31ng/mL (for female subjects) or <22ng/mL (for male subjects), or

Age 4 years and serum IGF-1 level <33ng/mL (for female subjects) or <30ng/mL (for male subjects), or

Age 5 years, and serum IGF-1 level <36ng/mL (for female subjects) or <39ng/mL (for male subjects), or

Age 6 years, and serum IGF-1 level <39ng/mL (for female subjects) or <47ng/mL (for male subjects), or

Age 7 years, and serum IGF-1 level <44ng/mL (for female subjects) or <54ng/mL (for male subjects), or

Age 8 years, and serum IGF-1 level <51ng/mL (for female subjects) or <61ng/mL (for male subjects), or

Age 9 years, and serum IGF-1 level <61ng/mL (for female subjects) or <67ng/mL (for male subjects), or

Age 10 years, and serum IGF-1 level <73ng/mL (for female subjects) or <73ng/mL (for male subjects), or

Age 11 years, and serum IGF-1 level <88ng/mL (for female subjects) or <79ng/mL (for male subjects), or

Age 12 years, and serum IGF-1 level <104ng/mL (for female subjects) or <84ng/mL (for male subjects), or

Age 13 years, and serum IGF-1 level <120ng/mL (for female subjects) or <90ng/mL (for male subjects), or

Age 14 years, and serum IGF-1 level <136ng/mL (for female subjects) or <95ng/mL (for male subjects), or

Age 15 years, and serum IGF-1 level <147ng/mL (for female subjects) or <99ng/mL (for male subjects), or

Age 16 years, and serum IGF-1 level <153ng/mL (for female subjects) or <104ng/mL (for male subjects), or

Age 17 years, and serum IGF-1 level <149ng/mL (for female subjects) or <107ng/mL (for male subjects), or

Age 16-17 years, and serum IGF-1 level <55 μg/L (for female subjects) or <166 μg/L (for male subjects), then the subject is selected.

In some embodiments, treatment with an IL-4R antagonist improves bone growth, improves or normalizes bone turnover, or reduces the severity of a bone defect (e.g., reduces the occurrence or severity of a bone fracture, or reduces the severity of reduced bone mass or osteoporosis).

In some embodiments, treatment with an IL-4R antagonist improves one or more bone-related parameters in a subject. Examples of "bone related parameters" include, but are not limited to, bone formation or bone turnover markers such as bone specific alkaline phosphatase, beta-CTX, PINP, IGF-1, and osteocalcin, and bone mass density, for example, as measured by dual energy X-ray absorptiometry (DEXA). An "improvement in a bone-related parameter" refers to an improvement (e.g., an increase or normalization) in one or more parameters relative to baseline. The term "baseline" as used with respect to bone related parameters means the value of the bone related parameter of the subject prior to or at the beginning of administration of the pharmaceutical composition as disclosed herein.

To determine whether a bone related parameter has "improved," the parameter is quantified at one or more time points after baseline and administration of the pharmaceutical composition of the present disclosure. For example, bone-related parameters may be measured on day 1, day 2, day 3, day 4, day 5, day 6, day 7, day 8, day 9, day 10, day 11, day 12, day 14, day 15, day 22, day 25, day 29, day 36, day 43, day 50, day 57, day 64, day 71, day 85, or on week 1, week 2, week 3, week 4, week 5, week 6, week 7, week 8, week 9, week 10, week 11, week 12, week 13, week 14, week 15, week 16, week 17, week 18, week 19, week 20, week 21, longer, week 22, week 23, week 24, or ending after initial treatment with a pharmaceutical composition of the disclosure. The difference between the parameter value at a particular point in time after initiation of treatment and the parameter value at baseline is used to determine whether there is an "improvement" (e.g., a decrease) in the bone-related parameter.

In some embodiments, treatment with an IL-4R antagonist according to the methods of the present disclosure results in increased bone growth in a subject, as measured by an increase in a bone turnover marker selected from the group consisting of bone-specific alkaline phosphatase, β -CTX, PINP, IGF-1, and osteocalcin. In some embodiments, treatment with the IL-4R antagonist results in an increase in the level of the marker relative to baseline at week 4, week 8, week 12, week 16, week 24, week 30, week 36, week 48, or week 52 after administration of the first dose of the IL-4R antagonist. In some embodiments, treatment with the IL-4R antagonist results in an increase in the level of the marker by at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, or at least 90% relative to baseline at week 4, week 8, week 12, week 16, week 24, week 30, week 36, week 48, or week 52 after administration of the first dose of the IL-4R antagonist.

In some embodiments, treatment with an IL-4R antagonist according to the methods of the present disclosure results in an increase in bone mass in a subject, as measured by Bone Mineral Density (BMD) Z score. In some embodiments, treatment with the IL-4R antagonist results in an improvement or normalization of the subject's BMD Z score relative to baseline at week 4, week 8, week 12, week 16, week 24, week 30, week 36, week 48, or week 52 after administration of the first dose of the IL-4R antagonist. In some embodiments, treatment with the IL-4R antagonist results in an increase in BMD Z score of at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, or at least 90% relative to baseline at week 4, week 8, week 12, week 16, week 24, week 30, week 36, week 48, or week 52 after administration of the first dose of the IL-4R antagonist.

Interleukin-4 receptor antagonists

In some embodiments, the methods of the present disclosure comprise administering an interleukin-4 receptor (IL-4R) antagonist or a pharmaceutical composition comprising an IL-4R antagonist to a subject in need thereof (e.g., a subject with a defect in bone growth). As used herein, an "IL-4R antagonist" (also referred to herein as an "IL-4R inhibitor", "IL-4R blocker" or "L-4 ra antagonist") is any agent that binds or interacts with IL-4 ra or an IL-4R ligand and inhibits or reduces the normal biological signaling function of type 1 and/or type 2 IL-4 receptors. Human IL-4Rα has the amino acid sequence of SEQ ID NO. 11. Type 1 IL-4 receptors are dimeric receptors comprising an IL-4Rα chain and a yc chain. Type 2 IL-4 receptors are dimeric receptors comprising an IL-4Rα chain and an IL-13Rα 1 chain. Type 1 IL-4 receptors interact with and are stimulated by IL-4, while type 2 IL-4 receptors interact with and are stimulated by IL-4 and IL-13. Thus, IL-4R antagonists useful in the methods of the present disclosure may act by blocking IL-4 mediated signaling, IL-13 mediated signaling, or both IL-4 and IL-13 mediated signaling. Thus, the IL-4R antagonists of the present disclosure may prevent the interaction of IL-4 and/or IL-13 with type 1 or type 2 receptors.

Non-limiting examples of classes of IL-4R antagonists include small molecule IL-4R inhibitors, anti-IL-4R aptamers, peptide-based IL-4R inhibitors (e.g., a "peptibody" molecule), a "receptor body" (e.g., an engineered molecule comprising a ligand binding domain of an IL-4R component), and antibodies or antigen-binding fragments of antibodies that specifically bind human IL-4Rα. As used herein, an IL-4R antagonist also includes an antigen binding protein that specifically binds IL-4 and/or IL-13.

Anti-IL-4 Rα antibodies and antigen binding fragments thereof

In certain exemplary embodiments of the present disclosure, the IL-4R antagonist is an anti-IL-4 Rα antibody or antigen-binding fragment thereof. As used herein, the term "antibody" includes immunoglobulin molecules comprising four polypeptide chains (two heavy (H) chains and two light (L) chains) connected to each other by disulfide bonds, and multimers thereof (e.g., igM). In a typical antibody, each heavy chain comprises a heavy chain variable region (abbreviated herein as HCVR or V _H) and a heavy chain constant region. The heavy chain constant region comprises three domains, C _H1、C_H and C _H. Each light chain comprises a light chain variable region (abbreviated herein as LCVR or V _L) and a light chain constant region. The light chain constant region comprises a domain (the C _L1).V_H region and the V _L region may be further subdivided into regions of hypervariability, known as Complementarity Determining Regions (CDRs), interspersed with regions that are more conserved, known as Framework Regions (FRs). Each of V _H and V _L consists of three CDRs and four FRs, from amino-terminus to carboxy-terminus, arranged in the order of FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. In some embodiments, the FRs of an anti-IL-4R antibody (or antigen binding portion thereof) are identical to human germline sequences.

As used herein, the term "antibody" also includes antigen binding fragments of whole antibody molecules. As used herein, the terms "antigen binding portion of an antibody," "antigen binding fragment of an antibody," and the like include any naturally occurring, enzymatically available, synthetic, or genetically engineered polypeptide or glycoprotein that specifically binds an antigen to form a complex. Antigen binding fragments of antibodies can be derived from whole antibody molecules, for example, using any suitable standard technique, such as proteolytic digestion or recombinant genetic engineering techniques involving manipulation and expression of DNA encoding antibody variable and optionally antibody constant domains. Such DNA is known and/or can be readily obtained from, for example, commercial sources, DNA libraries (including, for example, phage-antibody libraries), or can be synthesized. The DNA can be sequenced and manipulated chemically or by using molecular biological techniques, for example, to arrange one or more variable and/or constant domains into a suitable configuration, or to introduce codons, to create cysteine residues, to modify, add or delete amino acids, and the like.

Non-limiting examples of antigen binding fragments include (i) Fab fragments, (ii) F (ab') 2 fragments, (iii) Fd fragments, (iv) Fv fragments, (v) single chain Fv (scFv) molecules, (vi) dAb fragments, and (vii) minimal recognition units consisting of amino acid residues that mimic the hypervariable regions of an antibody (e.g., isolated Complementarity Determining Regions (CDRs) such as CDR3 peptides) or restricted FR3-CDR3-FR4 peptides. As used herein, other engineered molecules (such as domain-specific antibodies, single domain antibodies, domain deleted antibodies, chimeric antibodies, CDR-grafted antibodies, diabodies, trisomy, tetrasomy, microscler, nanobody (e.g., monovalent nanobody, bivalent nanobody, etc.), small Modular Immunopharmaceuticals (SMIP), and shark variable IgNAR domains) are also encompassed by the term "antigen-binding fragment".

The antigen binding fragment of an antibody typically comprises at least one variable domain. The variable domain may have any size or amino acid composition and will typically include at least one CDR contiguous to or in-frame with one or more framework sequences. In antigen binding fragments having a V _H domain associated with a V _L domain, the V _H and V _L domains can be positioned in any suitable arrangement relative to each other. For example, the variable region may be a dimer and contain V _H-V_H、V_H-V_L or V _L-V_L dimers. Alternatively, the antigen binding fragment of an antibody may contain the monomer V _H or V _L domain.

In certain embodiments, the antigen binding fragment of an antibody may contain at least one variable domain covalently linked to at least one constant domain. Non-limiting exemplary configurations of variable and constant domains that can be found within antigen binding fragments of antibodies of the present disclosure include ：(i)V_H-C_H1;(ii)V_H-C_H2;(iii)V_H-C_H3;(iv)V_H-C_H1-C_H2;(v)V_H-C_H1-C_H2-C_H3;(vi)V_H-C_H2-C_H3;(vii)V_H-C_L;(viii)V_L-C_H1;(ix)V_L-C_H2;(x)V_L-C_H3;(xi)V_L-C_H1-C_H2;(xii)V_L-C_H1-C_H2-C_H3;(xiii)V_L-C_H2-C_H3; and (xiv) V _L-C_L. In any configuration of variable and constant domains (including any of the exemplary configurations listed above), the variable and constant domains may be directly linked to each other or may be linked by full or partial hinge or linker regions. The hinge region may be comprised of at least 2 (e.g., 5, 10, 15, 20, 40, 60 or more) amino acids, which results in a flexible or semi-flexible linkage between adjacent variable and/or constant domains in a single polypeptide molecule. Furthermore, antigen binding fragments of antibodies of the present disclosure may comprise homodimers or heterodimers (or other multimers) having any of the variable domain and constant domain configurations listed above that are non-covalently associated with each other and/or with one or more monomer V _H or V _L domains (e.g., via disulfide bonds).

The constant region of an antibody is important for the ability of the antibody to fix complement and mediate cell-dependent cytotoxicity. Thus, in some embodiments, the isotype of the antibody may be selected based on whether antibody-mediated cytotoxicity requires the antibody.

As used herein, the term "antibody" also encompasses multispecific (e.g., bispecific) antibodies. The multispecific antibody or antigen-binding fragment of an antibody typically comprises at least two different variable domains, wherein each variable domain is capable of specifically binding to a separate antigen or a different epitope on the same antigen. Any multispecific antibody format may be suitable in the context of an antibody or antigen-binding fragment of an antibody of the present disclosure using conventional techniques available in the art. For example, in some embodiments, the methods of the present disclosure include the use of bispecific antibodies, wherein one arm of the immunoglobulin is specific for IL-4 ra or a fragment thereof and the other arm of the immunoglobulin is specific for a second therapeutic target or conjugated to a therapeutic moiety. Exemplary bispecific formats that may be used in the context of the present disclosure include, but are not limited to, scFv-based or diabody bispecific formats, igG-scFv fusions, double Variable Domain (DVD) -Ig, tetragenic hybridomas, knob-in holes, common light chains (e.g., common light chains with knob-in holes, etc.), crossMab, crossFab, (SEED) bodies, leucine zippers, duobodies, igG1/IgG2, dual-action Fab (DAF) -IgG, and Mab ² bispecific formats (for reviews of the foregoing formats see, e.g., klein et al 2012, mabs,4:6,1-11, and references cited therein). Bispecific antibodies can also be constructed using peptide/nucleic acid conjugation, for example, wherein unnatural amino acids with orthogonal chemical reactivity are used to generate site-specific antibody-oligonucleotide conjugates, which are then self-assembled into multimeric complexes of defined composition, valence, and geometry. (see, e.g., kazane et al, J.Am. Chem. Soc. [ electronic edition: 2012, 4 th month).

In some embodiments, the antibodies used in the methods of the present disclosure are human antibodies. As used herein, the term "human antibody" is intended to include antibodies having variable and constant regions derived from human germline immunoglobulin sequences. Nonetheless, the human antibodies of the present disclosure may comprise amino acid residues that are not encoded by human germline immunoglobulin sequences (e.g., mutations introduced by random or site-specific mutagenesis in vitro or by somatic mutation in vivo), for example in CDRs, and in particular in CDR 3. However, as used herein, the term "human antibody" is not intended to include antibodies in which CDR sequences derived from the germline of another mammalian species (such as a mouse) have been implanted onto a human framework sequence.

The antibodies used in the methods of the present disclosure may be recombinant human antibodies. As used herein, the term "recombinant human antibody" is intended to include all human antibodies prepared, expressed, established or isolated by recombinant means, such as antibodies expressed using recombinant expression vectors (described further below) transfected into host cells, antibodies isolated from recombinant combinatorial human antibody libraries (described further below), antibodies isolated from animals (e.g., mice) transgenic for human immunoglobulin genes (see, e.g., taylor et al, (1992) nucleic acids res., 20:6287-6295), or antibodies prepared, expressed, established or isolated by any other means that involves splicing human immunoglobulin gene sequences to other DNA sequences. Such recombinant human antibodies have variable and constant regions derived from human germline immunoglobulin sequences. However, in certain embodiments, such recombinant human antibodies undergo in vitro mutagenesis (or, when using animals that are transgenic for human Ig sequences, undergo in vivo somatic mutagenesis) and thus the amino acid sequences of the V _H and V _L regions of the recombinant antibodies are sequences that, although derived from and related to the human germline V _H and V _L sequences, may not naturally occur within the human antibody germline repertoire in vivo.

An "isolated antibody" refers to an antibody that has been identified and isolated and/or recovered from at least one component of a natural environment. For example, an antibody that has been isolated or removed from at least one component of an organism, or from a naturally occurring or naturally occurring tissue or cell of an antibody, is an "isolated antibody". Isolated antibodies also include in situ antibodies within recombinant cells. An isolated antibody is an antibody that has undergone at least one purification or isolation step. According to certain embodiments, the isolated antibody may be substantially free of other cellular material and/or chemicals.

According to certain embodiments, the antibodies used in the methods of the present disclosure specifically bind to IL-4 ra. As used herein, the term "specifically binds" as used herein means that an antibody or antigen-binding fragment thereof forms a complex with an antigen that is relatively stable under physiological conditions. Methods for determining whether an antibody specifically binds an antigen are well known in the art and include, for example, equilibrium dialysis, surface plasmon resonance, and the like. In some embodiments, an antibody that "specifically binds" IL-4 ra binds IL-4 ra with an equilibrium dissociation constant (K _D) of less than about 1000nM, less than about 500nM, less than about 300nM, less than about 200nM, less than about 100nM, less than about 90nM, less than about 80nM, less than about 70nM, less than about 60nM, less than about 50nM, less than about 40nM, less than about 30nM, less than about 20nM, less than about 10nM, less than about 5nM, less than about 1nM, less than about 0.5nM, less than about 0.25nM, less than about 0.1nM, or less than about 0.05nM to IL-4 ra or a portion thereof, as measured in a surface plasmon resonance assay (e.g., BIAcore LIFE SCIENCES division of BIAcore ^TM, GE HEALTHCARE, piscataway, NJ). In some embodiments, antibodies that specifically bind to a target antigen (e.g., IL-4Rα) may also specifically bind to another antigen, such as an ortholog of the target antigen. For example, in some embodiments, an isolated antibody that specifically binds human IL-4rα exhibits cross-reactivity to other antigens, such as IL-4rα molecules from other (non-human) species.

In some embodiments, the IL-4R antagonist is an anti-IL-4R alpha antibody or antigen-binding fragment thereof comprising a Heavy Chain Variable Region (HCVR), a Light Chain Variable Region (LCVR), and/or a Complementarity Determining Region (CDR) comprising any of the amino acid sequences of an anti-IL-4R antibody described in U.S. patent No. 7,608,693, incorporated herein by reference. In some embodiments, the IL-4R antagonist is an anti-IL-4R alpha antibody or antigen-binding fragment thereof comprising a heavy chain complementarity determining region (HCDR) comprising a Heavy Chain Variable Region (HCVR) of the amino acid sequence of SEQ ID NO. 1 and a light chain complementarity determining region (LCDR) comprising a Light Chain Variable Region (LCVR) of the amino acid sequence of SEQ ID NO. 2. In some embodiments, the IL-4R antagonist is an anti-IL-4R alpha antibody or antigen binding fragment thereof comprising three HCDRs (HCDR 1, HCDR2, and HCDR 3) and three LCDRs (LCDR 1, LCDR2, and LCDR 3), wherein HCDR1 comprises amino acid sequence GFTFRDYA (SEQ ID NO: 3), HCDR2 comprises amino acid sequence ISGSGGNT (SEQ ID NO: 4), HCDR3 comprises amino acid sequence AKDRLSITIRPRYYGLDV (SEQ ID NO: 5), LCDR1 comprises amino acid sequence QSLLYSIGYNY (SEQ ID NO: 6), LCDR2 comprises amino acid sequence LGS, and LCDR3 comprises amino acid sequence MQALQTPYT (SEQ ID NO: 8).

In some embodiments, an anti-IL-4R antibody or antigen-binding fragment thereof comprises HCDR1 comprising amino acid sequence GFTFRDYA (SEQ ID NO: 3), HCDR2 comprising amino acid sequence ISGSGGNT (SEQ ID NO: 4), HCDR3 comprising amino acid sequence AKDRLSITIRPRYYGLDV (SEQ ID NO: 5), LCDR1 comprising amino acid sequence QSLLYSIGYNY (SEQ ID NO: 6), LCDR2 comprising amino acid sequence LGS, and LCDR3 comprising amino acid sequence MQALQTPYT (SEQ ID NO: 8), and further comprises a HCVR having at least 85% sequence identity (e.g., at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity) to the amino acid sequence of SEQ ID NO:1, and a LCVR having at least 85% sequence identity (e.g., at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity) to the amino acid sequence of SEQ ID NO: 2. In some embodiments, the anti-IL-4R antibody or antigen-binding fragment thereof comprises a HCVR comprising SEQ ID NO. 1 and a LCVR comprising SEQ ID NO. 2.

In some embodiments, the anti-IL-4R antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO. 9. In some embodiments, the anti-IL-4R antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO. 10.

An exemplary antibody comprising a heavy chain comprising the amino acid sequence of SEQ ID NO. 9 and a light chain comprising the amino acid sequence of SEQ ID NO. 10 is a fully human anti-IL-4R antibody known as Dupu Li Youshan antibody. According to certain exemplary embodiments, the methods of the present disclosure include the use of a duplicon Li Youshan antibody. As used herein, "dupu Li Youshan antibody" also includes bioequivalence of dupup Li Youshan antibody. The term "bioequivalence" as used herein with respect to the duplicon Li Youshan antibody refers to an anti-IL-4R antibody or IL-4R binding protein or fragment thereof as a pharmaceutical equivalent or pharmaceutical substitute that does not exhibit a significant difference in rate and/or extent of absorption from duplicon Li Youshan antibody when administered in the same molar dose (whether single or multiple doses) under similar experimental conditions. In some embodiments, the term refers to an antigen binding protein that binds to IL-4R that does not have clinically significant differences in its safety, purity, and/or potency from the dopen Li Youshan antibody.

Other anti-IL-4 Rα antibodies that may be used in the context of the methods of the present disclosure include, for example, antibodies known in the art as AMG317 (Corren et al, 2010,Am J Respir Crit Care Med, 181 (8): 788-796), or MEDI 9314, or any of the anti-IL-4 Rα antibodies described in U.S. Pat. No. 7,186,809, U.S. Pat. No. 7,605,237, U.S. Pat. No. 7,638,606, U.S. Pat. No. 8,092,804, U.S. Pat. No. 8,679,487, U.S. Pat. No. 8,877,189, U.S. Pat. No. 10,774,141, or International patent publication No. WO2020/096381, WO 2020/182197, WO2020/239134, WO 2021/213329, WO2022/052974, WO2022/136669, or WO2022/136675, the contents of each of which are incorporated herein by reference.

In some embodiments, an anti-IL-4 ra antibody or antigen-binding fragment thereof for use in the methods of the present disclosure comprises one or more of the CDR, HCVR, and/or LCVR sequences listed in table 1 below.

In some embodiments, the anti-IL-4 Rα antibody comprises (i) a HCVR comprising the amino acid sequence SEQ ID NO:32(SCB-VH-59)、SEQ ID NO:33(SCB-VH-60)、SEQ ID NO:34(SCB-VH-61)、SEQ ID NO:35(SCB-VH-62)、SEQ ID NO:36(SCB-VH-63)、SEQ ID NO:37(SCB-VH-64)、SEQ ID NO:38(SCB-VH-65)、SEQ ID NO:39(SCB-VH-66)、SEQ ID NO:40(SCB-VH-67)、SEQ ID NO:41(SCB-VH-68)、SEQ ID NO:42(SCB-VH-69)、SEQ ID NO:43(SCB-VH-70)、SEQ ID NO:44(SCB-VH-71)、SEQ ID NO:45(SCB-VH-72)、SEQ ID NO:46(SCB-VH-73)、SEQ ID NO:47(SCB-VH-74)、SEQ ID NO:48(SCB-VH-75)、SEQ ID NO:49(SCB-VH-76)、SEQ ID NO:50(SCB-VH-77)、SEQ ID NO:51(SCB-VH-78)、SEQ ID NO:52(SCB-VH-79)、SEQ ID NO:53(SCB-VH-80)、SEQ ID NO:54(SCB-VH-81)、SEQ ID NO:55(SCB-VH-82)、SEQ ID NO:56(SCB-VH-83)、SEQ ID NO:57(SCB-VH-84)、SEQ ID NO:58(SCB-VH-85)、SEQ ID NO:59(SCB-VH-86)、SEQ ID NO:60(SCB-VH-87)、SEQ ID NO:61(SCB-VH-88)、SEQ ID NO:62(SCB-VH-89)、SEQ ID NO:63(SCB-VH-90)、SEQ ID NO:64(SCB-VH-91)、SEQ ID NO:65(SCB-VH-92) or SEQ ID NO:66 (SCB-VH-93), and (ii) a LCVR comprising the amino acid sequence SEQ ID NO:12(SCB-VL-39)、SEQ ID NO:13(SCB-VL-40)、SEQ ID NO:14(SCB-VL-41)、SEQ ID NO:15(SCB-VL-42)、SEQ ID NO:16(SCB-VL-43)、SEQ ID NO:17(SCB-VL-44)、SEQ ID NO:18(SCB-VL-45)、SEQ ID NO:19(SCB-VL-46)、SEQ ID NO:20(SCB-VL-47)、SEQ ID NO:21(SCB-VL-48)、SEQ ID NO:22(SCB-VL-49)、SEQ ID NO:23(SCB-VL-50)、SEQ ID NO:24(SCB-VL-51)、SEQ ID NO:25(SCB-VL-52)、SEQ ID NO:26(SCB-VL-53)、SEQ ID NO:27(SCB-VL-54)、SEQ ID NO:28(SCB-VL-55)、SEQ ID NO:29(SCB-VL-56)、SEQ ID NO:30(SCB-VL-57) or SEQ ID NO:31 (SCB-VL-58). In some embodiments, the anti-IL-4 Rα antibody comprises a HCVR comprising the amino acid sequence of SEQ ID NO:64 (SCB-VH-91) and a LCVR comprising the amino acid sequence of SEQ ID NO:17 (SCB-VL-44), SEQ ID NO:27 (SCB-VL-54) or SEQ ID NO:28 (SCB-VL-55).

In some embodiments, the anti-IL-4 Rα antibody comprises an amino acid sequence pair ：SEQ ID NO:67/68(MEDI-1-VH/MEDI-1-VL);SEQ ID NO:69/70(MEDI-2-VH/MEDI-2-VL);SEQ ID NO:71/72(MEDI-3-VH/MEDI-3-VL);SEQ ID NO:73/74(MEDI-4-VH/MEDI-4-VL);SEQ ID NO:75/76(MEDI-5-VH/MEDI-5-VL);SEQ ID NO:77/78(MEDI-6-VH/MEDI-6/VL);SEQ ID NO:79/80(MEDI-7-VH/MEDI-7-VL);SEQ ID NO:81/82(MEDI-8-VH/MEDI-8-VL);SEQ ID NO:83/84(MEDI-9-VH/MEDI-9-VL);SEQ ID NO:85/86(MEDI-10-VH/MEDI-10-VL);SEQ ID NO:87/88(MEDI-11-VH/MEDI-11/VL);SEQ ID NO:89/90(MEDI-12-VH/MEDI-12-VL);SEQ ID NO:91/92(MEDI-13-VH/MEDI-13-VL);SEQ ID NO:93/94(MEDI-14-VH/MEDI-14-VL);SEQ ID NO:95/96(MEDI-15-VH/MEDI-15-VL);SEQ ID NO:97/98(MEDI-16-VH/MEDI-16/VL);SEQ ID NO:99/100(MEDI-17-VH/MEDI-17-VL);SEQ ID NO:101/102(MEDI-18-VH/MEDI-18-VL);SEQ ID NO:103/104(MEDI-19-VH/MEDI-19-VL);SEQ ID NO:105/106(MEDI-20-VH/MEDI-20-VL);SEQ ID NO:107/108(MEDI-21-VH/MEDI-21-VL);SEQ ID NO:109/110(MEDI-22-VH/MEDI-22-VL);SEQ ID NO:111/112(MEDI-23-VH/MEDI-23-VL);SEQ ID NO:113/114(MEDI-24-VH/MEDI-24-VL);SEQ ID NO:115/116(MEDI-25-VH/MEDI-25-VL);SEQ ID NO:117/118(MEDI-26-VH/MEDI-26-VL);SEQ ID NO:119/120(MEDI-27-VH/MEDI-27-VL);SEQ ID NO:121/122(MEDI-28-VH/MEDI-28-VL);SEQ ID NO:123/124(MEDI-29-VH/MEDI-29-VL);SEQ ID NO:125/126(MEDI-30-VH/MEDI-30-VL);SEQ ID NO:127/128(MEDI-31-VH/MEDI-31-VL);SEQ ID NO:129/130(MEDI-32-VH/MEDI-32-VL);SEQ ID NO:131/132(MEDI-33-VH/MEDI-33-VL);SEQ ID NO:133/134(MEDI-34-VH/MEDI-34-VL);SEQ ID NO:135/136(MEDI-35-VH/MEDI-35-VL);SEQ ID NO:137/138(MEDI-36-VH/MEDI-36-VL);SEQ ID NO:139/140(MEDI-37-VH/MEDI-37-VL);SEQ ID NO:141/142(MEDI-38-VH/MEDI-38-VL);SEQ ID NO:143/144(MEDI-39-VH/MEDI-39-VL);SEQ ID NO:145/146(MEDI-40-VH/MEDI-40-VL);SEQ ID NO:147/148(MEDI-41-VH/MEDI-41-VL);SEQ ID NO:149/150(MEDI-42-VH/MEDI-42-VL); selected from the group consisting of and SEQ ID NO 151/152 (MEDI-37 GL-VH/MEDI-37 GL-VL).

In some embodiments, the anti-IL-4 Rα antibody comprises (i) a HCVR comprising the amino acid sequence SEQ ID NO:153(AJOU-1-VH)、SEQ ID NO:154(AJOU-2-VH)、SEQ ID NO:155(AJOU-3-VH)、SEQ ID NO:156(AJOU-4-VH)、SEQ ID NO:157(AJOU-5-VH)、SEQ ID NO:158(AJOU-6-VH)、SEQ ID NO:159(AJOU-7-VH)、SEQ ID NO:160(AJOU-8-VH)、SEQ ID NO:161(AJOU-9-VH)、SEQ ID NO:162(AJOU-10-VH)、SEQ ID NO:163(AJOU-69-VH)、SEQ ID NO:164(AJOU-70-VH)、SEQ ID NO:165(AJOU-71-VH)、SEQ ID NO:166(AJOU-72-VH) or SEQ ID NO:167 (AJOU-83-VH), and (ii) a LCVR comprising the amino acid sequence SEQ ID NO:168(AJOU-33-VL)、SEQ ID NO:169(AJOU-34-VL)、SEQ ID NO:170(AJOU-35-VL)、SEQ ID NO:171(AJOU-36-VL)、SEQ ID NO:172(AJOU-37-VL)、SEQ ID NO:173(AJOU-38-VL)、SEQ ID NO:174(AJOU-39-VL)、SEQ ID NO:175(AJOU-40-VL)、SEQ ID NO:176(AJOU-41-VL)、SEQ ID NO:177(AJOU-42-VL)、SEQ ID NO:178(AJOU-77-VL)、SEQ ID NO:179(AJOU-78-VL)、SEQ ID NO:180(AJOU-79-VL)、SEQ ID NO:181(AJOU-80-VL)、SEQ ID NO:182(AJOU-86-VL)、SEQ ID NO:183(AJOU-87-VL)、SEQ ID NO:184(AJOU-88-VL)、SEQ ID NO:185(AJOU-89-VL)、SEQ ID NO:186(AJOU-90-VL) or SEQ ID NO:187 (AJOU-91-VL).

In some embodiments, the anti-IL-4Rα antibody comprises (i) a HCVR comprising the amino acid sequence SEQ ID NO:188(REGN-VH-3)、SEQ ID NO:189(REGN-VH-19)、SEQ ID NO:190(REGN-VH-35)、SEQ ID NO:191(REGN-VH-51)、SEQ ID NO:192(REGN-VH-67)、SEQ ID NO:193(REGN-VH-83)、SEQ ID NO:194(REGN-VH-99)、SEQ ID NO:195(REGN-VH-115)、SEQ ID NO:196(REGN-VH-147) or SEQ ID NO 197 (REGN-VH-163), and (ii) a LCVR comprising the amino acid sequence SEQ ID NO:198(REGN-VL-11)、SEQ ID NO:199(REGN-VL-27)、SEQ ID NO:200(REGN-VL-43)、SEQ ID NO:201(REGN-VL-59)、SEQ ID NO:202(REGN-VL-75)、SEQ ID NO:203(REGN-VL-91)、SEQ ID NO:204(REGN-VL-107)、SEQ ID NO:205(REGN-VL-123)、SEQ ID NO:206(REGN-VL-155) or SEQ ID NO 207 (REGN-VL-171).

In some embodiments, the anti-IL-4Rα antibody comprises (i) a HCVR comprising the amino acid sequence SEQ ID NO:208(STSA-C27-VH)、SEQ ID NO:209(STSA-C27-6-33-VH)、SEQ ID NO:210(STSA-C27-7-33-VH)、SEQ ID NO:211(STSA-C27-24-56-VH)、SEQ ID NO:212(STSA-C27-47-56-VH)、SEQ ID NO:213(STSA-C27-33-33-VH)、SEQ ID NO:214(STSA-C27-56-56-VH)、SEQ ID NO:215(STSA-C27-78-78-VH)、SEQ ID NO:216(STSA-C27-82-58-VH)、SEQ ID NO:217(STSA-C27-54-54-VH)、SEQ ID NO:218(STSA-C27-36-36-VH)、SEQ ID NO:219(STSA-C27-53-53-VH)、SEQ ID NO:220(STSA-C27-67-67-VH)、SEQ ID NO:221(STSA-C27-55-55-VH)、SEQ ID NO:222(STSA-C27-59-59-VH)、SEQ ID NO:223(STSA-C27-58-58-VH)、SEQ ID NO:224(STSA-C27-52-52-VH) or SEQ ID NO:225 (STSA-C27-Y2-Y2-VH), and (ii) a LCVR comprising the amino acid sequence SEQ ID NO:226(STSA-C27-VL)、SEQ ID NO:227(STSA-C27-6-33-VL)、SEQ ID NO:228(STSA-C27-7-33-VL)、SEQ ID NO:229(STSA-C27-24-56-VL)、SEQ ID NO:230(STSA-C27-47-56-VL)、SEQ ID NO:231(STSA-C27-33-33-VL)、SEQ ID NO:232(STSA-C27-56-56-VL)、SEQ ID NO:233(STSA-C27-78-78-VL)、SEQ ID NO:234(STSA-C27-82-58-VL)、SEQ ID NO:235(STSA-C27-54-54-VL)、SEQ ID NO:236(STSA-C27-36-36-VL)、SEQ ID NO:237(STSA-C27-53-53-VL)、SEQ ID NO:238(STSA-C27-67-67-VL)、SEQ ID NO:239(STSA-C27-55-55-VL)、SEQ ID NO:240(STSA-C27-59-59-VL)、SEQ ID NO:241(STSA-C27-58-58-VL)、SEQ ID NO:242(STSA-C27-52-52-VL) or SEQ ID NO:243 (STSA-C27-Y2-Y2-VL).

In some embodiments, the anti-IL-4 Rα antibody comprises (i) a HCVR comprising the amino acid sequence SEQ ID NO:244(Y0188-1 VH)、SEQ ID NO:245(Y0188-2 VH)、SEQ ID NO:246(Y0188-3 VH)、SEQ ID NO:247(Y0188-4 VH)、SEQ ID NO:248(Y0188-6VH)、SEQ ID NO:249(Y0188-8 VH)、SEQ ID NO:250(Y0188-9 VH)、SEQ ID NO:251(Y0188-10 VH)、SEQ ID NO:252(Y0188-14 VH)、SEQ ID NO:253(HV3-15-14VH)、SEQ ID NO:254(HV3-48-14VH)、SEQ ID NO:255(HV3-73*2-14VH)、SEQ ID NO:256(HV3-72-14VH)、SEQ ID NO:257(Y01-14 VH)、SEQ ID NO:258(162-14VH) or SEQ ID NO 259 (VH 73-14 Vh) and (ii) a LCVR comprising the amino acid sequence SEQ ID NO:260(Y0188-1 VL)、SEQ ID NO:261(Y0188-2 VL)、SEQ ID NO:262(Y0188-3 VL)、SEQ ID NO:263(Y0188-4 VL)、SEQ ID NO:264(Y0188-6 VL)、SEQ ID NO:265(Y0188-8VL)、SEQ ID NO:266(Y0188-9 VL)、SEQ ID NO:267(Y0188-10 VL)、SEQ ID NO:268(Y0188-14 VL)、SEQ ID NO:269(Y01-14 VL)、SEQ ID NO:270(164-14VL)、SEQ ID NO:271(KV4-14 VL)、SEQ ID NO:272(KV1-27-14VL)、SEQ ID NO:273(KV1-9-14VL)、SEQ ID NO:274(KV1-NL1-14 VL) or SEQ ID NO 275 (KV 1D-43-14 VL).

In some embodiments, the anti-IL-4 ra antibodies used in the methods of the present disclosure may have pH-dependent binding characteristics. For example, an anti-IL-4Rα antibody as used herein disclosed may exhibit reduced binding to IL-4Rα at acidic pH as compared to neutral pH. Alternatively, an anti-IL-4 ra antibody as used herein disclosed may exhibit enhanced binding to its antigen at acidic pH compared to neutral pH. The expression "acidic pH" includes pH values of less than about 6.2, such as about 6.0, 5.95, 5.9, 5.85, 5.8, 5.75, 5.7, 5.65, 5.6, 5.55, 5.5, 5.45, 5.4, 5.35, 5.3, 5.25, 5.2, 5.15, 5.1, 5.05, 5.0 or less. As used herein, the expression "neutral pH" refers to a pH of about 7.0 to about 7.4. The expression "neutral pH" includes pH values of about 7.0, 7.05, 7.1, 7.15, 7.2, 7.25, 7.3, 7.35 and 7.4.

In some cases, "reduced binding to IL-4 ra at acidic pH compared to neutral pH" is expressed as the ratio of the K _D value of binding of the antibody to IL-4 ra at acidic pH to the K _D value of binding of the antibody to IL-4 ra at neutral pH (or vice versa). For example, for the purposes of this disclosure, an antibody or antigen-binding fragment thereof may be considered to exhibit "reduced binding to IL-4 ra at acidic pH compared to neutral pH" if the antibody or antigen-binding fragment thereof exhibits an acidic/neutral K _D ratio of about 3.0 or greater. In certain exemplary embodiments, the acid/neutral K _D ratio of an antibody or antigen-binding fragment of the present disclosure may be about 3.0、3.5、4.0、4.5、5.0、5.5、6.0、6.5、7.0、7.5、8.0、8.5、9.0、9.5、10.0、10.5、11.0、11.5、12.0、12.5、13.0、13.5、14.0、14.5、15.0、20.0、25.0、30.0、40.0、50.0、60.0、70.0、100.0 or greater.

Antibodies with pH-dependent binding characteristics can be obtained, for example, by screening a population of antibodies for reduced (or enhanced) binding to a particular antigen at an acidic pH as compared to a neutral pH. In addition, modification of the antigen binding domain at the amino acid level may result in antibodies with pH-dependent characteristics. For example, by replacing one or more amino acids of the antigen binding domain (e.g., within a CDR) with histidine residues, antibodies can be obtained that have reduced antigen binding at acidic pH relative to neutral pH.

Preparation of human antibodies

Methods for producing human antibodies in transgenic mice are known in the art. Any such known methods may be used in the context of the present disclosure to prepare human antibodies that specifically bind to human IL-4R.

The high affinity chimeric antibody against IL-4R with human variable and mouse constant regions is initially isolated using VELOCIMMUNE ^TM techniques (see, e.g., US 6,596,541, regenerator pharmaceutical company (Regeneron Pharmaceuticals)) or any other known method for producing monoclonal antibodies.The technology relates to generating transgenic mice having genomes comprising human heavy and light chain variable regions operably linked to endogenous mouse constant region loci such that the mice produce antibodies comprising human variable regions and mouse constant regions in response to antigen stimulation. DNA encoding the heavy and light chain variable regions of the antibody is isolated and operably linked to DNA encoding the human heavy and light chain constant regions. Subsequently, the DNA is expressed in cells capable of expressing fully human antibodies.

Typically, challenge with antigen of interestMice, and lymphocytes (such as B cells) are recovered from the mice expressing the antibodies. Lymphocytes can be fused with myeloma cell lines to produce immortalized hybridoma cell lines, and such hybridoma cell lines are screened and selected to identify hybridoma cell lines that produce antibodies specific for the antigen of interest. DNA encoding the heavy and light chain variable regions can be isolated and linked to the desired heavy and light chain isotype constant regions. Such antibody proteins may be produced in cells such as CHO cells. Alternatively, DNA encoding the antigen-specific chimeric antibodies or the light and heavy chain variable domains may be isolated directly from antigen-specific lymphocytes.

First, a high affinity chimeric antibody having a human variable region and a mouse constant region was isolated. Antibodies are characterized and selected to obtain desired characteristics, including affinity, selectivity, epitope, etc., using standard procedures known to those skilled in the art. The mouse constant region is replaced with the desired human constant region to produce a fully human antibody of the present disclosure, e.g., wild-type or modified IgG1 or IgG4. While the constant region selected may vary depending on the particular application, high affinity antigen binding and target-specific features are present in the variable region.

In general, antibodies useful in the methods of the present disclosure have high affinity as described above when measured by binding to an antigen immobilized on a solid phase or in a solution phase. The mouse constant region is replaced with the desired human constant region to produce the fully human antibodies of the present disclosure. While the constant region selected may vary depending on the particular application, high affinity antigen binding and target-specific features are present in the variable region.

In one embodiment, a human antibody or antigen-binding fragment thereof that specifically binds IL-4R and is useful in the methods disclosed herein comprises three heavy chain CDRs (HCDR 1, HCDR2, and HCDR 3) contained within a Heavy Chain Variable Region (HCVR) having the amino acid sequence of SEQ ID NO. 1 and three light chain CDRs (LCVR 1, LCVR2, and LCVR 3) contained within a Light Chain Variable Region (LCVR) having the amino acid sequence of SEQ ID NO. 2. Methods and techniques for identifying CDRs within HCVR and LCVR amino acid sequences are well known in the art and can be used to identify CDRs within a given HCVR and/or LCVR amino acid sequence disclosed herein. Exemplary conventions that may be used to identify boundaries of CDRs include, for example, kabat definitions, chothia definitions, and AbM definitions. In general, kabat definition is based on sequence differences, chothia definition is based on the position of structural loop regions, and AbM definition is a compromise between Kabat and Chothia methods. See, e.g., ,Kabat,"Sequences of Proteins of Immunological Interest,"National Institutes of Health,Bethesda,Md.(1991);Al-Lazikani et al, J.mol.biol.,273:927-948 (1997), and Martin et al, proc.Natl.Acad.Sci.USA,86:9268-9272 (1989). Public databases can also be used to identify CDR sequences within antibodies.

Pharmaceutical composition

In one aspect, the present disclosure provides methods comprising administering an IL-4R antagonist to a subject, wherein the IL-4R antagonist (e.g., an anti-IL-4R antibody) is contained in a pharmaceutical composition comprising one or more pharmaceutically acceptable vehicles, carriers, and/or excipients. Various pharmaceutically acceptable carriers and excipients are well known in the art. See, e.g., remington's Pharmaceutical Sciences, mack Publishing Company, easton, PA. In some embodiments, the carrier is suitable for intravenous, intramuscular, oral, intraperitoneal, intrathecal, transdermal, topical, or subcutaneous administration.

Methods of administration include, but are not limited to, intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, epidural, and oral routes. The compositions may be administered by any convenient route, for example by infusion or bolus injection, by absorption through the epithelial or mucocutaneous linings (e.g., oral mucosa, rectal and intestinal mucosa, etc.), and may be administered with other bioactive agents. In some embodiments, the pharmaceutical compositions as disclosed herein are administered intravenously. In some embodiments, the pharmaceutical compositions as disclosed herein are administered subcutaneously.

In some embodiments, the pharmaceutical composition comprises an injectable formulation, such as a dosage form for intravenous, subcutaneous, intradermal, and intramuscular injection, instillation, and the like. These injectable formulations can be prepared by known methods. For example, injectable formulations can be prepared, for example, by dissolving, suspending or emulsifying the antibodies or salts thereof described above in sterile aqueous or oily media conventionally used for injection. As the aqueous medium for injection, there are, for example, physiological saline, isotonic solution containing glucose and other auxiliary agents, etc., which can be used in combination with an appropriate solubilizing agent such as alcohol (e.g., ethanol), polyol (e.g., propylene glycol, polyethylene glycol), nonionic surfactant [ e.g., polysorbate 80, HCO-50 (polyoxyethylene (50 mol) adduct of hydrogenated castor oil) ] and the like. As the oily medium, for example, sesame oil, soybean oil, etc., are used, and they may be used in combination with a solubilizing agent such as benzyl benzoate, benzyl alcohol, etc. The injection thus prepared may be filled into a suitable ampoule.

The dosage of antibody administered to a subject according to the methods of the present disclosure may vary depending on the age and size of the subject, symptoms, conditions, route of administration, and the like. The dose is typically calculated from body weight or body surface area. Depending on the severity of the condition, the frequency and duration of treatment may be adjusted. The effective dosage and schedule for administration of a pharmaceutical composition comprising an anti-IL-4R antibody may be determined empirically, for example, the progress of the patient may be monitored by periodic assessment and the dosage adjusted accordingly. In addition, dose inter-species scaling may be performed using methods well known in the art (e.g., mordenti et al, 1991, pharmaceutiut. Res.,8: 1351). Specific exemplary dosages of anti-IL 4R antibodies useful in the context of the present disclosure and administration regimens involving such dosages are disclosed elsewhere herein.

In some embodiments, the IL-4R antagonists or pharmaceutical compositions of the disclosure are contained in a container. Thus, in another aspect, there is provided a container comprising an IL-4R antagonist or pharmaceutical composition as disclosed herein. For example, in some embodiments, the pharmaceutical composition is contained within a container selected from the group consisting of a glass vial, a syringe, a pen delivery device, and an auto-injector.

In some embodiments, the pharmaceutical compositions of the present disclosure are delivered using standard needles and syringes, e.g., subcutaneously or intravenously. In some embodiments, the syringe is a pre-filled syringe. In some embodiments, a pen delivery device or an auto-injector is used to deliver the pharmaceutical compositions of the present disclosure (e.g., for subcutaneous delivery). The pen delivery device may be reusable or disposable. Typically, reusable pen delivery devices utilize replaceable cartridges containing the pharmaceutical composition. Once the pharmaceutical composition within the cartridge has been administered and the cartridge is empty, the empty cartridge can be easily discarded and replaced with a new cartridge containing the pharmaceutical composition. The pen delivery device may then be reused. In disposable pen delivery devices, there is no replaceable cartridge. In practice, disposable pen delivery devices are prefilled with a pharmaceutical composition in a reservoir within the device. Once the reservoir is emptied of the pharmaceutical composition, the entire device is discarded.

Examples of suitable pen and auto-injector delivery devices include, but are not limited to AUTOPEN ^TM(Owen Mumford,Inc.,Woodstock,UK)、DISETRONIC^TM pens (Disetronic MEDICAL SYSTEMS, bergdorf, switzerland), HUMALOG MIX 75/25 ^TM pens, HUMALOG ^TM pens, HUMALIN 70/30 ^TM pen (ELI LILLY AND Co., indianapolis, ind.), NOVOPEN ^TM I, II and III(Novo Nordisk,Copenhagen,Denmark)、NOVOPEN JUNIOR^TM(Novo Nordisk,Copenhagen,Denmark)、BD^TM pen (Becton Dickinson, FRANKLIN LAKES, NJ), OPTIPEN ^TM、OPTIPEN PRO^TM、OPTIPEN STARLET^TM and OPTICLIK ^TM (sanofi-aventis, frankfurt, germany). Examples of disposable pen delivery devices useful for subcutaneous delivery of the pharmaceutical compositions of the present disclosure include, but are not limited to, SOLOSTAR ^TM pens (sanofi-aventis), FLEXPEN ^TM (Novo Nordisk) and KWIKPEN ^TM(Eli Lilly)、SURECLICK^TM auto-injectors (Amgen, thround Oaks, calif.), a pen, PENLET ^TM (HASELMEIER, stuttgart, germany), EPIPEN (Dey, L.P.), and HUMIRA ^TM pens (Abbott Labs, abbott Park IL).

In some embodiments, the pharmaceutical composition is delivered using a controlled release system. In one embodiment, a pump (see Langer, supra; sefton,1987,CRC Crit.Ref.Biomed.Eng.14:201) may be used. In another embodiment, polymeric materials may be used, see Medical Applications of Controlled Release, langer and Wise (ed.), 1974, CRC Pres., boca Raton, florida. In yet another embodiment, the controlled release system may be placed in proximity to the composition target, thus requiring only a small portion of the systemic dose (see, e.g., goodson,1984, supra, medical Applications of Controlled Release, volume 2, pages 115-138). Other controlled release systems are discussed in the reviews by Langer,1990, science, 249:1527-1533. Other delivery systems are known and may be used to administer pharmaceutical compositions, for example encapsulated in liposomes, microparticles, microcapsules, recombinant cells capable of expressing mutant viruses, receptor-mediated endocytosis (see e.g. Wu et al, 1987, j. Biol. Chem., 262:4429-4432).

In some embodiments, the pharmaceutical composition comprising an anti-IL-4R antibody is administered using a drug delivery device that is a needle-based injection system, as described in table 1 of section 5.2 of ISO 11608-1:2014 (E). Needle-based injection systems can be broadly divided into multi-dose container systems and single-dose (partially or fully evacuated) container systems, as described in ISO 11608-1:2014 (E). The container may be a replaceable container or an integral non-replaceable container.

As further described in ISO 11608-1:2014 (E), a multi-dose container system may involve a needle-based injection device with a replaceable container. In such a system, each container contains multiple doses, which may be of fixed or variable size (preset by the user). Another multi-dose container system may involve a needle-based injection device with an integrated non-replaceable container. In such a system, each container contains multiple doses, which may be of fixed or variable size (preset by the user).

As further described in ISO 11608-1:2014 (E), single dose container systems may involve needle-based injection devices with replaceable containers. In one example of such a system, each container contains a single dose, expelling the entire deliverable volume (fully evacuated). In another example, each container contains a single dose, expelling a portion of the deliverable volume (partial evacuation). Further as described in ISO 11608-1:2014 (E), single dose container systems may involve needle-based injection devices with integrated non-exchangeable containers. In one example of such a system, each container contains a single dose, expelling the entire deliverable volume (fully evacuated). In another example, each container contains a single dose, expelling a portion of the deliverable volume (partial evacuation).

An exemplary sleeve-triggered auto-injector with a manual needle insert is described in International publication WO 2015/004052. Exemplary audible end-of-dose feedback mechanisms are described in International publications WO2016/193346 and WO 2016/193348. An exemplary needle safety mechanism after use of an automatic injector is described in International publication WO 2016/193352. An exemplary needle sheath remover mechanism for an automatic injector for a syringe is described in International publication WO 2016/193353. An exemplary support mechanism for supporting the axial position of the syringe is described in International publication WO 2016/193355.

In some embodiments, the pharmaceutical compositions for use as described herein are prepared in dosage forms in unit doses appropriate for the dosage of the active ingredient. Such dosage forms in unit dosage form include, for example, tablets, pills, capsules, injectable solutions (ampoules), suppositories and the like.

Exemplary pharmaceutical compositions comprising anti-IL-4R antibodies that can be used in the context of the present disclosure are disclosed, for example, in U.S. patent 8,945,559.

Dosage and administration

In some embodiments, a therapeutically effective amount of an IL-4R antagonist (e.g., an anti-IL-4R antibody) is administered to a subject (e.g., a subject with a defect in bone growth) according to the methods of the present disclosure. As used herein with respect to an IL-4R antagonist, the phrase "therapeutically effective amount" refers to an amount of an IL-4R antagonist that results in one or more of (a) an improvement in bone formation, (b) an improvement in bone mineralization and/or bone mineral density, (c) a reduction in bone loss, (d) an improvement or normalization of one or more biomarkers of bone formation or bone turnover (such as, but not limited to, bone-specific alkaline phosphatase, carboxy-terminal cross-terminal peptide of type I collagen, N-terminal pro-peptide of type I procollagen, insulin-like growth factor 1, or osteocalcin) (e.g., relative to a healthy control value), and/or (e) a reduction in bone mass, osteoporosis, or reduced incidence of bone fracture (e.g., relative to a healthy control value).

In the case of anti-IL-4R antibodies, a therapeutically effective amount may be from about 0.05mg to about 600mg, e.g., about 0.05mg, about 0.1mg, about 1.0mg, about 1.5mg, about 2.0mg, about 10mg, about 20mg, about 30mg, about 40mg, about 50mg, about 60mg, about 70mg, about 80mg, about 90mg, about 100mg, about 110mg, about 120mg, about 130mg, about 140mg, about 150mg, about 160mg, about 170mg, about 180mg, about 190mg, about 200mg, about 210mg, about 220mg, about 230mg, about 240mg, about 250mg, about 260mg, about 270mg, about 280mg, about 290mg about 300mg, about 310mg, about 320mg, about 330mg, about 340mg, about 350mg, about 360mg, about 370mg, about 380mg, about 390mg, about 400mg, about 410mg, about 420mg, about 430mg, about 440mg, about 450mg, about 460mg, about 470mg, about 480mg, about 490mg, about 500mg, about 510mg, about 520mg, about 530mg, about 540mg, about 550mg, about 560mg, about 570mg, about 580mg, about 590mg, or about 600mg of the anti-IL-4R antibody. In some embodiments, the therapeutically effective amount is from about 50mg to about 600mg, or from about 100mg to about 600mg, or from about 200mg to about 600mg. In certain embodiments, 50mg, 75mg, 100mg, 125mg, 150mg, 200mg, 250mg, 300mg, 350mg, 400mg, 450mg, 500mg, 550mg, or 600mg of an anti-IL-4R antibody is administered to a subject.

The amount of IL-4R antagonist (e.g., anti-IL-4R antibody) contained in an individual dose can be expressed in milligrams of antibody per kilogram of subject body weight (i.e., mg/kg). For example, the IL-4R antagonist may be administered to a subject at a dose of about 0.0001 to about 10mg/kg of the subject's body weight, such as at a dose of about 1mg/kg to about 10mg/kg, at a dose of about 2mg/kg to about 9mg/kg, or at a dose of about 3mg/kg to about 8 mg/kg. In some embodiments, the IL-4R antagonist may be administered to the subject at a dose of about 1mg/kg, 2mg/kg, 3mg/kg, 4mg/kg, 5mg/kg, 6mg/kg, 7mg/kg, 8mg/kg, 9mg/kg, or 10 mg/kg.

In some embodiments, the methods disclosed herein comprise administering the IL-4R antagonist to the subject at a dosing frequency of about four times per week, twice per week, once per two weeks, once per three weeks, once per four weeks, once per five weeks, once per six weeks, once per eight weeks, once per twelve weeks, or less frequently, so long as a therapeutic response is achieved. In some embodiments, the methods disclosed herein comprise administering an IL-4R antagonist to a subject once every week, once every two weeks, once every three weeks, or once every four weeks. In some embodiments, the methods disclosed herein comprise administering an IL-4R antagonist to a subject once a month or twice a month.

In some embodiments, multiple doses of an IL-4R antagonist are administered to a subject over a prescribed period of time. In some embodiments, the methods of the present disclosure comprise sequentially administering multiple doses of an IL-4R antagonist to a subject. As used herein, "sequentially administering" refers to administering each dose of the IL-4R antagonist to a subject at different time points, e.g., on different days separated by a predetermined interval (e.g., hours, days, weeks, or months). In some embodiments, the methods of the present disclosure comprise sequentially administering a single initial dose of an IL-4R antagonist to a patient, followed by one or more second doses of the IL-4R antagonist, and optionally followed by one or more third doses of the IL-4R antagonist.

The terms "initial dose", "second dose" and "third dose" refer to the time sequence of administration of the IL-4R antagonist. Thus, an "initial dose" is the dose administered at the beginning of a treatment regimen (also referred to as a "loading dose"), a "second dose" is the dose administered after the initial dose, and a "third dose" is the dose administered after the second dose. The initial dose, the second dose, and the third dose may all contain the same amount of IL-4R antagonist, but may generally differ from one another in the frequency of administration. In certain embodiments, however, the amounts of IL-4R antagonist contained in the initial, second, and/or third doses are different from each other (e.g., adjusted upward or downward as appropriate) during the course of treatment. In certain embodiments, one or more (e.g., 1,2, 3, 4, or 5) doses are administered at the beginning of a treatment regimen as a "loading dose" followed by subsequent doses (e.g., a "maintenance dose") administered less frequently. In some embodiments, the initial dose or loading dose and the one or more second doses or maintenance doses each contain the same amount of the IL-4R antagonist. In other embodiments, the initial dose comprises a first amount of an IL-4R antagonist and the one or more second doses each comprise a second amount of an IL-4R antagonist. For example, the first amount of IL-4R antagonist may be 1.5x, 2x, 2.5x, 3x, 3.5x, 4x, or 5x or more of the second amount of IL-4R antagonist. In some embodiments, one or more maintenance doses of the IL-4R antagonist are administered in the absence of a loading dose.

In some embodiments, the loading dose is a "split dose" as two or more doses (e.g., 2, 3, 4, or 5 doses) administered on different days. In some embodiments, the loading dose is administered as a divided dose, wherein two or more doses are administered at least about one week apart. In some embodiments, the loading dose is administered as a divided dose, wherein two or more doses are administered about 1 week, 2 weeks, 3 weeks, or 4 weeks apart. In some embodiments, the loading dose is divided equally into two or more doses (e.g., half of the loading dose is administered as a first portion and half of the loading dose is administered as a second portion). In some embodiments, the loading dose is unevenly divided into two or more doses (e.g., more than half of the loading dose is administered as a first portion and less than half of the loading dose is administered as a second portion).

In some embodiments, each of the second and/or third doses is administered from 1 week to 14 weeks (e.g., ,1、1¹/₂、2、2¹/₂、3、3¹/₂、4、4¹/₂、5、5¹/₂、6、6¹/₂、7、7¹/₂、8、8¹/₂、9、9¹/₂、10、10¹/₂、11、11¹/₂、12、12¹/₂、13、13¹/₂、14、14¹/₂ or more) after the immediately preceding dose. As used herein, the phrase "immediately preceding dose" refers to a dose of an IL-4R antagonist being administered to a patient in a sequence of multiple administrations immediately prior to administration of the next dose in the sequence, without an intermediate dose.

The methods of the present disclosure can include administering any number of second and/or third doses of the IL-4R antagonist to the patient. For example, in certain embodiments, only a single second dose is administered to the patient. In other embodiments, the patient is administered the second dose two or more times (e.g., 2, 3, 4, 5, 6, 7, 8, or more times). Also, in certain embodiments, only a single third dose is administered to the patient. In other embodiments, the third dose is administered to the patient two or more times (e.g., 2, 3, 4, 5, 6, 7, 8, or more times).

In some embodiments involving a plurality of second doses, each second dose is administered at the same frequency as the other second doses. For example, each second dose may be administered to the patient 1 week, 2 weeks, 3 weeks, or 4 weeks after the immediately preceding dose. Similarly, in some embodiments involving multiple third doses, each third dose is administered at the same frequency as the other third doses. For example, each third dose may be administered to the patient 1 week, 2 weeks, 3 weeks, or 4 weeks after the immediately preceding dose. Alternatively, the frequency of the second dose and/or the third dose administered to the patient may vary during the course of the treatment regimen. The physician can also adjust the frequency of administration during the course of treatment, depending on the needs of the individual patient after clinical examination.

In some embodiments, a therapeutically effective amount of an IL-4R antagonist (e.g., an anti-IL-4R antibody) comprises 300mg administered every two weeks (Q2W). In some embodiments, a therapeutically effective amount of an IL-4R antagonist (e.g., an anti-IL-4R antibody) includes a loading dose of 600mg followed by one or more subsequent doses of 300mg administered every two weeks (Q2W). In some embodiments, no loading dose is administered.

In some embodiments, a therapeutically effective amount of an IL-4R antagonist (e.g., an anti-IL-4R antibody) comprises 200mg administered every two weeks (Q2W). In some embodiments, a therapeutically effective amount of an IL-4R antagonist (e.g., an anti-IL-4R antibody) includes a loading dose of 400mg followed by one or more subsequent doses of 200mg administered every two weeks (Q2W). In some embodiments, no loading dose is administered.

In some embodiments, a therapeutically effective amount of an IL-4R antagonist (e.g., an anti-IL-4R antibody) comprises 300mg administered every four weeks (Q4W). In some embodiments, a therapeutically effective amount of an IL-4R antagonist (e.g., an anti-IL-4R antibody) includes a loading dose of 600mg followed by one or more subsequent doses of 300mg administered every four weeks (Q4W). In some embodiments, a therapeutically effective amount of an IL-4R antagonist (e.g., an anti-IL-4R antibody) comprises a split loading dose of 600mg (e.g., wherein 300mg is administered on day 1 and 300mg is administered on day 15), followed by one or more subsequent doses of 300mg that begin Q4W administration four weeks after the day 15 administration. In some embodiments, no loading dose is administered.

In some embodiments, a therapeutically effective amount of an IL-4R antagonist (e.g., an anti-IL-4R antibody) comprises 200mg administered every four weeks (Q4W). In some embodiments, a therapeutically effective amount of an IL-4R antagonist (e.g., an anti-IL-4R antibody) includes a loading dose of 400mg followed by one or more subsequent doses of 200mg administered every four weeks (Q4W). In some embodiments, no loading dose is administered.

In some embodiments, for a subject having an age of No. 12 years to <18 years (e.g., a subject having moderate to severe or severe AD having an age of No. 12 years to <18 years), or for a subject having an age of No. 6 years to <18 years (e.g., a subject having moderate to severe or severe AD having an age of No. 6 years to <18 years), or for a subject having an age of No. 6 years to <12 years (e.g., a subject having moderate to severe or severe AD having an age of No. 6 years to <12 years), a therapeutically effective amount of an IL-4R antagonist (e.g., an anti-IL-4R antibody) comprises 300mg administered every two weeks (Q2W) if the subject weighs No. 60 kg. In some embodiments, if the subject weighs no less than 60kg, a loading dose of 600mg is administered to the subject followed by one or more subsequent doses of 300mg administered every two weeks (Q2W). In some embodiments, no loading dose is administered.

In some embodiments, for a subject having an age of ≡12 to <18 years (e.g., a subject having moderate to severe or severe AD having an age of ≡12 to <18 years), if the subject weighs <60kg, a therapeutically effective amount of an IL-4R antagonist (e.g., an anti-IL-4R antibody) comprises 200mg administered every two weeks (Q2W). In some embodiments, if the subject weighs <60kg, a loading dose of 400mg is administered to the subject followed by one or more subsequent doses of 200mg administered every two weeks (Q2W). In some embodiments, no loading dose is administered.

In some embodiments, for a subject having an age of No. 12 years to <18 years (e.g., a subject having moderate to severe or severe AD having an age of No. 12 years to <18 years), or for a subject having an age of No. 6 years to <18 years (e.g., a subject having moderate to severe or severe AD having an age of No. 6 years to <18 years), or for a subject having an age of No. 6 years to <12 years (e.g., a subject having moderate to severe or severe AD having an age of No. 6 years to <12 years), a therapeutically effective amount of an IL-4R antagonist (e.g., an anti-IL-4R antibody) comprises 200mg administered every two weeks (Q2W) if the subject weighs No. 30kg to <60 kg. In some embodiments, if the subject weighs greater than or equal to 30kg to <60kg, a 400mg loading dose is administered to the subject followed by one or more subsequent doses of 200mg administered every two weeks (Q2W). In some embodiments, no loading dose is administered.

In some embodiments, for a subject having an age of no less than 6 months to <6 years (e.g., a subject having moderate to severe or severe AD having an age of no less than 6 months to <6 years), or for a subject having an age of no less than 6 years to <12 years (e.g., a subject having moderate to severe or severe AD having an age of no less than 6 years to <12 years), or for a subject having an age of no less than 6 years to <18 years (e.g., a subject having moderate to severe or severe AD having an age of no less than 6 years to <18 years), the therapeutically effective amount of IL-4R antagonist (e.g., anti-IL-4R antibody) comprises 200mg administered every two weeks (Q2W) if the subject has a weight of no less than 15kg to <60 kg. In some embodiments, if the subject weighs no less than 15kg to <60kg, a loading dose of 400mg is administered to the subject followed by one or more subsequent doses of 200mg administered every two weeks (Q2W). In some embodiments, no loading dose is administered.

In some embodiments, for a subject having an age of no less than 6 months to <6 years (e.g., a subject having moderate to severe or severe AD having an age of no less than 6 months to <6 years), or for a subject having an age of no less than 6 years to <12 years (e.g., a subject having moderate to severe or severe AD having an age of no less than 6 years to <12 years), or for a subject having an age of no less than 6 years to <18 years (e.g., a subject having moderate to severe or severe AD having an age of no less than 6 years to <18 years), a therapeutically effective amount of an IL-4R antagonist (e.g., an anti-IL-4R antibody) comprises 300mg administered every four weeks (Q4W) if the subject has a weight of no less than 15kg to <60 kg. In some embodiments, if the subject weighs ∈15kg to <60kg, a loading dose of 600mg is administered to the subject followed by one or more subsequent doses of 300mg administered every four weeks (Q4W). In some embodiments, a split loading dose of 600mg (e.g., wherein 300mg is administered on day 1 and 300mg is administered on day 15) is administered to the subject followed by one or more subsequent doses of 300mg that begin Q4W administration four weeks after the day 15 administration. In some embodiments, no loading dose is administered.

In some embodiments, for a subject having an age of no less than 6 months to <6 years (e.g., a subject having moderate to severe or severe AD having an age of no less than 6 months to <6 years), or for a subject having an age of no less than 6 years to <12 years (e.g., a subject having moderate to severe or severe AD having an age of no less than 6 years to <12 years), or for a subject having an age of no less than 6 years to <18 years (e.g., a subject having moderate to severe or severe AD having an age of no less than 6 years to <18 years), the therapeutically effective amount of the IL-4R antagonist (e.g., anti-IL-4R antibody) comprises 300mg administered every four weeks (Q4W) if the subject has a weight of no less than 15kg to <30 kg. In some embodiments, if the subject weighs ∈15kg to <30kg, a loading dose of 600mg is administered to the subject followed by one or more subsequent doses of 300mg administered every four weeks (Q4W). In some embodiments, a split loading dose of 600mg (e.g., wherein 300mg is administered on day 1 and 300mg is administered on day 15) is administered to the subject followed by one or more subsequent doses of 300mg that begin Q4W administration four weeks after the day 15 administration. In some embodiments, no loading dose is administered.

In some embodiments, for a subject having an age of no less than 6 months to <6 years (e.g., a subject having moderate to severe or severe AD having an age of no less than 6 months to <6 years), or for a subject having an age of no less than 6 years to <12 years (e.g., a subject having moderate to severe or severe AD having an age of no less than 6 years to <12 years), or for a subject having an age of no less than 6 years to <18 years (e.g., a subject having moderate to severe or severe AD having an age of no less than 6 years to <18 years), the therapeutically effective amount of the IL-4R antagonist (e.g., anti-IL-4R antibody) comprises 200mg administered every four weeks (Q4W) if the subject has a weight of no less than 5kg to <15 kg. In some embodiments, if the subject weighs no less than 5kg to <15kg, a 400mg loading dose is administered to the subject followed by one or more subsequent doses of 200mg administered every four weeks (Q4W). In some embodiments, a split loading dose of 400mg (e.g., wherein 200mg is administered on day 1 and 200mg is administered on day 15) is administered to the subject followed by one or more subsequent doses of 200mg that begin Q4W administration four weeks after the day 15 administration. In some embodiments, no loading dose is administered.

Combination therapy

In some embodiments, the methods of the present disclosure comprise administering an IL-4R antagonist (e.g., an anti-IL-4R antibody) according to the present disclosure and one or more additional therapeutic agents to a subject (e.g., a pediatric or adolescent subject with a defect in bone growth). In some embodiments, the additional therapeutic agent is a topical therapeutic agent, e.g., TCS or a topical non-steroidal drug, such as TCI or clenbuterol. In some embodiments, the additional therapeutic agent is a systemic agent, such as cyclosporine a, methotrexate, mycophenolate mofetil (mycophenolate mofetil), azathioprine, systemic or oral corticosteroids, janus kinase (JAK) inhibitors, or interferon-gamma. In some embodiments, the additional therapeutic agent is an immune biologic, such as a tumor necrosis factor alpha (tnfa) inhibitor (e.g., an anti-tnfa antibody, such as infliximab), a CD11a inhibitor (e.g., an anti-CD 11a antibody, such as efalizumab), an IgE inhibitor (e.g., omalizumab) or a CD20 inhibitor (e.g., rituximab). As used herein, the expression "in combination with" means that the additional therapeutic agent is administered before, after, or simultaneously with the administration of the IL-4R inhibitor. The term "in combination with" also includes sequential or concomitant administration of an IL-4R inhibitor and an additional therapeutic agent.

For example, when administered "before" a pharmaceutical composition comprising an IL-4R antagonist, the additional therapeutic agent may be administered about 72 hours, about 60 hours, about 48 hours, about 36 hours, about 24 hours, about 12 hours, about 10 hours, about 8 hours, about 6 hours, about 4 hours, about 2 hours, about 1 hour, about 30 minutes, about 15 minutes, or about 10 minutes before administration of the pharmaceutical composition comprising the IL-4R antagonist. When administered "after" a pharmaceutical composition comprising an IL-4R antagonist, the additional therapeutic agent may be administered about 10 minutes, about 15 minutes, about 30 minutes, about 1 hour, about 2 hours, about 4 hours, about 6 hours, about 8 hours, about 10 hours, about 12 hours, about 24 hours, about 36 hours, about 48 hours, about 60 hours, or about 72 hours after administration of the pharmaceutical composition comprising the IL-4R antagonist. By "concurrently" or together with a pharmaceutical composition comprising an IL-4R antagonist is meant that the additional therapeutic agent is administered to the subject in a separate dosage form, or in a single combined dosage formulation comprising the additional therapeutic agent and the IL-4R antagonist, within less than about 10 minutes of (before, after, or simultaneously with) administration of the pharmaceutical composition comprising the IL-4R antagonist.

In some embodiments, the additional therapeutic agent is TCS. In some embodiments, the TCS is a moderately potent TCS. In some embodiments, the TCS is a low potency TCS. In some embodiments, the additional therapeutic agent is TCI. In some embodiments, the additional therapeutic agent is clenbuterol.

TABLE 1 informal sequence Listing

Examples

The following examples are put forth so as to provide those of ordinary skill in the art with a complete disclosure and description of how to make and use the methods and compositions of the present disclosure, and are not intended to limit the scope of what the inventors regard as their invention. Efforts have been made to ensure accuracy with respect to numbers used (e.g., amounts, temperature, etc.), but some experimental errors and deviations should be accounted for. Unless otherwise indicated, parts are parts by weight, molecular weight is average molecular weight, temperature is degrees celsius, and pressure is at or near atmospheric pressure.

EXAMPLE 1 Dupup Li Youshan anti-treatment of children with moderate to severe atopic dermatitis increased bone alkaline phosphatase (marker of bone mineralization)

The purpose of this analysis was to report the effect of the domino Li Youshan anti-treatment on markers of bone formation in children with moderate to severe AD aged no less than 6 to <12 years.

Method of

Serum from participants in LIBERTY AD PEDS (NCT 03345914) and LIBERTY AD PED-OLE (NCT 02612454) was analyzed retrospectively. In LIBERTY AD PEDS (a double-blind, 16-week, 3-phase trial), children aged 6 to <12 years were randomized 1:1:1 into 300mg degree of common Li Youshan antibody (300 mg q4 w) every 4 weeks, 2 weeks of weight-based degree of common Li Youshan antibody regimen (100 mg q2w for patients with a baseline weight of <30kg, and 200mg q2w for patients with a baseline weight of > 30 kg), or placebo, concomitant with moderate potency Topical Corticosteroids (TCS). After the first 16 weeks of trial, children aged 6 to <12 years were enrolled in open label extension study LIBERTY AD PED-OLE. Patient acceptance of Li Youshan against 300mg q4w, can be increased gradually in the event of inadequate clinical response at week 16 (200 mg q2w for patients with baseline body weight <60kg, and 300mg q2w for patients with baseline body weight > 60 kg), concomitant with moderate efficacy of TCS. Bone biomarkers (including BALP, procollagen type 1N-terminal propeptide, collagen type 1C-terminal cross-linked terminal peptide, osteocalcin, and insulin-like growth factor 1) were analyzed at baseline, 8 weeks, 12 weeks, 16 weeks, and only BALP at 52 weeks.

Results

Compared to patients in placebo group, at 16 weeks, the degree of common Li Youshan anti-treatment resulted in a rapid and significant increase in BALP geometric mean (standard error) levels in children with moderate to severe AD (77.7 (1.02) μg/L compared to 65.0 (1.04) μg/L; P < 0.0001). In addition, in children from the placebo group, a rapid and significant increase in BALP levels was observed once they were added to the OLE trial. The BALP levels in all treated children increased over 52 weeks to 78-84 μg/L, constituting a significant improvement over baseline, and comparable to the healthy reference interval. See fig. 1.

Both degree of common Li Youshan anti-dosing regimens resulted in a significant increase in the geometric mean (standard error) level of BALP at 8 weeks, 12 weeks and 16 weeks compared to placebo. For the 100/200mg q2w group, at week 8, BALP levels were 72.7 (1.03) μg/L for the day Li Youshan antibody and 62.0 (1.05) μg/L for the placebo group, P <0.0001, 74.7 (1.03) μg/L compared to 64.3 (1.05) μg/L at week 12, p=0.0002, and 78.0 (1.03) μg/L compared to 65.0 (1.04) μg/L, P <0.0001 at week 16. For 300mg q4w group, at week 8, BALP levels were 76.7 (1.03) μg/L for day Li Youshan and 62.0 (1.05) μg/L for placebo group, P <0.0001, at week 12 73.3 (1.04) μg/L compared to 64.3 (1.05) μg/L, p=0.002, at week 16 77.3 (1.03) μg/L compared to 65.0 (1.04) μg/L, P <0.0001. At week 52, BALP levels increased significantly from baseline (placebo vs. placebo converted to Dupu Li Youshan antibody: 64.2[1.04] μg/L vs. 82.9[1.04] μg/L, P <0.0001;100/200mg q2w:62.0[1.05] μg/L vs. 83.8[1.03] μg/L, P <0.0001;300mg q4w:64.1[1.04] μg/L vs. 78.7[1.04] μg/L, P < 0.0001), and also increased over the reference interval (Diemar et al, bone,2021, 146:115879).

An increasing trend was observed for other biomarkers (osteocalcin, PINP, IGF-1, and β -CTX) from baseline to 16-week-old Li Youshan anti-treatment, although the number of data points was limited due to insufficient serum volume available for analysis. See fig. 2-5. Overall, in this age group, the average biomarker levels measured in the dulp Li Youshan-resistant treated children improved from below the reference interval levels of osteocalcin, PINP and β -CTX to within the reference interval, and from below BALP and IGF-1 to near BALP and IGF-1.

Samples from girls and boys aged 6-12 years suffering from moderate to severe AD were analyzed by gender at BALP level in a subgroup, with the patient group analyzed at the beginning of the study aged 6-11 years. Although the reference intervals of BALP were different, girls exhibited higher values earlier and reached plateau around 12 years of age, while the level of BALP for boys continued to increase until around 15 years of age. (see Wu et al, ANN TRANSL MED,2021,9:40; lowe et al, J ALLERGY CLIN Immunol,2020,145:563-571;Silverberg,Pediatr Allergy Immunol.,2015,26:54-61; diemar et al, bone,2021, 146:115879). Treatment with dutch Li Youshan anti-treatment increased BALP levels in female and male patients to the reference interval and reflected this sex difference. At 16 weeks, the degree of common Li Youshan anti-treatment resulted in a rapid and significant increase in BALP geometric mean (standard error) levels for girls and boys (girls: 80.0 (1.04) μg/L versus 70.1 (1.06) μg/L, p=0.0018; boys: 75.7 (1.03) μg/L versus 60.4 (1.07) μg/L, P < 0.0001) compared to patients in the placebo group. Dupug Li Youshan anti-treatment resulted in an increased BALP level for all children receiving treatment, with girls reaching 90.5 μg/L and boys reaching 86.6 μg/L. See fig. 6-7.

A subgroup analysis was also performed to assess the effect of the domino Li Youshan anti-treatment on BALP levels in children aged 6-12 years with moderate to severe AD, with or without concomitant asthma. Regardless of asthma combination, the degree of common Li Youshan anti-treatment resulted in a rapid and significant increase in BALP geometric mean (standard error) levels at 16 weeks in children with moderate to severe AD compared to patients in placebo group (with asthma: 76.8[1.04] μg/L versus 59.1[1.07] μg/L, P <0.0001; without asthma: 78.5[1.03] μg/L versus 70.7[1.05] μg/L, p=0.0024). At week 52, geometric mean (standard error) BALP levels increased significantly compared to baseline and were comparable to the reference intervals for patients with and without asthma (placebo with asthma versus placebo converted to Dupu Li Youshan resistance: 62.3[1.06] μg/L versus 78.3[1.07] μg/L; dupu Li Youshan resistance: 62.1[1.04] μg/L versus 82.7[1.04] μg/L; placebo without asthma: placebo versus placebo converted to Dupu Li Youshan resistance: 66.0[1.06] μg/L versus 87.5[1.06] μg/L; dupu Li Youshan resistance: 64.0[1.04] μg/L versus 79.9[1.03] μg/L).

Conclusion:

the results of these placebo controls show for the first time that BALP increases rapidly and significantly and that other biomarkers may also increase in children with AD during treatment with the dutch Li Youshan antibody. These results indicate increased bone mineralization during treatment.

The scope of the invention is not limited by the specific embodiments described herein. Indeed, various modifications of the invention in addition to those described herein will become apparent to those skilled in the art from the foregoing description and accompanying drawings. Such modifications are intended to fall within the scope of the appended claims.

Claims (33)

1. A method for improving bone growth, comprising: selecting a subject having a bone growth defect, wherein the subject is a pediatric subject or an adolescent subject less than 18 years of age; and One or more doses of an interleukin-4 receptor (IL-4R) antagonist is administered to the subject, wherein the IL-4R antagonist is an anti-IL-4R antibody or an antigen-binding fragment thereof comprising three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3), wherein the HCDR1 comprises the amino acid sequence of SEQ ID NO:3, the HCDR2 comprises the amino acid sequence of SEQ ID NO:4, the HCDR3 comprises the amino acid sequence of SEQ ID NO:5, the LCDR1 comprises the amino acid sequence of SEQ ID NO:6, the LCDR2 comprises the amino acid sequence of LGS, and the LCDR3 comprises the amino acid sequence of SEQ ID NO:8. 2. The method of claim 1, wherein the subject suffers from atopic dermatitis (AD). 3. The method of claim 1 or 2, wherein the subject has moderate to severe or severe atopic dermatitis (AD). 4. The method of any one of claims 1 to 3, wherein the subject is a pediatric subject less than 12 years of age. 5. The method of claim 4, wherein the subject is between 6 and 11 years old. 6. The method of claim 4, wherein the subject is 6 months to 5 years old. 7. The method of any one of claims 1 to 3, wherein the subject is an adolescent subject between 12 and 17 years old. 8. The method of any one of claims 1 to 7, wherein the subject suffers from comorbid asthma. 9. The method of any one of claims 1 to 8, wherein the selecting step comprises selecting a subject who exhibits a level of a bone turnover marker below a threshold, wherein the bone turnover marker is bone-specific alkaline phosphatase, carboxyl-terminal cross-linking telopeptide of type I collagen (β-CTX), N-terminal propeptide of type I procollagen (PINP), insulin-like growth factor 1 (IGF-1), or osteocalcin. 10. The method of claim 9, wherein the threshold value is the average level of a bone turnover marker in a population of healthy subjects of the same age as the selected pediatric or adolescent subject. 11. The method of claim 9, wherein the bone turnover marker is bone-specific alkaline phosphatase. 12. The method of any one of claims 1 to 11, wherein the IL-4R antagonist is administered at a dose of about 50 mg to about 600 mg once a week (QW), once every two weeks (Q2W), once every three weeks (Q3W), or once every four weeks (Q4W). 13. The method of any one of claims 1 to 11, wherein the IL-4R antagonist is administered in an initial dose of 100-600 mg, followed by one or more subsequent doses of 50-300 mg, wherein each subsequent dose is administered one to four weeks after the immediately previous dose. 14. The method of any one of claims 1 to 5 and 7 to 13, wherein the subject is a pediatric subject between 6 and 11 years of age or an adolescent subject between 12 and 17 years of age, and wherein the subject has a baseline body weight of ≥ 60 kg, wherein the IL-4R antagonist is administered subcutaneously at an initial dose of 600 mg, followed by one or more subsequent doses of 300 mg Q2W. 15. The method of any one of claims 1 to 3 and 7 to 13, wherein the subject is an adolescent subject aged 12 to 17 years with a baseline weight of <60 kg, wherein the IL-4R antagonist is administered subcutaneously at an initial dose of 400 mg followed by one or more subsequent doses of 200 mg Q2W. 16. The method of any one of claims 1 to 5 and 8 to 13, wherein the subject is a pediatric subject 6 to 11 years of age with a baseline weight of ≥30 kg to <60 kg, wherein the IL-4R antagonist is administered subcutaneously at an initial dose of 400 mg followed by one or more subsequent doses of 200 mg Q2W. 17. The method of any one of claims 1 to 5 and 8 to 13, wherein the subject is a pediatric subject 6 to 11 years of age with a baseline weight of ≥15 kg to <30 kg, wherein the IL-4R antagonist is administered subcutaneously at an initial dose of 600 mg followed by one or more subsequent doses of 300 mg Q4W. 18. The method of any one of claims 1 to 5 and 8 to 13, wherein the subject is a pediatric subject 6 to 11 years of age with a baseline weight of ≥15 kg to <60 kg, wherein the IL-4R antagonist is administered subcutaneously at an initial dose of 300 mg on day 1, followed by an initial dose of 300 mg on day 15, followed by one or more subsequent doses of 300 mg Q4W starting four weeks after the day 15 dose. 19. The method of any one of claims 1 to 4, 6, and 8 to 12, wherein the subject is a pediatric subject 6 months to 5 years old with a baseline weight of ≥15 kg to <30 kg, wherein the IL-4R antagonist is administered subcutaneously at a dose of 300 mg Q4W. 20. The method of any one of claims 1 to 4, 6, and 8 to 12, wherein the subject is a pediatric subject 6 months to 5 years old with a baseline weight of ≥5 kg to <15 kg, wherein the IL-4R antagonist is administered subcutaneously at a dose of 200 mg Q4W. 21. The method of any one of claims 1 to 13 and 18 to 20, wherein the IL-4R antagonist is administered subcutaneously at an initial dose of 200 mg, followed by one or more subsequent doses of 200 mg, or at an initial dose of 300 mg, followed by one or more subsequent doses of 300 mg. 22. The method of any one of claims 1 to 21, wherein the IL-4R antagonist is administered in combination with a topical AD drug. 23. The method of claim 22, wherein the topical AD drug is a topical corticosteroid. 24. The method of any one of claims 1 to 23, wherein the IL-4R antagonist is administered for at least 16 weeks. 25. The method of any one of claims 1 to 24, wherein administration of the IL-4R antagonist for at least 16 weeks results in increased bone growth in the subject as measured by an increase in a bone turnover marker selected from the group consisting of bone-specific alkaline phosphatase, β-CTX, PINP, IGF-1, and osteocalcin. 26. The method of any one of claims 1 to 25, wherein the anti-IL-4R antibody or antigen-binding fragment thereof comprises a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO: 1 and a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO: 2. 27. The method of any one of claims 1 to 26, wherein the anti-IL-4R antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 9 and a light chain comprising the amino acid sequence of SEQ ID NO: 10. 28. The method of any one of claims 1 to 27, wherein the IL-4R antagonist is dupilumab. 29. The method of any one of claims 1 to 28, wherein the IL-4R antagonist is contained in a container selected from the group consisting of a glass vial, a syringe, a pre-filled syringe, a pen delivery device, and an autoinjector. 30. The method of claim 29, wherein the IL-4R antagonist is contained in a pre-filled syringe. 31. The method of claim 30, wherein the prefilled syringe is a single-dose prefilled syringe. 32. The method of claim 29, wherein the IL-4R antagonist is contained in an autoinjector. 33. The method of claim 29, wherein the IL-4R antagonist is contained in a pen delivery device.