Cc16-derived analogs for the treatment of inflammatory and obstructive lung diseases

Abstract

Described herein are compositions for treating inflammatory, neoplastic, and obstructive lung diseases characterized by CC16-derived analogs. The analogs can include N-terminal acetylation and C-terminal amidation, enhancing stability. Adopting a helical conformation, the CC16-derived analogs retain similar activity to full-length CC16 protein. The compositions are effective for a variety of lung diseases, allowing for nebulization, subcutaneous injection, and pulmonary delivery, with administration options including nasal inhalation, subcutaneous, or oral routes. A method for treating these diseases involves administering a CC16-derived analog to a subject in need thereof. The innovation provides a versatile, effective solution for respiratory diseases, providing a tailored approach to treatment.

Description

[0001] Cross-reference to related applications

[0002] This application claims the benefit of U.S. Provisional Application No. 63 / 611,467, filed December 18, 2023, the description of which is incorporated herein by reference in its entirety.

[0003] Statement regarding federally funded research or development

[0004] This invention was made with government funding under grant numbers HL142769 and AI135108 granted by the National Institutes of Health (NIH). The government holds certain rights in this invention.

[0005] Reference to the electronic sequence list

[0006] The contents of the electronic sequence list (ARIZ 2335 PCT.xml; size: 58,787 bytes; and creation date: December 17, 2024) are incorporated herein by reference in their entirety. Technical Field

[0007] The present invention is characterized by CC16-derived analogues for the treatment of inflammatory, neoplastic and obstructive lung diseases. Background Technology

[0008] CC16 (club cell secretory protein, also known as CCSP, CC10, and uterine globin) is a homodimeric lung protein encoded by the SCGB1A1 gene. Primarily produced by club cells and non-ciliated epithelial cells in the distal airways, CC16 diffuses into the bloodstream and is readily detectable in serum. Extensive research has been conducted to explore its potential as a biomarker of lung epithelial cell damage. Studies have shown reduced serum CC16 concentrations in patients with obstructive lung diseases such as asthma and chronic obstructive pulmonary disease (COPD) as well as in individuals with impaired lung function in the general population. In several epidemiological cohorts, low serum CC16 levels have been shown to predict impaired lung function development subsequently occurring in childhood, as well as accelerated decline in lung function and airflow limitation (a hallmark of COPD) in adulthood. Low serum CC16 levels also predict mortality from lung cancer.

[0009] There is an urgent need for adjunctive therapies for inflammatory, neoplastic, and obstructive lung diseases to effectively restore patients' CC16 levels. This invention addresses this need. Summary of the Invention

[0010] The object of this invention is to provide compositions and methods, as specified in the independent claims, that allow for the treatment of inflammatory, neoplastic, and obstructive lung diseases or the common cold (e.g., rhinovirus). Embodiments of the invention are given in the dependent claims. Embodiments of the invention may be freely combined with each other unless they contradict each other.

[0011] In some embodiments, the invention is characterized by a composition comprising a CC16-derived analog. In some embodiments, the CC16-derived analog comprises compound C836 according to the sequence DQD-Nle-REAGAQLKKLVDT (SEQ ID NO: 41). Therefore, in other embodiments, the invention may be characterized by a composition comprising a CC16-derived analog according to the sequence DQD-Nle-REAGAQLKKLVDT (SEQ ID NO: 41). In some implementations, CC16-derived analogues comprise sequences that are at least 100%, 95%, 90%, or 85% identical to the sequences according to SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, and SEQ ID NO: 41.

[0012] In some embodiments, the invention is characterized by a method of treating inflammatory, neoplastic, and obstructive pulmonary diseases in subjects with such need. In some embodiments, the method includes administering to the patient an effective amount of, for example, a CC16-derived analog as described herein. In other embodiments, the method includes administering to the patient an effective amount of a CC16-derived analog according to the sequence DQD-Nle-REAGAQLKKLVDT (SEQ ID NO: 41).

[0013] The CC16-derived analogs (e.g., peptides) described herein may further comprise N-terminal acetylation, C-terminal amidation, or a combination thereof. In some embodiments, the CC16-derived analogs are helical. In other embodiments, at least a portion of the CC16-derived analogs is cyclic.

[0014] In some embodiments, the CC16-derived analogues described herein retain activity similar to the full-length CC16 protein. In some embodiments, the composition is in a formulation for nebulization, subcutaneous injection, or pulmonary delivery. In some embodiments, the composition is administered via nasal inhalation, subcutaneous administration, or oral administration.

[0015] One of the unique and inventive technical features of this invention is the use of CC16-derived analogues (e.g., peptide mimics) for the treatment of inflammatory, neoplastic, and obstructive pulmonary diseases. While not intended to limit the invention to any particular theory or mechanism, the technical features of this invention are believed to offer a number of advantages over natural CC16. Specifically, the analogues are designed to deliver a higher dose of the active domain per gram of mass, thereby enhancing their therapeutic efficacy. Furthermore, the removal of oxidizable amino acids reduces the likelihood of loss of activity due to oxidation, thereby improving the stability of the analogues and extending their half-life. In addition, their smaller molecular size compared to CC16 facilitates more efficient delivery.

[0016] These combined features—smaller size, optimized active area, and enhanced stability—represent a significant improvement over CC16 and underscore the novelty and inventiveness of this invention. No existing references or work possesses the unique, inventive technical features of this invention.

[0017] Furthermore, existing references teach a departure from this invention. For example, current treatments for COPD and asthma primarily rely on inhaled corticosteroids, typically in combination with bronchodilators. In cases of acute exacerbations, systemic corticosteroids and antibiotics are commonly used. For asthma, biologics have emerged as a treatment option, but their use is typically limited to specific subgroups of severely ill patients due to their specific targets, high cost, and the need for injection. Treatment options for COPD are even more limited, with most relying on therapies originally developed for asthma. Severe cases may require lung transplantation or long-term oxygen therapy, significantly limiting patient independence and quality of life. The use of corticosteroids for both conditions is associated with numerous long-term side effects, highlighting the need for more effective and accessible treatments.

[0018] Furthermore, the inventive features of this invention lead to surprising results. For example, protein fragments containing less than 10% of the full-length CC16 protein can effectively replicate the functional properties of the native CC16 protein. Notably, some of these analogues are able to upregulate the expression of the endogenous CC16 gene and protein. Another surprising result of the inventive features of this invention is that, when CC16-derived analogues are delivered into the bloodstream, they affect lung function and host defense, more specifically by reducing airway hyperresponsiveness and inflammation, while promoting the secretion of protective factors, including antimicrobial agents, antiviral agents, and antioxidants.

[0019] Any feature or combination of features described herein is included within the scope of this invention, provided that, based on the context, this specification, and the knowledge of one skilled in the art, it can be determined that the features included in any such combination are not inconsistent with each other. Additional advantages and aspects of the invention will become apparent from the following detailed description and claims. Attached Figure Description

[0020] The features and advantages of the present invention will become apparent from the following detailed description presented in conjunction with the accompanying drawings, wherein:

[0021] Figure 1 Solid-phase synthesis of compound C836 is shown. (a) 10% piperidine in DMF, 2 + 20 min. (b) Coupling: (i) Fmoc-Thr(tBu), DIC / HOBt, (ii) Fmoc-Thr(tBu), HBTU / Calidin, in DMF, 2 h. (c) Universal solid-phase program. (d) 2.0 M acetic anhydride / pyridine in DCM, 20 min. (f) Universal TFA cleavage protocol: 95% TFA, 3% EDT, 2% TIPS, 2 h. (g) (i) Post-treatment and HPLC purification.

[0022] Figure 2 Solid-phase synthesis of cyclic peptide C966 is shown. (a) 10% piperidine in DMF, 2 + 20 min. (b) Coupling: (i) Fmoc-Thr(tBu), DIC / HOBt, (ii) Fmoc-Thr(tBu), HBTU / Calidin, in DMF, 2 h. (c) Universal solid-phase program. (d) 2.0 M acetic anhydride / pyridine in DCM, 20 min. (e) Pd(0)[(Ph3P)4], dimethylbarbituric acid, DCM, 2 x 30 min. (f) Cyclation of HBTU / DIEA in DMF, overnight. (g) Universal TFA cleavage protocol: 95% TFA, 3% EDT, 2% TIPS, 2 h. (h) Post-treatment and HPLC purification.

[0023] Figure 3The RP-HPLC chromatogram of compound C836 is shown. HPLC was performed under the following conditions: C836 was analyzed using an Agilent 2600 system with a 3.0 x 100 mm Agilent Poroshell 120 EC-C18 2.7 μm column, eluted with a solvent gradient A:B (where solvent A: 0.1% TFA in water and solvent B: 0.09% TFA in acetonitrile:water (9:1)) at a flow rate of 0.4 mL / min over 15 minutes. Rt = 7.419 min.

[0024] Figure 4 shows the application of CC16 - / - rCC16 in mice reduced human rhinovirus (RV) viral load. RT-PCR at 48 hours. *p<0.05 by t-test.

[0025] Figure 5A and Figure 5B The absence of CC16 was shown to lead to increased infectivity of mouse tracheal epithelial cells (MTECs) grown at the air-liquid interface (ALI). Figure 5A Shown after 48 hours at WT and CC16 - / - Mycoplasma pneumoniae (Mp) load in MTECS, and Figure 5B The RV loading is shown. Cells were grown at ALI for 2 weeks, after which they were fed with 1x10 8 Mp CFU / ml or 1x10 7 RV PFU / ml infection. Viral load was determined by RT-PCR using Taqman primers and is shown as a fold increase relative to WT. *p<0.05 and ****p<0.0001 were obtained by t-test.

[0026] Figure 6 The results showed that rCC16 and C836 reduced RV infection in human nasal cells. Cells were grown at ALI for 3 weeks and infected with RV for 4 hours, followed by washing and incubation for 48 hours with or without rCC16 or C836. RV was determined by qPCR. Each point represents one technical replicate, by ANOVA, **p<0.01.

[0027] Figure 7A and Figure 7B The study showed that nasal cells from asthmatic patients were more susceptible to RV infection compared to non-asthmatic participants. Figure 7A And it has reduced Scgb1a1 (CC16) gene expression. Figure 7B Cells were grown at ALI for 3 weeks and treated with RV (1x10⁻¹) 6PFU (Polydioxanone) was used to infect samples 4 hours later, followed by washing and incubation for 48 hours. RV and CC16 were identified by qPCR. Each spot represents one technical replicate, as per ANOVA, p < 0.0001.

[0028] Figure 8 This study demonstrates that C836 inhibits neutrophil recruitment and protects lung function. Left panel: C836 delivered via IV 2 hours after each HDM challenge. Cells in the lavage fluid were measured 24 hours after the last HDM challenge. Right panel: Baseline lung function in the absence of methacholine challenge, as assessed in mice receiving C836 IV, 24 hours after the last HDM challenge. By ANOVA, **p<0.01, ****p<0.0001.

[0029] Figure 9 C836 was shown to reduce AHR in the allergic airway. C836 was delivered to the airway 2 hours after each HDM challenge. AHR of acetylcholine was assessed 24 hours after the last HDM challenge. *p<0.05 at the highest dose.

[0030] Figure 10 The activity of peptide mimics in screening human nasal cells for reducing RV infection is shown. Top panel: RV infectivity; Middle panel: IFN-λ; Bottom panel: TNF-α level. All were determined by RT-PCR. Detailed Implementation

[0031] Various peptides, solvents, solutions, carriers, and / or components to be used in the preparation of compositions to be used in the methods disclosed herein are disclosed. Various steps, elements, amounts, routes of administration, symptoms and / or treatments used or observed when performing the disclosed methods are also disclosed, as well as the methods themselves. These and other materials, steps, and / or elements are disclosed herein, and it should be understood that while specific references to every individual and collective combination and arrangement of these materials may not be explicitly disclosed when disclosing combinations, subsets, interactions, groups, etc., each is specifically contemplated and described herein. It should also be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting.

[0032] Unless otherwise explained, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the disclosed invention pertains. The singular terms “a,” “an,” and “the” include plural referents unless the context clearly indicates otherwise. Similarly, the word “or” is intended to include “and” unless the context clearly indicates otherwise. The term “comprising” means that other elements may be present in addition to the defined elements presented. The use of “comprising” indicates inclusion rather than limitation. In other words, the term “comprising” means “primarily includes, but not necessarily uniquely includes.” Furthermore, variations of the word “comprising” (such as “comprise” and “comprises”) have the corresponding equivalent meanings. In one aspect, the techniques described herein relate to the compositions, methods, and corresponding components described herein, which are essential to the invention but open to including unspecified elements (necessary or non-essential) (“comprising”).

[0033] All publications, patent applications, patents and other references mentioned herein are incorporated herein by reference in their entirety for all purposes. In case of conflict, this specification (including the interpretation of terminology) shall prevail.

[0034] While methods and materials similar to or equivalent to those described herein may be used to practice or test the disclosed techniques, suitable methods and materials are described below. The materials, methods, and examples are illustrative only and are not intended to be limiting.

[0035] The terms “polypeptide” and “protein” are used interchangeably to refer to a polymer of amino acid residues, containing both native and non-native amino acid residues, and are not limited to a minimum length. Therefore, peptides, oligopeptides, dimers, polymers, etc., are included in this definition. Both full-length proteins and fragments thereof are covered in this definition. The term also includes post-translational modifications of polypeptides, including, for example, glycosylation, sialylation, acetylation, and phosphorylation. Furthermore, “polypeptide” as used herein also refers to modified proteins, such as the deletion, addition, and substitution of single or multiple amino acid residues in the native sequence, provided that the protein retains the desired activity. For example, serine residues may be substituted to eliminate a single reactive cysteine ​​or remove a disulfide bond, or conserved amino acid substitutions may be performed to eliminate cleavage sites. These modifications can be intentional, such as through site-directed mutagenesis, or can be accidental, such as through mutations in the host producing the protein or due to errors in polymerase chain reaction (PCR) amplification.

[0036] As used herein, the term "peptide" refers to a short polymer of amino acids linked together by peptide bonds. Peptides are approximately 50 amino acids or less in length compared to other amino acid polymers, such as proteins, polypeptides, etc. Peptides can contain natural amino acids, non-natural amino acids, amino acid analogs, and / or modified amino acids. Peptides can be subsequences of naturally occurring proteins or non-natural (synthetic) sequences.

[0037] In some embodiments, the peptides described herein are modified by adding an amine or acid group to the C-terminus and acetylation or adding a histidine (H) to the N-terminus. In other embodiments, the peptides described herein are modified by adding an acid to the N-terminus. Non-limiting examples of acids that can be used to modify the N-terminus of the peptides described herein may include, but are not limited to, hydroxyl groups (-OH), carboxyl / carboxylic acid groups (COOH), or combinations thereof. In further embodiments, the peptides described herein are modified with lipid groups and with the C-terminus and / or N-terminus.

[0038] As used herein, the term "wildtype" refers to a non-mutated version of a gene, allele, genotype, polypeptide, or phenotype, or a fragment of any of them. It can exist in nature or be generated through recombination. As used herein, the term "variant" refers to a nucleic acid molecule or polypeptide that differs from a reference nucleic acid molecule or polypeptide by substitution, deletion, and / or addition of one or more amino acids and substantially retains at least one biological activity of the reference nucleic acid molecule or polypeptide.

[0039] The term "peptide mimetic" or "peptidomimetic" refers to a peptide-like molecule that mimics a sequence derived from a protein or peptide. Peptide mimetic or pepttidomimetic molecules can contain amino acid and / or non-amino acid components. Examples of peptide mimetic molecules include chemically modified peptides, peptide-like molecules (with side chains attached to the nitrogen atom of the peptide backbone instead of the α-carbon), P-peptides (with amino groups bonded to the β-carbon instead of the α-carbon), and so on. As used herein, "peptide mimetic," "peptide analog," and "analyte" are used interchangeably.

[0040] As used herein, "conservative" amino acid substitution refers to the substitution of an amino acid in a peptide or polypeptide with another amino acid having similar chemical properties, such as size or charge. For the purposes of this disclosure, each of the following eight groups contains amino acids that are conservedly substituted for each other:

[0041] 1) Alanine (A) and glycine (G);

[0042] 2) Aspartic acid (D) and glutamic acid (E);

[0043] 3) Asparagine (N) and glutamine (Q);

[0044] 4) Arginine (R) and lysine (K);

[0045] 5) Isoleucine (I), leucine (L), methionine (M), and valine (V);

[0046] 6) Phenylalanine (F), tyrosine (Y), and tryptophan (W);

[0047] 7) Serine (S) and threonine (T); and

[0048] 8) Cysteine ​​(C) and methionine (M).

[0049] Naturally occurring residues can be categorized based on common side-chain properties, such as: polar positive (histidine (H), lysine (K), and arginine (R)); polar negative (aspartic acid (D), glutamic acid (E)); polar neutral (serine (S), threonine (T), asparagine (N), glutamine (Q)); nonpolar aliphatic (alanine (A), valine (V), leucine (L), isoleucine (I), methionine (M)); nonpolar aromatic (phenylalanine (F), tyrosine (Y), tryptophan (W)); proline and glycine; cysteine. As used herein, "semi-conservative" amino acid substitution refers to the substitution of an amino acid in a peptide or polypeptide with another amino acid from the same category.

[0050] In some embodiments, unless otherwise stated, conserved or semi-conserved amino acid substitutions may also cover non-naturally occurring amino acid residues having chemical properties similar to native residues. These non-natural residues are typically incorporated through chemical peptide synthesis rather than through synthesis in biological systems. These include, but are not limited to, peptide mimics and other reversed or inverted forms of the amino acid moiety. In some embodiments, the embodiments described herein may be limited to natural amino acids, non-natural amino acids, and / or amino acid analogs.

[0051] Non-conservative substitution can involve exchanging a member of one category for a member from another category.

[0052] The terms “subject,” “individual,” “host,” “animal,” and “patient” are used interchangeably in this article to refer to mammals, including but not limited to rodents, monkeys, humans, felines, canines, equines, bovines, swine, sheep, capillaries, laboratory mammals, farm mammals, sporting mammals, and pet mammals.

[0053] As used herein, the terms “administration” and “administering” refer to the act of giving a drug, prodrug, or other agent or therapeutic treatment (e.g., SP-A peptide) to a subject or to cells, tissues, and organs in vivo, in vitro, or ex vivo. Exemplary routes of administration to the human body may include the subarachnoid space of the brain or spinal cord (intrathecal), eye (ocular), mouth (oral), skin (local or percutaneous), nose (nasal cavity), lungs (inhalation), oral mucosa (buccal), ear, rectum, vagina, and by injection (e.g., intravenous, subcutaneous, intratumoral, intraperitoneal, etc.).

[0054] As used herein, “treatment” encompasses any administration or application of a therapeutic agent for diseases in mammals, including humans, and includes suppressing, preventing or alleviating disease, such as by inducing remission, or restoring or repairing lost, missing or defective functions; or stimulating inefficient processes.

[0055] "Pharmaceutically acceptable carrier" refers to a non-toxic solid, semi-solid, or liquid filler, diluent, encapsulating material, formulation adjuvant, or carrier conventional in the art, used in conjunction with a therapeutic agent intended for administration to a subject. A pharmaceutically acceptable carrier is non-toxic to the recipient at the dose and concentration used and is compatible with other components of the formulation. A pharmaceutically acceptable carrier is suitable for the formulation in which it is used. For example, if the therapeutic agent is to be administered orally, the carrier may be a gelatin capsule. If the therapeutic agent is to be administered subcutaneously, the carrier ideally is non-irritating to the skin and does not cause an injection site reaction.

[0056] Now for reference Figures 1 to 8 The present invention is characterized by CC16-derived peptide mimics (i.e., analogs) for the treatment of inflammatory, neoplastic, and obstructive pulmonary diseases. CC16-derived peptide mimics offer several advantages over conventional proteins, making them highly attractive for therapeutic development. Their resistance to proteolytic and oxidative degradation improves metabolic stability and prolongs in vivo half-life, enhancing pharmacokinetics. Higher stability also improves shelf life, reduces denaturation, and minimizes degradation during storage. Structural modifications allow for enhanced activity, while precise design reduces off-target effects. Incorporation of non-natural building blocks (amino acids) reduces immunogenicity, as peptide mimics are less likely to be recognized and targeted by the immune system, and they are synthesized to minimize biocontamination. Their production relies on direct chemical synthesis rather than complex expression systems, providing a cost-effective manufacturing process. Furthermore, peptide mimic modifications can improve membrane permeability, facilitate intracellular delivery, and allow for fine-tuning of solubility to ensure compatibility with various routes of administration through customized hydrophobic or hydrophilic properties.

[0057] In some embodiments, the invention is characterized by a composition comprising a CC16-derived analog. For example, the composition comprises a CC16-derived analog according to the sequence: Ac-Asp-Gln-Asp-Nle-Arg-Glu-Ala-Gly-Ala-Gln-Leu-Lys-Lys-Leu-Val-Asp-Thr-NH2. Therefore, in other embodiments, the invention may be characterized by a composition comprising a CC16-derived analog according to the sequence DQD-Nle-REAGAQLKKLVDT (SEQ ID NO: 41). In some implementations, CC16-derived analogues comprise sequences that are at least 100%, 95%, 90%, or 85% identical to the sequences according to SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, and SEQ ID NO: 41.

[0058] Non-limiting examples of analogues derived from CC16 may include, but are not limited to, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40 and SEQ ID NO: 41.

[0059] In some embodiments, the CC16-derived analogs (e.g., peptides) described herein may further comprise N-terminal acetylation, C-terminal amidation, or a combination thereof. The CC16-derived analogs are helical and retain activity similar to the full-length CC16 protein. Therefore, in some embodiments, the CC16-derived analogs are helical. In some embodiments, the CC16-derived analogs are cyclic. In other embodiments, at least a portion of the CC16-derived analog is cyclic.

[0060] In some embodiments, the composition is used to treat inflammatory, neoplastic, and obstructive lung diseases. In some embodiments, the composition is in a formulation for nebulization, subcutaneous injection, or pulmonary delivery. In some embodiments, the composition is administered via nasal inhalation, subcutaneous administration, or oral administration.

[0061] In other embodiments, the invention is characterized by a method of treating inflammatory, neoplastic, and obstructive pulmonary diseases in a subject with such need. The method includes administering to the patient an effective amount of a CC16-derived analog as described herein. In other embodiments, the method includes administering to the patient an effective amount of a CC16-derived analog according to the sequence DQD-Nle-REAGAQLKKLVDT (SEQ ID NO: 41).

[0062] Non-limiting examples of inflammatory, neoplastic, and obstructive lung diseases include, but are not limited to, asthma, chronic obstructive pulmonary disease (COPD), cystic fibrosis, lung cancer, impaired lung function, and the conditions that cause impaired obstructive lung function.

[0063] As used in this article, "obstructive pulmonary function deficit" refers to a condition that restricts airflow, preventing the lungs from effectively taking in oxygen or expelling carbon dioxide. These deficits typically result in shortness of breath and reduced vital capacity, and are usually diagnosed through pulmonary function tests, with spirometry being a commonly used method. Obstructive pulmonary function deficit is characterized by a reduced level of forced expiratory volume in one second (FEV1) and the ratio of the reduction in FEV1 to forced vital capacity (FEV1 / FVC). Examples of conditions that can lead to obstructive pulmonary function deficit include preterm birth, which is associated with lower blood levels of CC16, potentially leading to airflow limitation later in life.

[0064] Obstructive lung function deficits can have both genetic and environmental causes. Some individuals are born with reduced lung function, while others who initially have normal lung function may experience a decline due to events such as smoking or pneumonia. In some implementations, both asthma and COPD are considered obstructive lung diseases.

[0065] The invention may also be characterized in a method of using a CC16-derived analog composition for treating inflammatory, neoplastic, and obstructive pulmonary diseases in subjects with such need. In other embodiments, the invention may also be characterized in a method of using a CC16-derived analog composition for treating inflammatory, neoplastic, and obstructive pulmonary diseases in subjects with such need; wherein the CC16-derived analog composition comprises the sequence according to SEQ ID NO: 41.

[0066] Example 1

[0067] The following are non-limiting embodiments of the present invention. It should be understood that these embodiments are not intended to limit the invention in any way. Equivalents or alternatives are within the scope of the present invention.

[0068] Non-standard abbreviations:

[0069] Aloc allyloxycarbonyl

[0070] Allyl

[0071] Boc tert-butoxycarbonyl

[0072] DCM dichloromethane

[0073] DIC N,N'-Diisopropylcarbodiimide

[0074] DIEA (Diisopropylethylamine)

[0075] DMF N,N'-dimethylformamide

[0076] DMSO (dimethyl sulfoxide)

[0077] EDT 1,2-ethylenedithiol

[0078] ESI-MS Electrospray Ionization-Mass Spectrometry

[0079] EtOAc (ethyl acetate)

[0080] Et2O diethyl ether

[0081] Fmoc (9H-fluoren-9-ylmethoxy)carbonyl

[0082] HBTU 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethylurea hexafluorophosphate

[0083] HOBt N-hydroxybenzotriazole

[0084] HPLC (High Performance Liquid Chromatography)

[0085] OBt O-benzotriazolyl

[0086] MeOH (methanol)

[0087] NMR (Nuclear Magnetic Resonance)

[0088] Pbf 2,2,4,6,7-pentamethyl-dihydrobenzofuran-5-sulfonyl

[0089] RP-HPLC (Reversed-Phase High-Performance Liquid Chromatography)

[0090] SPE solid phase extraction

[0091] SPPS solid-phase peptide synthesis

[0092] tBu tert-butyl

[0093] THF Tetrahydrofuran

[0094] TFA (trifluoroacetic acid)

[0095] TIPS: Triisopropylsilane

[0096] TMS Tetramethylsilane

[0097] TRT Triphenylmethyl

[0098] Synthesis of CC16 compound analogs:

[0099] Chemical materialsN-α-Fmoc-protected amino acids, HBTU, DIC, Oxima, and HOBt were purchased from P3 Biosystems (Louisville, KY) or Novabiochem (San Diego, CA). Rink and Wang resins were obtained from Rapp Polymere (Tubingen, Germany) or Novabiochem (San Diego, CA). For Nα-Fmoc-protected amino acids, the following side-chain protecting group was used: Fmoc-Arg(N w Fmoc-Pbf), Fmoc-Asp(OtBu), Fmoc-Asp(OAll), Fmoc-Gln(Nc-Trt), Fmoc-Glu(OtBu), Fmoc-Glu(OAll), Fmoc-Lys(Ne-Boc), Fmoc-Lys(Ne-Aloc), Fmoc-Thr(tBu). Reagent-grade solvents, reagents, and acetonitrile used for HPLC were obtained from VWR (West Chester, PA) or Aldrich-Sigma (Milwaukee, WI) and used without further purification unless otherwise specified. Chemicals and reagents were obtained from Sigma-Aldrich or TCI. Solid-phase synthesis was performed using a Domino manual synthesizer from Torviq (Tucson, AZ) in a sintered syringe.

[0100] Universal synthesis All solution-phase reactions were carried out in oven-dried glassware under an argon (Ar) atmosphere. All chemicals were obtained from commercial sources and used without further purification. 1 H NMR spectra were recorded on a Bruker-DRX-300 MHz instrument, with chemical shifts reported relative to TMS (0.0 ppm) and residual DMSO (2.50 ppm). Proton decoupling... 13 C10 NMR spectra were referenced to CDCl3 (77.0 ppm) and DMSO (39.51 ppm). Low-resolution mass spectra were obtained using AP-ESI on an AGILENT (HP) MDS 1100. High-resolution mass spectra (HRMS) were recorded on a JEOL HX110A instrument.

[0101] Universal solid-phase peptide synthesis :like Figure 1As outlined in the previously published Fmoc / tBu synthetic strategy and standard HOBt / DIC, HBTU, or symmetric anhydride activation, peptides were prepared via solid-phase synthesis on Rink amide polystyrene (PS) resin (0.68 mmol / g) or Wang PS resin (0.54 mmol / g). 𝛼 -Fmoc amino acids are bicoupled using pre-activated 0.3 M HOBt ester and HBTU / 2,4,6-coridine or symmetrical anhydride coupling.

[0102] The Rink resin was washed with DMF, and nitrogen was removed with 1:10 piperidine in the DMF. 𝛼 -Fmoc protecting group (once for 2 minutes and once for 20 minutes). The resin was washed sequentially with DMF, DCM, DMF, bromophenol blue in 0.2 M HOBt, 0.05 mM solution in DMF, and then washed with DMF. N 𝛼 -Fmoc amino acids are coupled using a pre-activated 0.3 M HOBt ester in a DMF-DCM mixture (3 equivalents of acid, 3 equivalents of HOBt, and 3 equivalents of DIC). The resin slurry is stirred for 2 hours or until the bromophenol blue test becomes negative. If the test fails, the resin is washed with DMF, and the amino acid is recoupled for 3 hours using an HBTU / 2,4,6-dimethylpyridine procedure (3 equivalents of acid, 3 equivalents of HBTU, and 6 equivalents of 2,4,6-dimethylpyridine in a 0.3 M solution in DMF), or by using a pre-formed symmetrical anhydride (3 equivalents of acid and 3 equivalents of DIC in a 1:1 DMF-THF mixture) until the Kaiser test is negative. If coupling does not result in a negative Kaiser test, the resin is washed with DMF and the free amino groups are capped with 1.0 M acetic anhydride / pyridine in DMF for 20 minutes. After all the resins were sequentially coupled to the Rink amide resin, the resin was capped with 1.0 M acetic anhydride / pyridine in DMF for 20 min, washed with DMF, washed with DCM, dried under vacuum, and then stored in a refrigerator. To cleave the final product, the resin was thoroughly washed with DMF (3x) and DCM (7x). A cleavage mixture (10 mL / 1 g resin) containing CF3CO2H (95%), EDT (3%), and TIPS (2%) or a mixture thereof was injected into the resin, and the mixture was stirred at room temperature for 2 to 4 h. The solution was filtered, the resin was washed with CF3CO2H (2 x 3 min), the liquid phase was collected and concentrated under a nitrogen stream, and the product was precipitated using cold Et2O. The crude product was washed three times with cold Et2O, lyophilized, purified, and characterized as described above. The pure compound was dissolved in deionized (DI) or DMSO at a concentration of approximately 1 to 5 mM, and the concentration was determined by Trp-HPLC.

[0103] General solutions for cyclic peptides :like Figure 2 As outlined in the general synthetic protocol for compound C966, cyclic peptides are prepared using Rink amide PS resin (0.68 mmol / g). Standard Fmoc-protected amino acids are used in the synthesis, in addition to the Aloc-protected lysine building blocks Fmoc-Lys(Aloc)-OH and O-allyl ester-protected aspartic acid Fmoc-Asp(OAll) or O-allyl ester-protected glutamic acid Fmoc-Glu(OAll) used to incorporate orthogonally protected Lys / Asp or Lys / Glu pairs at the desired cyclization sites; Fmoc-Lys(Aloc)-OH, Fmoc-Asp(OAll)-OH, or Fmoc-Glu(OAll)-OH are coupled at the appropriate sites. Typically, 3 equivalents of Fmoc-protected amino acids, 3 equivalents of HCTU, and 3 equivalents of DIPEA are used in the coupling cycle. Fmoc deprotection was achieved by treatment with 10% piperidine / DMF for 2 + 20 min, except that the Fmoc deprotection time was reduced to 2 + 7 min after incorporation of allyl ester. For N-terminal acetylated peptides, the N-terminus was acetylated with 1.0 M acetic anhydride / pyridine in DMF for 20 min. For Aloc / allyl deprotection, the resin was washed with DCM and then purged with argon for 3 to 10 min. A pyrolysis mixture of 5 equivalents of dimethyl barbituric acid and 0.2 equivalents of Pd(0)[(Ph3P)4] in DCM (0.1 equivalent) was purged with argon and injected. The reaction mixture was stirred for 30 min and then repeated. The resin was washed sequentially with DCM, 10% DIEA in DCM (3 x 2 min), DMF (2 x), 1% sodium diethyldithiocarbamate trihydrate in DMF (2 x 20 min), 10% DIEA in DMF (3 x 2 min), and DMF. For cyclization, a solution of PyBOP (4 equivalents, 0.5 M) and DIPEA (8 equivalents) in DMF was added to the resin. Complete conversion of the side-chain crosslinked lactam was monitored by pyrolysis of a small sample followed by HPLC (typically about 6 hours). After completion, the resin was washed with DMF and DCM. To pyrolyze the final product, a pyrolysis mixture (10 mL / 1 g resin) consisting of CF3CO2H (95%), EDT (3%), and TIPS (2%) was injected into the resin, and the mixture was stirred at room temperature for 2 to 4 hours. The crude product was obtained using a general solid-phase procedure.

[0104] General QC and purification proceduresProduct purity was assessed using analytical RP-HPLC on an Agilent 2600 system with a 3.0 x 100 mm Agilent Poroshell 120 EC-C18 2.7 μm column. Elution was performed using a solvent gradient A:B (solvent A: 0.1% TFA in water, and solvent B: 0.09% TFA in acetonitrile:water (9:1)) over a period of 15 min at a flow rate of 0.4 mL / min. Ligand purification was performed on a Waters 600 HPLC system using a reversed-phase column (Vydac C18, 15–20 µm, 22 x 250 mm). Peptides were eluted with a linear gradient of CH3CN / 0.1% CF3CO2H at a flow rate of 5.0 mL / min. Separation was monitored at 230 nm and 280 nm. Size exclusion chromatography was performed on a borosilicate glass column (2.6 x 250 mm, Sigma, St. Louis, Missouri, MO) packed with medium-sized Sephadex G-25 or G-10 columns. The compounds were eluted isocratically with 1.0 M aqueous acetic acid. The pure compounds were dissolved in deionized water or DMSO at concentrations of approximately 1 to 5 mM. Accurate concentrations were determined by HPLC at 280 nm. Accordingly, a solution of D-Trp in water or DMSO was co-injected as an internal standard. The product C836 was 56 mg as a white lyophilized product (yield 21%, against (M+2H)2). + The calculated value was 971.04, HPLC >99%, see [reference]. Figure 3 Yields and retention times are depicted in Tables 1 through 3.

[0105] Table 1: Bold and underlined amino acids (i.e., Ala) indicate specific alanine residues used for activity scanning.

[0106]

[0107]

[0108] Table 2: Bold and underlined amino acids indicate which amino acids in the compound have changed compared to the wild-type CC16 protein.

[0109]

[0110] Table 3: Bold and underlined amino acids indicate which amino acids in the compound have changed compared to the wild-type CC16 protein. The "c" outside parentheses indicates which amino acids in the analogue have been cyclized.

[0111]

[0112] Example 2

[0113] The following are non-limiting embodiments of the present invention. It should be understood that these embodiments are not intended to limit the invention in any way. Equivalents or alternatives are within the scope of the present invention.

[0114] In the embodiments provided below, recombinant human CC16 (rCC16) is delivered directly to the lungs via retroorbital intravenous (IV) injection or oropharyngeal delivery. Both delivery methods effectively provide protection against airway hyperresponsiveness and respiratory infections. However, intravenous delivery alone has been shown to reduce leukocyte recruitment to the lungs.

[0115] rCC16 treatment reduces CC16 - / - Human rhinovirus (RV; RV1B) load in mice: For these studies, female mice were administered RV (4 x 10⁻⁶) via intranasal instillation. 7 Mice were infected with plaque-forming units (PFU) / 50 µl for 48 hours, and were given either a vector (saline) or rCC16 (human recombinant CC16) at infection time. RV load was determined by RT-PCR, and mice receiving rCC16 showed a significant reduction in RV compared to mice treated with the vector (Figure 4). This suggests that the host's response to the virus can be restored when CC16 is returned to the lungs.

[0116] Epithelial cells lacking CC16 exhibit enhanced infectivity. Compared to WT cells, CC16... - / - Mouse tracheal epithelial cells (MTEC) showed enhanced Mycoplasma pneumoniae (Mp) infection. Figure 5A To determine whether the role of CC16 during viral infection is epithelial-driven, rather than through affecting local immune cells, samples were collected from WT and CC16 at the air-liquid interface (ALI). - / - MTEC cultures were grown in mice and challenged with RV1B (human rhinovirus 1B). For these studies, MTEC infection was performed for 4 hours, followed by Transwell washing to remove any non-adherent viruses. Cells were then allowed to incubate at 35°C for 48 hours, after which RT-PCR was performed for RV infectivity. Figure 5B As shown, compared with WT cells, cells lacking CC16 exhibited significantly enhanced RV infection after 48 hours, indicating impairment in the antiviral host response and impairment in the antimicrobial response observed with Mp. Figure 5A ).

[0117] rCC16 and CC16-derived peptide mimics reduced RV infection in human nasal cells. To test whether exogenously added rCC16 could protect against RV virus infection, primary human nasal epithelial cells (HNECs) from non-asthmatic participants were grown at the air-liquid interface (ALI) for 21 days. On the day of infection, RV was added to the top surface and allowed to incubate for 4 hours, after which the cells were gently washed with culture medium and then added separately to rCC16 (25 g / ml) or the CC16-derived peptide mimic C836 (25 g / ml), and allowed to incubate for another 48 hours. RV infectivity was determined by PCR after harvest. Figure 6 As shown, both rCC16 and C836 significantly reduced RV infection in human nasal cells by approximately 80%.

[0118] Nasal cells from asthma patients exhibit reduced Scgb1a1 (CC16) gene expression and are more susceptible to RV infection. Airway cells isolated from asthma patients are also more susceptible to RV infection. Figure 7A Therefore, the enhanced susceptibility of RV in asthma patient cells may be due to reduced CC16 (also in...). Figure 7B (As shown in the figure) and thus lead to a decrease in CC16-dependent epithelial antiviral response protein.

[0119] Administration of the circulating CC16-derived peptide mimic C836 has a protective effect in house dust mite (HDM) allergic airway disease. Upregulation of CC16 during respiratory provocation has a protective effect through several approaches, including adenovirus overexpression, vitamin A-induced upregulation, and direct delivery of human recombinant CC16 (rCC16) and vesicle-encapsulated nanoparticles. While these strategies for increasing CC16 levels in the airway space have been successful, their clinical translation has been fraught with various complications, including delivery to patients. For example, adenovirus drug delivery is often unsuccessful due to limited efficacy and safety concerns. Similarly, the use of vitamin A as a therapeutic agent is problematic because excessive doses can lead to serious side effects, including an increased risk of cancer. Nanoparticle-based approaches, while innovative, are still in their early stages, with methods for their formulation and delivery not yet clearly defined in the literature, posing an additional obstacle to their practical application.

[0120] Therefore, small peptide mimics were designed based on the newly discovered LVD (leucine-valine-aspartic acid) integrin-binding domain contained within the CC16 protein. The lead compound C836 is a 17-mer containing the LVD integrin-binding domain. Figure 8Preliminary data (left) show that fewer neutrophils were recruited to the lungs during HDM provocation when C836 was administered intravenously (IV). The presence of neutrophils in the lungs of individuals with asthma is typically associated with a more severe and difficult-to-treat form of the condition. In COPD, neutrophils are often highly destructive, releasing factors and enzymes that degrade the basement membrane and damage fragile airway epithelium. Therefore, strategies to reduce neutrophil accumulation or activity in both asthma and COPD would provide significant benefits to overall lung health, potentially improving disease outcomes and reducing airway damage.

[0121] Furthermore, lung function tests under unprovocation conditions showed that C836 administration to HDM mice prevented lung function loss, as measured by FEV0.2 in an invasive PFT test. Figure 8 (Right). Lung function tests are typically performed using methacholine provocation, which induces bronchoconstriction. However, the tests mentioned here were performed without provocation, meaning that when C836 was administered along with HDM provocation, it prevented the loss of lung function typically associated with HDM exposure. These tests were performed at baseline without the use of bronchodilators, similar to a normal day in an asthma patient without an attack.

[0122] C836 reduced the allergic airway risk (AHR) in Cc16- / - mice exposed to HDM. The activity of C836 was tested to determine its ability to provide protection in an HDM-induced allergic airway model. CC16 was used for this purpose. - / - Mice were administered C836 (or a mediator control) via the oropharyngeal method approximately 1 hour after each HDM dose on days 0, 7, and 14. Mice were evaluated 24 hours after the last HDM challenge using flviVent® and methacholine challenge. Figure 9 As shown, C836 resulted in significantly lower airway hyperresponsiveness (AHR) compared to HDM treatment with the administered medium.

[0123] CC16-derived peptide mimics provide protection against human rhinovirus infection in airway epithelial cells. Preliminary studies were conducted in primary rhinovirus cells grown at the ALI. Transwell cells were stimulated with RV (multiple of infection (MOI) 0.1) for 4 hours, after which external RV was washed off the cell surface and the test compound was added; uninfected baseline controls were treated similarly by pipetting medium. Cells were allowed to incubate for another 48 hours. Three readings were used for analysis in this screening: RV infectivity, INF-λ induction, and TNF-α induction. The optimal compound for this screening would have reduced RV infection, maintain high levels of INF-λ, and have low levels of TNF-α.

[0124] The compound was tested in triplicate at an initial dose of 2.5 µM, and this initial dose was found to be optimal for inhibiting rCC16 infection by 50%. Figure 10 As shown, examples of RV screening demonstrate that the standard control [rCC16 from R&D and rCC16 prepared at UA (MJ)] and the active parent compound C836 exhibit activity in reducing RV infectivity. Those compounds with RV inhibitory activity are shown in green (top panel): 970, 971, 972, and 974. In the middle panel, among these compounds, compounds 972 and 974 maintain INF-λ levels. However, compound 974 exhibits a spike in TNF-α (bottom panel). In this screening against three factors of interest, compound 972 is the best candidate for advancing further studies. Compounds in gray are considered inactive in this assay. RV loading, INF-λ, and TNF-α were assessed by RT-PCR. These compounds advancing further studies will be tested in dose-response assays to determine IC50. 50 The required optimal concentration.

[0125] As used in this article, the term “about” refers to plus or minus 10% of the reference figure.

[0126] While preferred embodiments of the invention have been shown and described, it will be readily apparent to those skilled in the art that modifications can be made therein without departing from the scope of the appended claims. Therefore, the scope of the invention is limited only by the appended claims. In some embodiments, the drawings presented in this patent application are drawn to scale, including ratios of angles, dimensions, etc. In some embodiments, the drawings are merely representative, and the claims are not limited by the dimensions of the drawings. In some embodiments, the description of the invention, using the phrase "comprising," includes embodiments that can be described as "substantially consisting of" or "consisting of," and thus satisfies the written description requirements for claiming one or more embodiments of the invention using the phrase "substantially consisting of" or "consisting of."

Claims

1. A composition comprising a CC16-derived analogue.

2. The composition of claim 1, wherein the CC16-derived analogue comprises compound C836 according to the sequence DQD-Nle-REAGAQLKKLVDT (SEQ ID NO: 41).

3. The composition according to claim 1 or claim 2, wherein the CC16-derived analogue comprises the sequences according to SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40 and SEQ ID NO:

41.

4. The composition according to any one of claims 1 to 3, wherein the CC16-derived analogue comprises a sequence that is at least 95%, 90%, or 85% identical to SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, and SEQ ID NO:

41.

5. The composition according to any one of claims 1 to 4, wherein the CC16-derived analogue comprises N-terminal acetylation.

6. The composition according to any one of claims 1 to 5, wherein the CC16-derived analogue comprises C-terminal amidation.

7. The composition according to any one of claims 1 to 6, wherein the CC16-derived analogue is helical.

8. The composition according to any one of claims 1 to 7, wherein at least a portion of the CC16-derived analogue is cyclic.

9. The composition according to any one of claims 1 to 8, wherein the CC16-derived analogue retains activity similar to that of the full-length CC16 protein.

10. The composition according to any one of claims 1 to 9, wherein the composition is used to treat inflammatory, neoplastic and obstructive lung diseases.

11. The composition according to any one of claims 1 to 10, wherein the composition is in a formulation for nebulization, subcutaneous injection or pulmonary delivery.

12. The composition according to any one of claims 1 to 11, wherein the composition is administered via nasal inhalation, subcutaneous administration or oral administration.

13. A composition comprising a CC16-derived analogue according to the sequence DQD-Nle-REAGAQLKKLVDT (SEQ ID NO: 41).

14. The composition of claim 13, wherein the CC16-derived analogue comprises N-terminal acetylation.

15. The composition of claim 13 or claim 14, wherein the CC16-derived analog comprises C-terminal amidation.

16. The composition according to any one of claims 13 to 15, wherein the CC16-derived analogue is helical.

17. The composition according to any one of claims 13 to 16, wherein at least a portion of the CC16-derived analogue is cyclic.

18. The composition according to any one of claims 13 to 17, wherein the CC16-derived analogue retains activity similar to that of the full-length CC16 protein.

19. The composition according to any one of claims 13 to 18, wherein the composition is used to treat inflammatory, neoplastic and obstructive lung diseases.

20. The composition according to any one of claims 13 to 19, wherein the composition is in a formulation for nebulization, subcutaneous injection or pulmonary delivery.

21. The composition according to any one of claims 13 to 20, wherein the composition is administered via nasal inhalation, subcutaneous administration or oral administration.

22. A method of treating inflammatory, neoplastic, and obstructive lung diseases in a subject with such need, the method comprising administering an effective amount of a CC16-derived analogue to the patient.

23. The method of claim 22, wherein the CC16-derived analogue comprises compound C836 according to the sequence DQD-Nle-REAGAQLKKLVDT (SEQ ID NO: 41).

24. The method of claim 22 or claim 23, wherein the CC16-derived analogue comprises sequences according to SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40 and SEQ ID NO:

41.

25. The method according to any one of claims 22 to 24, wherein the CC16-derived analogue comprises a sequence that is at least 95%, 90%, or 85% identical to SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, and SEQ ID NO:

41.

26. The method of any one of claims 22 to 25, wherein the CC16-derived analogue comprises N-terminal acetylation.

27. The method according to any one of claims 22 to 26, wherein the CC16-derived analogue comprises C-terminal amidation.

28. The method according to any one of claims 22 to 27, wherein the CC16-derived analogue is helical.

29. The method according to any one of claims 22 to 28, wherein at least a portion of the CC16-derived analogue is cyclic.

30. The method according to any one of claims 22 to 28, wherein the CC16-derived analogue retains activity similar to the full-length CC16 protein.

31. The method according to any one of claims 22 to 30, wherein the composition is used to treat inflammatory, neoplastic and obstructive lung diseases.

32. The method according to any one of claims 22 to 31, wherein the composition is in a formulation for nebulization, subcutaneous injection or pulmonary delivery.

33. The method according to any one of claims 22 to 32, wherein the composition is administered via nasal inhalation, subcutaneous administration, or oral administration.

34. A method of treating inflammatory, neoplastic, and obstructive lung diseases in a subject with such need, the method comprising administering to the patient an effective amount of a CC16-derived analog according to the sequence DQD-Nle-REAGAQLKKLVDT (SEQ ID NO: 41).

35. The method of claim 34, wherein the CC16-derived analogue comprises N-terminal acetylation.

36. The method of claim 34 or claim 35, wherein the CC16-derived analog comprises C-terminal amidation.

37. The method according to any one of claims 34 to 36, wherein the CC16-derived analogue is helical.

38. The method according to any one of claims 34 to 36, wherein at least a portion of the CC16-derived analogue is cyclic.

39. The method according to any one of claims 34 to 38, wherein the CC16-derived analogue retains activity similar to the full-length CC16 protein.

40. The method according to any one of claims 34 to 39, wherein the composition is used to treat inflammatory, neoplastic and obstructive lung diseases.

41. The method according to any one of claims 34 to 40, wherein the composition is in a formulation for nebulization, subcutaneous injection or pulmonary delivery.

42. The method according to any one of claims 34 to 41, wherein the composition is administered via nasal inhalation, subcutaneous administration, or oral administration.

43. A CC16-derived analog composition for use in the treatment of inflammatory, neoplastic, and obstructive pulmonary diseases in subjects with such need.

44. The composition of claim 43, wherein the CC16-derived analog comprises compound C836 according to the sequence DQD-Nle-REAGAQLKKLVDT (SEQ ID NO: 41).

45. The composition according to claim 43 or claim 44, wherein the CC16-derived analogue comprises the sequence according to SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40 and SEQ ID NO:

41.

46. ​​A CC16-derived analog composition for use in treating inflammatory, neoplastic, and obstructive pulmonary diseases in subjects with such need; wherein the CC16-derived analog composition comprises the sequence according to SEQ ID NO:

41.

47. The composition according to any one of claims 43 to 46, wherein the CC16-derived analogue comprises at least 95%, 90%, or 85% identical sequences to SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, and SEQ ID NO:

41.

48. The composition according to any one of claims 43 to 47, wherein the CC16-derived analogue comprises N-terminal acetylation.

49. The composition according to any one of claims 43 to 48, wherein the CC16-derived analog comprises C-terminal amidation.

50. The composition according to any one of claims 43 to 49, wherein the CC16-derived analogue is helical.

51. The composition according to any one of claims 43 to 49, wherein at least a portion of the CC16-derived analogue is cyclic.

52. The composition according to any one of claims 43 to 51, wherein the CC16-derived analogue retains activity similar to that of the full-length CC16 protein.

53. The composition according to any one of claims 43 to 52, wherein the composition is used to treat inflammatory, neoplastic and obstructive lung diseases.

54. The composition according to any one of claims 43 to 53, wherein the composition is in a formulation for nebulization, subcutaneous injection or pulmonary delivery.

55. The composition according to any one of claims 43 to 54, wherein the composition is administered via nasal inhalation, subcutaneous administration, or oral administration.

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