New molecules

Abstract

The present invention relates to TLR2 agonist compounds and compositions thereof, as well as the use of such compounds and compositions in preventing and / or treating respiratory tract infections or diseases or conditions associated with viral or bacterial infections.

Description

[0001] This application claims priority to Australian Provisional Patent Application No. 2019902231 (filed on 26 June 2019) and Australian Provisional Patent Application No. 2019904862 (filed on 20 December 2019). The entire contents of each of AU 2019902231 and AU 2019904862 are incorporated herein by reference. Technical Field

[0002] The present invention relates to compounds and compositions thereof, and the use of such compounds and compositions in preventing and / or treating respiratory tract infections or respiratory diseases or conditions associated with viral or bacterial infections. Background Art

[0003] Respiratory infections are the most common cause of human illness worldwide and are often caused by viruses. According to the World Health Organization (WHO), seasonal influenza epidemics alone cause approximately 3 to 5 million severe illnesses and 250,000 to 500,000 deaths worldwide each year.

[0004] Although vaccines can be used for some seasonal strains, such as influenza, these vaccines have not always been shown to be sufficient due to several factors (such as infection during the lag period between vaccination and the formation of antibodies and immune cells). Seasonal vaccinations also typically require changes, including reformulation and administration, and thus may not provide protection over the entire time span of expectation. For other influenza events, such as unexpected full-scale outbreaks, vaccines are not always known, developed, or available.

[0005] Viral respiratory infections can also exacerbate the severity of respiratory conditions, leading to exacerbations (attacks). Conditions such as asthma and chronic obstructive pulmonary disease (COPD) can experience exacerbations. Asthma and COPD exacerbations are the most clinically and economically important forms of these diseases.

[0006] Despite the best available treatments, the vast majority of exacerbations (particularly for asthma) continue to occur. When an exacerbation does occur, treatment options are limited and have barely evolved in recent years. Treatment involves increasing the dose of inhaled bronchodilators and systemic or oral corticosteroids, the same medications that failed to prevent the exacerbation in the first place.

[0007] Therefore, there is a need for new or improved compounds and methods for treating and / or preventing respiratory tract infections or respiratory tract conditions associated with viral or bacterial infections.

[0008] The reference to any prior art in this specification is not an acknowledgment or suggestion that this prior art forms part of the common general knowledge in any jurisdiction, or that this prior art could reasonably be expected to be understood as being related to and / or combinable with other prior art known to the person skilled in the art. Summary of the Invention

[0009] The present invention provides Toll-like receptor 2 protein (TLR2) agonist compounds and compositions thereof. TLR2 agonists have previously been identified as showing potential for treating respiratory diseases and conditions associated with infectious agents (such as viruses and bacteria). Advantageously, the compounds and compositions of the present application can show activity and have been used in therapeutic areas, such as treating and / or preventing respiratory diseases or conditions associated with viral or bacterial infections. In addition, the compounds and compositions of the present application can exhibit increased stability, which can be converted into longer clearance rates after administration. Compared to other related compounds, the compounds of the present invention exhibit improved solution stability.

[0010] In one aspect, the present invention provides a compound comprising the following structure:

[0011] AYB

[0012] Where A contains or consists of the following structures:

[0013]

[0014] in

[0015] b and w are each independently an integer from 0 to 7, and v is an integer from 0 to 5, such as from 2 to 5, with the proviso that:

[0016] The sum of b, v, and w is at least 3; and

[0017] The sum of b and w ranges from 0 to 7;

[0018] z is 1 or 2;

[0019] X is selected from -S-, -S(=O)- and -S(=O)2-;

[0020] Z1 and Z2 are each independently selected from the group consisting of: -O-, -NR-, -S-, S(=O),

[0021] -S(=O)2-, -C(=O)O-, -OC(=O)-, -C(=O)NR-, -NRC(=O)-, -C(=O)S-, -SC(=O)-, -OC(=O)O-, -NRC(=O)O-, -OC(=O)NR- and -NRC(=O)NR-;

[0022] R 11 、R 12 、R x 、R y 、R 14 、R 15 、R 16 and R 17 Each independently represents H or a C1-C6 aliphatic group;

[0023] R, R 13 and R 18 Each independently is H or a C1-C6 aliphatic group;

[0024] R 19 is H, a C1-C6 aliphatic group, an amino protecting group, L3-C(=O)-, or A2;

[0025] L1 and L2 are each independently C5-C 21 Aliphatic or C4-C 20 heteroaliphatic groups;

[0026] L3 is C1-C 21 Aliphatic or C2-C 20 heteroaliphatic groups;

[0027] A2 is an amino acid or peptide;

[0028] Among them, there are 11 、R 12 、R 13 、R 14 、R 15 、R 16 、R 17 、R 18 、R 19 、R x 、R y Any aliphatic or heteroaliphatic group in any of L1, L2, and L3 is optionally substituted;

[0029] Y is

[0030]

[0031] wherein R1 and R2 are independently selected from the group consisting of H, -CH2OH, -CH2CH2OH, -CH(CH3)OH, -CH2OPO(OH)2, -CH2C(=O)NH2, -CH2CH2C(=O)OH, and -CH2CH2C(=O)OR8, wherein any of the alkyl hydrogens may be substituted by halogen;

[0032] R8 is selected from the group consisting of H and linear or branched C1-C6 alkyl;

[0033] and

[0034] B comprises polyethylene glycol (PEG) or consists of polyethylene glycol (PEG),

[0035] or a pharmaceutically acceptable salt, solvate or prodrug thereof.

[0036] The present invention also provides a compound comprising A and PEG, wherein A and PEG are linked via a glycine, serine, homoserine, threonine, phosphoserine, asparagine or glutamine residue, or an ester of a glutamine residue,

[0037] in

[0038] A in this compound has the following structure:

[0039]

[0040] in

[0041] b and w are each independently an integer from 0 to 7, and v is an integer from 0 to 5, such as from 2 to 5, with the proviso that:

[0042] The sum of b, v, and w is at least 3; and

[0043] The sum of b and w ranges from 0 to 7;

[0044] z is 1 or 2;

[0045] X is selected from -S-, -S(=O)- and -S(=O)2-;

[0046] Z1 and Z2 are each independently selected from the group consisting of: -O-, -NR-, -S-, S(=O), -S(=O)2-, -C(=O)O-, -OC(=O)-, -C(=O)NR-, -NRC(=O)-, -C(=O)S-, -SC(=O)-, -OC(=O)O-, -NRC(=O)O-, -OC(=O)NR- and -NRC(=O)NR-;

[0047] In each case of b, v, w, and z, R 11 、R 12 、R x 、R y 、R 14 、R 15 、R 16 and R 17 Each independently is H or a C1-C6 aliphatic group;

[0048] R, R 13 and R 18Each independently is H or a C1-C6 aliphatic group;

[0049] R 19 is H, a C1-C6 aliphatic group, an amino protecting group, L3-C(=O)-, or A2;

[0050] L1 and L2 are each independently C5-C 21 Aliphatic or C4-C 20 heteroaliphatic groups;

[0051] L3 is C1-C 21 Aliphatic or C2-C 20 heteroaliphatic groups;

[0052] A2 is an amino acid or peptide;

[0053] Among them, there are 11 、R 12 、R 13 、R 14 、R 15 、R 16 、R 17 、R 18 、R 19 、R x 、R y Any aliphatic or heteroaliphatic group in any of L1, L2, and L3 is optionally substituted;

[0054] or a pharmaceutically acceptable salt, solvate or prodrug thereof.

[0055] In one aspect, the present invention provides a compound comprising the following structure:

[0056]

[0057] wherein R1 and R2 are independently selected from the group consisting of H, -CH2OH, -CH2CH2OH, -CH(CH3)OH, -CH2OPO(OH)2, -CH2C(=O)NH2, -CH2CH2C(=O)OH, and -CH2CH2C(=O)OR8, wherein any of the alkyl hydrogens may be substituted by halogen;

[0058] R8 is selected from the group consisting of H and linear or branched C1-C6 alkyl;

[0059] b and w are each independently an integer from 0 to 7, and v is an integer from 0 to 5, provided that:

[0060] The sum of b, v, and w is at least 3; and

[0061] The sum of b and w ranges from 0 to 7;

[0062] z is 1 or 2;

[0063] X is selected from -S-, -S(=O)- and -S(=O)2-;

[0064] Z1 and Z2 are each independently selected from the group consisting of: -O-, -NR-, -S-, -S(=O)-, -S(=O)2-, -C(=O)O-, -OC(=O)-, -C(=O)NR-, -NRC(=O)-, -C(=O)S-, -SC(=O)-, -OC(=O)O-, -NRC(=O)O-, -OC(=O)NR-, and -NRC(=O)NR-;

[0065] In each case of b, v, w, and z, R 11 、R 12 、R x 、R y 、R 14 、R 15 、R 16 and R 17 Each independently is H or a C1-C6 aliphatic group;

[0066] R, R 13 and R 18 Each independently is H or a C1-C6 aliphatic group;

[0067] R 19 is H, a C1-C6 aliphatic group, an amino protecting group, L3-C(=O)-, or A2;

[0068] L1 and L2 are each independently C5-C 21 Aliphatic or C4-C 20 heteroaliphatic groups;

[0069] L3 is C1-C 21 Aliphatic or C2-C 20 heteroaliphatic groups;

[0070] A2 is an amino acid or peptide;

[0071] Among them, there are 11 、R 12 、R 13 、R 14 、R 15 、R 16 、R 17 、R 18 、R 19 、R x 、R yAny aliphatic or heteroaliphatic group in any of L1, L2, and L3 is optionally substituted;

[0072] The above structure is covalently linked to polyethylene glycol (PEG),

[0073] or a pharmaceutically acceptable salt, solvate or prodrug thereof.

[0074] In one aspect, the present invention provides a compound having formula (VI):

[0075]

[0076] in

[0077] n is 3 to 100;

[0078] m is 1, 2, 3 or 4;

[0079] p is 2, 3, or 4;

[0080] q is zero or 1;

[0081] R1 and R2 are independently selected from the group consisting of H, -CH2OH, -CH2CH2OH, -CH(CH3)OH, -CH2OPO(OH)2, -CH2C(=O)NH2, -CH2CH2C(=O)OH, and -CH2CH2C(=O)OR8, wherein any of the alkyl hydrogens may be substituted by halogen;

[0082] R8 is selected from the group consisting of H and linear or branched C1-C6 alkyl;

[0083] wherein when q=1, R3 is -NH2 or -OH;

[0084] Wherein when q=0, R3 is H;

[0085] L is zero or consists of 1 to 10 units, each of which is a natural alpha amino acid or derived from a natural alpha amino acid and has the formula:

[0086]

[0087] wherein R4 is H; and

[0088] R5 is the side chain, or the second hydrogen of the amino acid,

[0089] b and w are each independently an integer from 0 to 7, and v is an integer from 0 to 5, provided that:

[0090] The sum of b, v, and w is at least 3; and

[0091] The sum of b and w ranges from 0 to 7;

[0092] z is 1 or 2;

[0093] X is selected from -S-, -S(=O)- and -S(=O)2-;

[0094] Z1 and Z2 are each independently selected from the group consisting of: -O-, -NR-, -S-, -S(=O)-, -S(=O)2-, -C(=O)O-, -OC(=O)-, -C(=O)NR-, -NRC(=O)-, -C(=O)S-, -SC(=O)-, -OC(=O)O-, -NRC(=O)O-, -OC(=O)NR-, and -NRC(=O)NR-;

[0095] In each case of b, v, w, and z, R 11 、R 12 、R x 、R y 、R 14 、R 15 、R 16 and R 17 Each independently is H or a C1-C6 aliphatic group;

[0096] R, R 13 and R 18 Each independently is H or a C1-C6 aliphatic group;

[0097] R 19 is H, a C1-C6 aliphatic group, an amino protecting group, L3-C(=O)-, or A2;

[0098] L1 and L2 are each independently C5-C 21 Aliphatic or C4-C 20 heteroaliphatic groups;

[0099] L3 is C1-C 21 Aliphatic or C2-C 20 heteroaliphatic groups;

[0100] A2 is an amino acid or peptide;

[0101] Among them, there are 11 、R 12 、R 13 、R 14 、R 15 、R 16 、R 17 、R 18 、R 19 、R x 、R yAny aliphatic or heteroaliphatic group in any of L1, L2, and L3 is optionally substituted;

[0102] or a pharmaceutically acceptable salt, solvate or prodrug thereof.

[0103] In one embodiment, the present invention provides a compound having formula (VII):

[0104] AY-NH-(CH2) p -O-(CH2-CH2-O) n -[(CH2) m -CO-L-] q R3

[0105] (VII)

[0106] in

[0107] A has the following structure:

[0108]

[0109] Y is

[0110]

[0111] wherein R1 and R2 are independently selected from the group consisting of H, -CH2OH, -CH2CH2OH, -CH(CH3)OH, -CH2OPO(OH)2, -CH2C(=O)NH2, -CH2CH2C(=O)OH, and -CH2CH2C(=O)OR8, wherein any of the alkyl hydrogens may be substituted by halogen;

[0112] R8 is selected from the group consisting of H and linear or branched C1-C6 alkyl;

[0113] n is 3 to 100;

[0114] m is 1, 2, 3 or 4;

[0115] p is 2, 3, or 4;

[0116] q is zero or 1;

[0117] wherein when q=1, R3 is -NH2 or -OH;

[0118] Wherein when q=0, R3 is H;

[0119] L is zero or consists of 1 to 10 units, each of which is a natural alpha amino acid or derived from a natural alpha amino acid and has the formula:

[0120]

[0121] wherein R4 is H; and

[0122] R5 is the side chain, or the second hydrogen of the amino acid,

[0123] b and w are each independently an integer from 0 to 7, and v is an integer from 0 to 5, provided that:

[0124] The sum of b, v, and w is at least 3; and

[0125] The sum of b and w ranges from 0 to 7;

[0126] z is 1 or 2;

[0127] X is selected from -S-, -S(=O)- and -S(=O)2-;

[0128] Z1 and Z2 are each independently selected from the group consisting of: -O-, -NR-, -S-, -S(=O)-, -S(=O)2-, -C(=O)O-, -OC(=O)-, -C(=O)NR-, -NRC(=O)-, -C(=O)S-, -SC(=O)-, -OC(=O)O-, -NRC(=O)O-, -OC(=O)NR-, and -NRC(=O)NR-;

[0129] In each case of b, v, w, and z, R 11 、R 12 、R x 、R y 、R 14 、R 15 、R 16 and R 17 Each independently is H or a C1-C6 aliphatic group;

[0130] R, R 13 and R 18 Each independently is H or a C1-C6 aliphatic group;

[0131] R 19 is H, a C1-C6 aliphatic group, an amino protecting group, L3-C(=O)-, or A2;

[0132] L1 and L2 are each independently C5-C 21 Aliphatic or C4-C 20 heteroaliphatic groups;

[0133] L3 is C1-C 21 Aliphatic or C2-C 20 heteroaliphatic groups;

[0134] A2 is an amino acid or peptide;

[0135] Among them, there are 11 、R 12 、R 13 、R 14 、R 15 、R 16 、R 17 、R 18 、R 19 、R x 、R y Any aliphatic or heteroaliphatic group in any of L1, L2, and L3 is optionally substituted;

[0136] or a pharmaceutically acceptable salt, solvate or prodrug thereof. In one embodiment, the compound has formula (X):

[0137]

[0138] in

[0139] n is 3 to 100;

[0140] k is 3 to 100;

[0141] m is 1, 2, 3 or 4;

[0142] p is 2, 3, or 4;

[0143] t is 2, 3, or 4;

[0144] h is 1, 2, 3, or 4;

[0145] q is zero or 1;

[0146] wherein R1 and R2 are independently selected from the group consisting of H, -CH2OH, -CH2CH2OH, -CH(CH3)OH, -CH2OPO(OH)2, -CH2C(=O)NH2, -CH2CH2C(=O)OH, and -CH2CH2C(=O)OR8, wherein any of the alkyl hydrogens may be substituted by halogen;

[0147] R8 is selected from the group consisting of H and linear or branched C1-C6 alkyl;

[0148] wherein when q=1, R3 is -NH2 or -OH;

[0149] Wherein when q=0, R3 is H;

[0150] L is zero or consists of 1 to 10 units, each of which is a natural alpha amino acid or derived from a natural alpha amino acid and has the formula:

[0151]

[0152] wherein R4 is H; and

[0153] R5 is the side chain, or the second hydrogen of the amino acid,

[0154] b and w are each independently an integer from 0 to 7, and v is an integer from 0 to 5, provided that:

[0155] The sum of b, v, and w is at least 3; and

[0156] The sum of b and w ranges from 0 to 7;

[0157] z is 1 or 2;

[0158] X is selected from -S-, -S(=O)- and -S(=O)2-;

[0159] Z1 and Z2 are each independently selected from the group consisting of: -O-, -NR-, -S-, -S(=O)-, -S(=O)2-, -C(=O)O-, -OC(=O)-, -C(=O)NR-, -NRC(=O)-, -C(=O)S-, -SC(=O)-, -OC(=O)O-, -NRC(=O)O-, -OC(=O)NR-, and -NRC(=O)NR-;

[0160] In each case of b, v, w, and z, R 11 、R 12 、R x 、R y 、R 14 、R 15 、R 16 and R 17 Each independently is H or a C1-C6 aliphatic group;

[0161] R, R 13 and R 18 Each independently is H or a C1-C6 aliphatic group;

[0162] R 19 is H, a C1-C6 aliphatic group, an amino protecting group, L3-C(=O)-, or A2;

[0163] L1 and L2 are each independently C5-C 21 Aliphatic or C4-C 20 heteroaliphatic groups;

[0164] L3 is C1-C 21 Aliphatic or C2-C 20 heteroaliphatic groups;

[0165] A2 is an amino acid or peptide;

[0166] Among them, there are 11 、R 12 、R 13 、R 14 、R 15 、R 16 、R 17 、R 18 、R 19 、R x 、R y Any aliphatic or heteroaliphatic group in any of L1, L2, and L3 is optionally substituted;

[0167] or a pharmaceutically acceptable salt, solvate or prodrug thereof.

[0168] The present invention also provides compositions comprising, consisting essentially of, or consisting of a compound of the present invention as described herein, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, and a pharmaceutically acceptable carrier, diluent, or excipient.

[0169] In one aspect, the present invention provides a method of treating and / or preventing a disease, comprising enhancing the innate immune response in a subject by administering to a subject in need thereof an effective amount of a compound of the present invention as described herein, or a pharmaceutically acceptable salt, solvate or prodrug thereof.

[0170] In another aspect, the present invention provides a method for treating and / or preventing a disease associated with or caused by an infectious agent, comprising administering to a subject in need thereof an effective amount of a compound of the present invention as described herein, or a pharmaceutically acceptable salt, solvate or prodrug thereof.

[0171] In another aspect, the present invention provides a method for treating and / or preventing respiratory diseases or conditions associated with viral or bacterial infections, comprising administering to a subject in need thereof a compound of the present invention as described herein, or a pharmaceutically acceptable salt, solvate or prodrug thereof.

[0172] In another aspect, the present invention provides a method for treating and / or preventing respiratory tract infections, comprising administering to a subject in need thereof a compound of the present invention as described herein, or a pharmaceutically acceptable salt, solvate or prodrug thereof.

[0173] In another aspect, the present invention provides a method for reducing airway inflammation, comprising administering to a subject in need thereof a compound of the present invention as described herein, or a pharmaceutically acceptable salt, solvate, or prodrug thereof.

[0174] The present invention also provides a method for improving a subject's ability to control a respiratory disease or condition during a respiratory viral infection, the method comprising administering to a subject in need thereof a compound of the present invention as described herein, or a pharmaceutically acceptable salt, solvate or prodrug thereof.

[0175] The present invention also provides a method for treating and / or preventing diseases or conditions associated with TLR2 receptor, comprising administering to a subject in need thereof a compound of the present invention as described herein or a pharmaceutically acceptable salt, solvate or prodrug thereof.

[0176] The present invention also provides a method of agonizing TLR2 activity in a cell, comprising contacting the cell with a compound of the present invention as described herein, or a pharmaceutically acceptable salt, solvate, or prodrug thereof.

[0177] In another aspect, the present invention provides use of a compound of the present invention as described herein, or a pharmaceutically acceptable salt, solvate or prodrug thereof, in the preparation of a medicament for enhancing an innate immune response in a subject.

[0178] In another aspect, the present invention provides the use of a compound of the present invention as described herein, or a pharmaceutically acceptable salt, solvate or prodrug thereof, in the preparation of a medicament for treating and / or preventing a disease caused by an infectious agent.

[0179] In another aspect, the present invention further provides the use of a compound of the present invention as described herein, or a pharmaceutically acceptable salt, solvate or prodrug thereof, in the preparation of a medicament for treating and / or preventing a respiratory disease or condition associated with a viral or bacterial infection in a subject.

[0180] In another aspect, the present invention further provides use of a compound of the present invention as described herein, or a pharmaceutically acceptable salt, solvate or prodrug thereof, in the preparation of a medicament for treating and / or preventing a respiratory tract infection in a subject.

[0181] In another aspect, the present invention further provides use of a compound of the present invention as described herein, or a pharmaceutically acceptable salt, solvate or prodrug thereof, in the preparation of a medicament for reducing airway inflammation.

[0182] In another aspect, the present invention further provides the use of a compound of the present invention as described herein, or a pharmaceutically acceptable salt, solvate or prodrug thereof, in the preparation of a medicament for improving a subject's ability to control a respiratory disease or condition during a respiratory viral infection.

[0183] In another aspect, the present invention further provides use of a compound of the present invention as described herein, or a pharmaceutically acceptable salt, solvate or prodrug thereof, in the preparation of a medicament for treating and / or preventing a disease or condition associated with the TLR2 receptor.

[0184] In one aspect, the present invention provides the use of a compound of the present invention as described herein, or a pharmaceutically acceptable salt, solvate or prodrug thereof, for enhancing an innate immune response in a subject.

[0185] In another aspect, the present invention provides the use of a compound of the present invention as described herein, or a pharmaceutically acceptable salt, solvate or prodrug thereof, for preventing a disease caused by an infectious agent in a subject.

[0186] In another aspect, the present invention provides the use of a compound of the present invention as described herein, or a pharmaceutically acceptable salt, solvate or prodrug thereof, for treating and / or preventing a respiratory disease or disorder associated with a viral or bacterial infection in a subject.

[0187] In another aspect, the present invention provides the use of a compound of the present invention as described herein, or a pharmaceutically acceptable salt, solvate or prodrug thereof, for reducing airway inflammation in a subject.

[0188] In another aspect, the present invention provides the use of a compound of the present invention as described herein, or a pharmaceutically acceptable salt, solvate or prodrug thereof, for managing a respiratory disease or disorder during a respiratory viral infection in a subject.

[0189] In another aspect, the present invention provides the use of a compound of the present invention as described herein, or a pharmaceutically acceptable salt, solvate or prodrug thereof, for treating and / or preventing a disease or condition associated with the TLR2 receptor.

[0190] In another aspect, the present invention further provides use of a compound of the present invention as described herein, or a pharmaceutically acceptable salt, solvate or prodrug thereof, for agonizing TLR2 in a cell.

[0191] The present invention also provides a kit for use in the method of the present invention, or a kit when used in the method of the present invention, the kit comprising, consisting essentially of, or consisting of:

[0192] - a compound of the invention as described herein; and optionally

[0193] - written instructions describing the use of the compound in the methods of the invention.

[0194] In yet another aspect, the present invention provides a method for preparing a compound of the present invention. In some embodiments, the method comprises the steps outlined in the synthesis shown in the examples.

[0195] Further aspects of the invention described in the preceding paragraphs and further embodiments of these aspects will become apparent from the following description, given by way of example and with reference to the accompanying drawings. BRIEF DESCRIPTION OF THE DRAWINGS

[0196] Figure 1 TLR2 activity of compounds 3, 4, 15 and 16 from the NK-κB luciferase assay described in Example 2.

[0197] Figure 2 .Stability results of compounds 3, 4, 15 and 16 after storage in saline (0.9%) or phosphate buffered saline (PBS, pH 7.4) at 25°C or 40°C for two weeks (14 days) relative to the percentage peak area of ​​the main HPLC peak at time zero, which was normalized to 100% to exclude the contribution of impurities to the total peak area.

[0198] Figure 3 .Generated using the main peak area Figure 2 The two-week (14-day) stability results of the main peak are shown in Figure 2, and the main peak area is expressed as a percentage of the area at time zero.

[0199] Figure 4 TLR2 activity of compounds 4, 20, 24 and 36 from the NK-κB luciferase assay described in Example 9. DETAILED DESCRIPTION

[0200] It should be understood that the invention disclosed and defined in this specification extends to all alternative combinations of two or more of the individual features mentioned or evident from the text and drawings. All these different combinations constitute multiple alternative aspects of the invention.

[0201] Reference will now be made in detail to certain embodiments of the present invention. Although the present invention will be described in conjunction with the embodiments, it should be understood that the present invention is not intended to be limited to those embodiments. On the contrary, the present invention is intended to encompass all alternatives, modifications, and equivalents that may be included within the scope of the present invention as defined in the claims.

[0202] Those skilled in the art will recognize that many methods and materials similar to or equivalent to those described herein can be used in the practice of the present invention. The present invention is in no way limited to the methods and materials described. It should be understood that the invention disclosed and defined in this specification extends to all alternative combinations of two or more of the individual features mentioned or apparent from the text and drawings. All of these different combinations constitute multiple alternative aspects of the present invention.

[0203] All patents and publications mentioned herein are incorporated by reference in their entirety.

[0204] As used herein, the term "comprise" and variations of the term such as "includes" and "comprising" are not intended to exclude further additives, components, integers or steps, unless the context requires otherwise.

[0205] For the purpose of interpreting this specification, terms used in the singular also include the plural and vice versa.

[0206] General chemical terms used in the formulae herein have their ordinary meanings.

[0207] The term "aliphatic" is intended to include saturated and unsaturated, non-aromatic, straight-chain, branched, acyclic and cyclic hydrocarbons. It will be appreciated by those skilled in the art that aliphatic groups include, for example, alkyl, alkenyl, alkynyl, cycloalkyl and cycloalkenyl and their hybrids, such as (cycloalkyl) alkyl, (cycloalkenyl) alkyl and (cycloalkyl) alkenyl. In different embodiments, aliphatic groups include from 1 to 12, 1 to 8, 1 to 6 or 1 to 4 carbon atoms. In certain embodiments, aliphatic groups include 5 to 21, from 9 to 21 or from 11 to 21 carbon atoms, such as from 11, 13, 15, 17 or 19 carbon atoms. In certain embodiments, aliphatic groups are saturated.

[0208] The term "heteroaliphatic" is intended to include aliphatic groups in which one or more chain and / or ring carbon atoms are independently substituted with heteroatoms, preferably heteroatoms selected from oxygen, nitrogen, and sulfur. In some embodiments, heteroaliphatic groups are saturated. Examples of heteroaliphatic groups include straight or branched heteroalkyl, heteroalkenyl, and heteroalkynyl groups.

[0209] The term "alkyl" is intended to include saturated straight and branched chain hydrocarbon groups. In some embodiments, the alkyl group has from 1 to 12, 1 to 10, 1 to 8, 1 to 6, or from 1 to 4 carbon atoms. In some embodiments, the alkyl group has from 5 to 21, from 9 to 21, or from 11 to 21 carbon atoms, such as from 11, 13, 15, 17, or 19 carbon atoms. Examples of straight chain alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl, and n-octyl. Examples of branched chain alkyl groups include, but are not limited to, isopropyl, isobutyl, sec-butyl, tert-butyl, neopentyl, isopentyl, and 2,2-dimethylpropyl.

[0210] The term "alkenyl" is intended to include straight and branched alkyl groups having at least one double bond between two carbon atoms. In some embodiments, the alkenyl group has from 2 to 12, from 2 to 10, from 2 to 8, from 2 to 6, or from 2 to 4 carbon atoms. In some embodiments, the alkenyl group has from 5 to 21, from 9 to 21, or from 11 to 21 carbon atoms, such as from 11, 13, 15, 17, or 19 carbon atoms. In some embodiments, the alkenyl group has one, two, or three carbon-carbon double bonds. Examples of alkenyl groups include, but are not limited to, vinyl, allyl, -CH=CH(CH ), -CH=C(CH ) , -C(CH )=CH , and -C(CH )=CH(CH ).

[0211] The term "alkynyl" is intended to include straight and branched alkyl groups having at least one triple bond between two carbon atoms. In some embodiments, the alkynyl group has from 2 to 12, from 2 to 10, from 2 to 8, from 2 to 6, or from 2 to 4 carbon atoms. In some embodiments, the alkynyl group has one, two, or three carbon-carbon triple bonds. Examples include, but are not limited to, -C=CH, -C=CH3, -CH2C=CH3, and -C=CH2CH(CH2CH3)2.

[0212] The term "heteroalkyl" is intended to include alkyl groups in which one or more of the chain carbon atoms are replaced by heteroatoms, preferably heteroatoms selected from the group consisting of oxygen, nitrogen, and sulfur. In certain embodiments, the heteroalkyl group is saturated. The heteroalkyl group includes, for example, polyethylene glycol groups and polyethylene glycol ether groups.

[0213] The term "cycloalkyl" is intended to include monocyclic, bicyclic, or tricyclic alkyl groups. In certain embodiments, one or more rings of the cycloalkyl group have from 3 to 12, from 3 to 10, from 3 to 8, from 3 to 6, or from 3 to 5 carbon atoms. In certain embodiments, the cycloalkyl group has 5 or 6 ring carbon atoms. Examples of monocyclic cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. In certain embodiments, the cycloalkyl group has from 3 to 8, from 3 to 7, from 3 to 6, from 4 to 6, from 3 to 5, or from 4 to 5 ring carbon atoms. Bicyclic and tricyclic ring systems include bridged, spirocyclic, and fused cycloalkyl ring systems. Examples of bicyclic and tricyclic cycloalkyl systems include, but are not limited to, bicyclo[2.1.1]hexyl, bicyclo[2.2.1]heptanyl, adamantyl, and decahydronaphthyl.

[0214] The term "cycloalkenyl" is intended to include non-aromatic cycloalkyl groups having at least one double bond between two carbon atoms. In certain embodiments, the cycloalkenyl has one, two, or three double bonds. In certain embodiments, the cycloalkenyl has from 4 to 14, from 5 to 14, from 5 to 10, from 5 to 8, or from 5 to 6 carbon atoms in one or more rings. In certain embodiments, the cycloalkenyl has 5,6,7, or 8 ring carbon atoms. Examples of cycloalkenyl include cyclohexenyl, cyclopentenyl, cyclohexadienyl, butadienyl, pentadienyl, and hexadienyl.

[0215] The term "aryl" is intended to include cyclic aromatic hydrocarbon groups that do not contain any ring heteroatoms. Aryl groups include monocyclic, bicyclic, and tricyclic ring systems. Examples of aryl groups include, but are not limited to, phenyl, azulenyl, heptalenyl, biphenyl, fluorenyl, phenanthrenyl, anthracenyl, indenyl, indanyl, pentalenyl, and naphthyl. In some embodiments, one or more rings of the aryl group have from 6 to 14, from 6 to 12, or from 6 to 10 carbon atoms. In some embodiments, the aryl group is phenyl or naphthyl. Aryl groups include aromatic-aliphatic fused ring systems. Examples include, but are not limited to, indanyl and tetrahydronaphthyl.

[0216] The term "heterocyclic radical" is intended to include non-aromatic ring systems containing 3 or more ring atoms, wherein one or more of these ring atoms are heteroatoms. In certain embodiments, the heteroatoms are nitrogen, oxygen, or sulfur. In certain embodiments, the heterocyclic radical contains one, two, three, or four heteroatoms. In certain embodiments, the heterocyclic radical includes monocyclic, bicyclic, and tricyclic rings having from 3 to 16, from 3 to 14, from 3 to 12, from 3 to 10, from 3 to 8, or from 3 to 6 ring atoms. The heterocyclic radical includes partially unsaturated and saturated ring systems, such as imidazolinyl and imidazolidinyl. The heterocyclic radical includes fused and bridged ring systems containing heteroatoms, such as quinuclidine. Heterocyclyl groups include, but are not limited to, aziridinyl, azetidinyl, azepanyl, diazepanyl, 1,3-dioxanyl, 1,3-dioxolanyl, isoxazolidinyl, morpholinyl, piperazinyl, piperidinyl, pyranyl, pyrazolidinyl, pyrrolinyl, pyrrolidinyl, tetrahydrofuranyl, tetrahydrothiophenyl, thiadiazolidinyl, and trithianyl.

[0217] The term "heteroaryl" is intended to include aromatic ring systems containing 5 or more ring atoms, wherein one or more of these ring atoms is a heteroatom. In some embodiments, the heteroatom is nitrogen, oxygen, or sulfur. In some embodiments, heteroaryl includes monocyclic, bicyclic, and tricyclic ring systems having from 5 to 16, from 5 to 14, from 5 to 12, from 5 to 10, from 5 to 8, or from 5 to 6 ring atoms. Heteroaryl groups include, but are not limited to, pyrrolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, thiazolyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, thiophenyl, benzothiophenyl, furanyl, benzofuranyl, indolyl, azetindolyl (pyrrolopyridinyl), indazolyl, benzimidazolyl, pyrazolopyridinyl, triazolopyridinyl, benzotriazolyl, benzoxazolyl, benzothiazolyl, imidazopyridinyl, isoxazolopyridinylxanthinyl, guaninyl, quinolyl, isoquinolyl, tetrahydroquinolyl, quinoxalinyl, and quinazolinyl. Heteroaryl groups include fused ring systems in which all rings are aromatic, such as indolyl, and fused ring systems in which only one ring is aromatic, such as 2,3-dihydroindolinyl.

[0218] The terms "halo" or "halogen" are intended to include F, Cl, Br, and I.

[0219] The term "heteroatom" is intended to include oxygen, nitrogen, sulfur, or phosphorus. In some embodiments, the heteroatom is selected from the group consisting of oxygen, nitrogen, and sulfur.

[0220] As used herein, the term "substituted" is intended to mean that one or more hydrogen atoms in the indicated group are replaced with one or more independently selected suitable substituents, provided that the normal valence of each atom to which the substituent or substituents are attached is not exceeded and that the substitution results in a stable compound. In some embodiments, the optional substituents in the compounds described herein include, but are not limited to, halogen, CN, NO2, OH, NH2, NHR 100 NR 100 R 200 、C 1-6 Halogenated alkyl, C 1-6 Haloalkoxy, C(O)NH2, C(O)NHR 100 、C(O)NR 100 R 200 、SO2R 100 , OR 100 SR 100 、S(O)R 100 、C(O)R 100 , and C 1-6 Aliphatic group; wherein R 100 and R 200 Each is independently C 1-6 Aliphatic groups, such as C 1-6 alkyl.

[0221] As used herein, the term "carboxyl protecting group" is intended to mean a group that can be readily removed to provide an OH group in the carboxyl group and that protects the carboxyl group from undesirable reactions during the synthesis procedure. Such protecting groups are described in Protective Groups in Organic Synthesis by TW Greene et al. (John Wiley & Sons, 1999) and in 'Amino Acid-Protecting Groups' by Fernando Albericio (with Albert Isidro-Llobet and Mercedes Alvarez), Chemical Reviews, 2009 (109) 2455-2504. Examples include, but are not limited to, alkyl and silyl groups, such as methyl, ethyl, tert-butyl, methoxymethyl, 2,2,2-trichloroethyl, benzyl, diphenylmethyl, trimethylsilyl, and tert-butyldimethylsilyl.

[0222] As used herein, the term "amine protecting group" is intended to mean a group that can be readily removed to provide an NH2 group in an amine group and that protects the amine group from undesirable reactions during synthetic procedures. Such protecting groups are described in Protective Groups in Organic Synthesis, edited by TW Greene et al. (John Wiley & Sons, 1999) and in 'Amino Acid-Protecting Groups' by Fernando Albericio (with Albert Isidro-Llobet and Mercedes Alvarez), Chemical Reviews, 2009 (109) 2455-2504. Examples include, but are not limited to, acyl and acyloxy groups such as acetyl, chloroacetyl, trichloroacetyl chloride, o-nitrophenylacetyl, o-nitrophenoxy-acetyl, trifluoroacetyl, acetoacetyl, 4-chlorobutyryl chloride, isobutyryl, picolinoyl, aminocaproyl, benzoyl, methoxy-carbonyl, 9-fluorenylmethoxycarbonyl, 2,2,2-trifluoroethoxycarbonyl, 2-trimethylsilylethoxy-carbonyl, tert-butyloxycarbonyl, benzyloxycarbonyl, p-nitrobenzyloxycarbonyl, 2,4-dichloro-benzyloxycarbonyl, etc. Additional examples include Cbz (carboxybenzyl), Nosyl ((o-nitrophenylsulfonyl or p-nitrophenylsulfonyl), Bpoc (2-(4-biphenylyl)isopropoxycarbonyl), and Dde (1-(4,4-dimethyl-2,6-dioxyhexylidene)ethyl).

[0223] As used herein, the term "carboxamide protecting group" is intended to mean a group that can be readily removed to provide the NH2 group in the carboxamide group and that protects the carboxamide group from undesirable reactions during synthetic procedures. Such protecting groups are described in Protective Groups in Organic Synthesis by TW Greene et al. (John Wiley & Sons, 1999) and in 'Amino Acid-Protecting Groups' by Fernando Albericio (with Albert Isidro-Llobet and Mercedes Alvarez), Chemical Reviews, 2009 (109) 2455-2504. Examples include, but are not limited to, 9-xanthenyl (Xan), trityl (Trt), methyltriphenyl (Mtt), dimethylcyclopropylmethyl (Cpd), and dimethylcyclopropylmethyl (Dmcp).

[0224] As used herein, the term "and / or" means "and" or "or", or both.

[0225] The term "one or more" preceding a noun contemplates both the singular and the plural, or both.

[0226] The term "ester" refers to a carboxylic acid group in which the hydrogen of the hydroxyl group has been replaced by a saturated straight-chain (i.e., linear) or branched hydrocarbon group. Specific examples of alkyl groups are methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, n-hexyl, and 2,2-dimethylbutyl. Alkyl groups may be C1-C6 alkyl groups. As used herein, the wording of the limits of a length range, such as "from 1 to 5," means any integer from 1 to 5, i.e., 1, 2, 3, 4, and 5. In other words, any range defined by two integers explicitly mentioned is intended to include and disclose any integer defining the limits and any integer contained in the range. Alkyl groups may be branched alkyl groups.

[0227] Reference to a numerical range disclosed herein (e.g., 1 to 10) is intended to also include reference to all rational numbers within that range (e.g., 1, 1.1, 2, 3, 3.9, 4, 5, 6, 6.5, 7, 8, 9, and 10), as well as any range of rational numbers within that range (e.g., 2 to 8, 1.5 to 5.5, and 3.1 to 4.7), and thus, all subranges of all ranges explicitly disclosed herein are hereby expressly disclosed. These are merely examples of what is specifically intended, and all possible combinations of numerical values ​​between the lowest and highest values ​​recited are to be considered to be expressly stated in this application in a similar manner.

[0228] As discussed above, the inventors have developed and optimized compounds for treating and / or preventing respiratory diseases or conditions, particularly those diseases or conditions associated with infectious agents such as bacteria or viruses. Specifically, when those compounds are applied to the upper respiratory tract, the compounds can provide significant protection against viral replication in the lungs. These compounds can have a higher efficacy than other known TLR2 agonists. In the animal models described herein, TLR2 agonist efficacy can also occur without significantly damaging TLR specificity and / or without causing significant weight loss. Further, these compounds can show favorable stability, for example, in formulations and / or biomatrix environments, because they have relative tolerance to hydrolysis and / or enzyme-mediated degradation. Compared with other related compounds, the compounds of the present invention show improved solution stability.

[0229] In one aspect, the present invention provides a compound having formula (I):

[0230] AYB

[0231] (I)

[0232] Where A is:

[0233]

[0234] in

[0235] b and w are each independently an integer from 0 to 7, and v is an integer from 0 to 5, provided that:

[0236] The sum of b, v, and w is at least 3; and

[0237] The sum of b and w ranges from 0 to 7;

[0238] z is 1 or 2;

[0239] X is selected from -S-, -S(=O)- and -S(=O)2-;

[0240] Z1 and Z2 are each independently selected from the group consisting of: -O-, -NR-, -S-, S(=O), -S(=O)2-, -C(=O)O-, -OC(=O)-, -C(=O)NR-, -NRC(=O)-, -C(=O)S-, -SC(=O)-, -OC(=O)O-, NRC(=O)O-, -OC(=O)NR-, and -NRC(=O)NR-;

[0241] R 11 、R 12 、R x 、R y 、R 14 、R 15 、R 16 and R 17 Each independently is H or a C1-C6 aliphatic group;

[0242] R, R 13 and R 18 Each independently is H or a C1-C6 aliphatic group;

[0243] R 19 is H, a C1-C6 aliphatic group, an amino protecting group, L3-C(=O)-, or A2;

[0244] L1 and L2 are each independently C5-C 21 Aliphatic or C4-C 20 heteroaliphatic groups;

[0245] L3 is C1-C 21 Aliphatic or C2-C 20 heteroaliphatic groups;

[0246] A2 is an amino acid or peptide;

[0247] Among them, there are 11 、R 12 、R 13 、R 14 、R 15 、R 16 、R 17 、R 18 、R 19 、R x 、R y Any aliphatic or heteroaliphatic group in any of L1, L2, and L3 is optionally substituted;

[0248] Y is

[0249]

[0250] wherein R1 and R2 are independently selected from the group consisting of H, -CH2OH, -CH2CH2OH, -CH(CH3)OH, -CH2OPO(OH)2, -CH2C(=O)NH2, -CH2CH2C(=O)OH, and -CH2CH2C(=O)OR8, wherein any of the alkyl hydrogens may be substituted by halogen;

[0251] R8 is selected from the group consisting of H and linear or branched C1-C6 alkyl;

[0252] and

[0253] B comprises polyethylene glycol (PEG) or consists of polyethylene glycol (PEG),

[0254] or a pharmaceutically acceptable salt, solvate or prodrug thereof.

[0255] In some embodiments, v is an integer selected from 2, 3, 4, or 5. In some embodiments, v is 2.

[0256] In some embodiments, R x 、R y 、R 11 、R 12 、R 13 、R 14 、R 15 、R 16 and R 17 It’s H.

[0257] In some embodiments, Z1 and Z2 are the same and are selected from the group consisting of: -O-, -NR-, -S-, S(=O), S(=O)2-, -C(=O)O-, -OC(=O)-, -C(=O)NR-, -NRC(=O)-, -C(=O)S-, -SC(=O)-, OC(=O)O-, NRC(=O)O-, -OC(=O)NR-, and -NRC(=O)NR-.

[0258] In some embodiments, Z1 and Z2 are independently selected from the group consisting of: -C(=O)O-, -OC(=O)-, -C(=O)NR-, -NRC(=O)-, -C(=O)S-, -SC(=O)-, -OC(=O)O-, -NRC(=O)O-, -OC(=O)NR-, and -NRC(=O)NR-.

[0259] In some embodiments, w is an integer selected from 1 to 7. In some embodiments, w is 1.

[0260] In some embodiments, b is 0.

[0261] In some embodiments, R 19 Selected from the group consisting of: H, C1-C6 alkyl, -C(=O)C1-C6 alkyl, or -C(=O)C 11 -C 19 alkyl.

[0262] In some embodiments, L1 and L2 are independently selected from C5-C 21 Aliphatic or C4-C 20 In some embodiments, L1 and L2 are independently selected from C 10 -C 18 Aliphatic or C 10 -C 18 In some embodiments, L1 and L2 are independently selected from C 14 -alkyl and C 15 -alkyl. In some embodiments, L1 and L2 are branched C 5-21 Aliphatic group. Branched C 5-21 The aliphatic radical may be branched at the carbon atom to which Z1 or Z2 is bonded.

[0263] In some embodiments, the present invention provides a compound having formula (I), wherein

[0264] v is an integer selected from 2 to 5;

[0265] b is 0;

[0266] R x 、Ry 、R 13 、R 14 、R 15 、R 16 and R 17 It is H;

[0267] Z1 and Z2 are independently selected from the group consisting of: -C(=O)O-, -OC(=O)-, -C(=O)NR-, -NRC(=O)-, -C(=O)S-, -SC(=O)-, -OC(=O)O-, -NRC(=O)O-, -OC(=O)NR-, and -NRC(=O)NR-;

[0268] w is an integer selected from 1 to 7;

[0269] R 19 Selected from the group consisting of: H, C1-C6 alkyl, -C(=O)C1-C6 alkyl, or -C(=O)C 11 -C 19 alkyl; and

[0270] L1 and L2 are independently selected from C 10 -C 18 Aliphatic or C 10 -C 18 Heteroaliphatic group.

[0271] In some embodiments, X is S.

[0272] In some embodiments, X is S(=O).

[0273] In some embodiments, X is S(=O)2.

[0274] In some embodiments, B indicates a substituted PEG.

[0275] In some embodiments, B is a substituted PEG according to the following formula BI:

[0276]

[0277] in

[0278] n is 3 to 100;

[0279] m is 1, 2, 3 or 4;

[0280] p is 2, 3, or 4;

[0281] q is zero or 1;

[0282] R3 is H, -NH2 or -OH, wherein when q is zero, R3 is H, and when q is 1, R3 is -NH2 or -OH;

[0283] L is zero or consists of 1 to 10 units, each of which is a natural alpha amino acid or derived from a natural alpha amino acid and has the formula:

[0284]

[0285] wherein R4 is H; and

[0286] R5 is the side chain, or the second hydrogen of the amino acid.

[0287] In some embodiments, B is a substituted PEG according to the following formula B-II:

[0288]

[0289] in

[0290] p is 2, 3, or 4;

[0291] n is 3 to 100;

[0292] m is 1, 2, 3 or 4;

[0293] t is 2, 3, or 4;

[0294] k is 3 to 100;

[0295] h is 1, 2, 3, or 4;

[0296] q is zero or 1;

[0297] wherein when q is 1, R3 is -NH2 or -OH;

[0298] Wherein when q is zero, R3 is H;

[0299] L is zero or consists of 1 to 10 units, each of which is a natural alpha amino acid or derived from a natural alpha amino acid and has the formula:

[0300]

[0301] wherein R4 is H; and

[0302] R5 is the side chain, or the second hydrogen of the amino acid.

[0303] In some embodiments of the substituted PEG having formula BI or B-II, q is 1.

[0304] In some embodiments of substituted PEGs having formula BI or B-II, n can be from 10 to 14, such as 11, or from 24 to 30, such as 27.

[0305] In some embodiments of substituted PEGs having formula BI or B-II, m is from 1 to 3, such as 2.

[0306] In some embodiments of the substituted PEG having formula BI or B-II, when q is 1, R3 is -NH2.

[0307] In some embodiments of the substituted PEG having formula BI or B-II, L is a natural alpha amino acid residue.

[0308] In another aspect, the present invention provides a compound comprising moiety (G) according to the formula:

[0309]

[0310] wherein R1 and R2 are independently selected from the group consisting of H, -CH2OH, -CH2CH2OH, -CH(CH3)OH, -CH2OPO(OH)2, -CH2C(=O)NH2, -CH2CH2C(=O)OH, and -CH2CH2C(=O)OR8, wherein any of the alkyl hydrogens may be substituted by halogen;

[0311] R8 is selected from the group consisting of H and linear or branched C1-C6 alkyl;

[0312] Z1, L1, Z2, L2, X, b, w, v, z, R x 、R y 、R 11 、R 12 、R 13 、R 14 、R 15 、R 16 、R 17 、R 18 and R 19 is as defined above for the compound of formula (I);

[0313] The above structure is covalently linked to polyethylene glycol (PEG),

[0314] or a pharmaceutically acceptable salt, solvate or prodrug thereof.

[0315] In some embodiments, moieties G and PEG are expressed by Typically, the PEG is substituted and is linked at one end by an amide linker (including covalently linked to the indicated covalently linked carbonyl).

[0316] In some embodiments, PEG is a substituted PEG. In some embodiments, the substituted PEG is indicated by the formula:

[0317]

[0318] in

[0319] n is 3 to 100;

[0320] m is 1, 2, 3 or 4;

[0321] p is 2, 3, or 4;

[0322] q is zero or 1;

[0323] R3 is H, -NH2 or -OH, wherein when q is zero, R3 is H, and when q is 1, R3 is -NH2 or -OH;

[0324] L is zero or consists of 1 to 10 units, each of which is a natural alpha amino acid or derived from a natural alpha amino acid and has the formula:

[0325]

[0326] wherein R4 is H; and

[0327] R5 is the side chain, or the second hydrogen of the amino acid.

[0328] In some embodiments, when q is 1, R3 is -NH2.

[0329] In some embodiments, L is a natural alpha amino acid residue.

[0330] In one aspect, the present invention provides a compound having formula (VI):

[0331]

[0332] in

[0333] n is 3 to 100;

[0334] m is 1, 2, 3 or 4;

[0335] p is 2, 3, or 4;

[0336] q is zero or 1;

[0337] R1 and R2 are independently selected from the group consisting of H, -CH2OH, -CH2CH2OH, -CH(CH3)OH, -CH2OPO(OH)2, -CH2C(=O)NH2, -CH2CH2C(=O)OH, and -CH2CH2C(=O)OR8, wherein any of the alkyl hydrogens may be substituted by halogen;

[0338] R8 is selected from the group consisting of H and linear or branched C1-C6 alkyl;

[0339] wherein when q=1, R3 is -NH2 or -OH;

[0340] Wherein when q=0, R3 is H;

[0341] L is zero or consists of 1 to 10 units, each of which is a natural alpha amino acid or derived from a natural alpha amino acid and has the formula:

[0342]

[0343] wherein R4 is H; and

[0344] R5 is the side chain, or the second hydrogen of the amino acid,

[0345] b and w are each independently an integer from 0 to 7, and v is an integer from 0 to 5, provided that:

[0346] The sum of b, v, and w is at least 3; and

[0347] The sum of b and w ranges from 0 to 7;

[0348] z is 1 or 2;

[0349] X is selected from -S-, -S(=O)- and -S(=O)2-;

[0350] Z1 and Z2 are each independently selected from the group consisting of: -O-, -NR-, -S-, -S(=O)-, -S(=O)2-, -C(=O)O-, -OC(=O)-, -C(=O)NR-, -NRC(=O)-, -C(=O)S-, -SC(=O)-, -OC(=O)O-, -NRC(=O)O-, -OC(=O)NR-, and -NRC(=O)NR-;

[0351] In each case of b, v, w, and z, R 11 、R 12 、R x 、R y 、R 14 、R 15 、R 16 and R 17 Each independently is H or a C1-C6 aliphatic group;

[0352] R, R 13 and R 18 Each independently is H or a C1-C6 aliphatic group;

[0353] R 19 is H, a C1-C6 aliphatic group, an amino protecting group, L3-C(=O)-, or A2;

[0354] L1 and L2 are each independently C5-C 21 Aliphatic or C4-C 20 heteroaliphatic groups;

[0355] L3 is C1-C 21 Aliphatic or C2-C 20 heteroaliphatic groups;

[0356] A2 is an amino acid or peptide;

[0357] Among them, there are 11 、R 12 、R 13 、R 14 、R 15 、R 16 、R 17 、R 18 、R 19 、R x 、R y Any aliphatic or heteroaliphatic group in any of L1, L2, and L3 is optionally substituted;

[0358] or a pharmaceutically acceptable salt, solvate or prodrug thereof.

[0359] In some embodiments of compounds having formula (VI), v is 2, 3, 4, or 5, preferably 2 or 3, most preferably 2.

[0360] In some embodiments of compounds of Formula (VI), b is 0.

[0361] In some embodiments of compounds of Formula (VI), z is 1.

[0362] In some embodiments, compounds having Formula (VI) are represented by Formula (VI'):

[0363]

[0364] Among them, Z1, L1, Z2, L2, X, b, w, z, R 11 、R 12 、R 13 、R 14 、R 15 、R 16 、R 17 、R 18 and R 19 is as defined in the compound of formula (I) above;

[0365] R1, R2, p, n, m, L, q and R3 are as defined in the compound having formula (VI) above;

[0366] v" is 1, 2, 3 or 4, preferably 1;

[0367] R j and R k are independently selected from H or C1-C6 aliphatic, preferably H; and

[0368] R in each case x” and R y” Independently selected from H or C1-C6 aliphatic, preferably H;

[0369] or a pharmaceutically acceptable salt, solvate or prodrug thereof.

[0370] In some embodiments, the compound is a compound having Formula (VII):

[0371] AY-NH-(CH2) p -O-(CH2-CH2-O) n -[(CH2) m -CO-L-] q R3

[0372] (VII)

[0373] wherein A and Y are as defined for the compound of formula (I), and

[0374] n is 3 to 100;

[0375] m is 1, 2, 3 or 4;

[0376] p is 2, 3, or 4;

[0377] q is zero or 1;

[0378] wherein when q is 1, R3 is -NH2 or -OH;

[0379] Wherein when q is zero, R3 is H;

[0380] L is zero or consists of 1 to 10 units, each of which is a natural alpha amino acid residue or is derived from a natural alpha amino acid and has the formula:

[0381]

[0382] wherein R4 is H; and

[0383] R5 is the side chain, or the second hydrogen of the amino acid,

[0384] or a pharmaceutically acceptable salt, solvate or prodrug thereof.

[0385] In some embodiments, the compound is a compound having Formula (X):

[0386]

[0387] in

[0388] n is 3 to 100;

[0389] k is 3 to 100;

[0390] m is 1, 2, 3 or 4;

[0391] p is 2, 3, or 4;

[0392] t is 2, 3, or 4;

[0393] h is 1, 2, 3, or 4;

[0394] q is zero or 1;

[0395] wherein R1 and R2 are independently selected from the group consisting of H, -CH2OH, -CH2CH2OH, -CH(CH3)OH, -CH2OPO(OH)2, -CH2C(=O)NH2, -CH2CH2C(=O)OH, and -CH2CH2C(=O)OR8, wherein any of the alkyl hydrogens may be replaced by halogen;

[0396] R8 is selected from the group consisting of H and linear or branched C1-C6 alkyl;

[0397] wherein when q=1, R3 is -NH2 or -OH;

[0398] Wherein when q=0, R3 is H;

[0399] L is zero or consists of 1 to 10 units, each of which is a natural alpha amino acid or derived from a natural alpha amino acid and has the formula:

[0400]

[0401] wherein R4 is H; and

[0402] R5 is the side chain, or the second hydrogen of the amino acid,

[0403] b and w are each independently an integer from 0 to 7, and v is an integer from 0 to 5, provided that:

[0404] The sum of b, v, and w is at least 3; and

[0405] The sum of b and w ranges from 0 to 7;

[0406] z is 1 or 2;

[0407] X is selected from -S-, -S(=O)- and -S(=O)2-;

[0408] Z1 and Z2 are each independently selected from the group consisting of: -O-, -NR-, -S-, -S(=O)-, -S(=O)2-, -C(=O)O-, -OC(=O)-, -C(=O)NR-, -NRC(=O)-, -C(=O)S-, -SC(=O)-, -OC(=O)O-, -NRC(=O)O-, -OC(=O)NR-, and -NRC(=O)NR-;

[0409] In each case of b, v, w, and z, R 11 、R 12 、R x 、R y 、R 14 、R 15 、R 16 and R 17 Each independently is H or a C1-C6 aliphatic group;

[0410] R, R 13 and R 18 Each independently is H or a C1-C6 aliphatic group;

[0411] R 19 is H, a C1-C6 aliphatic group, an amino protecting group, L3-C(=O)-, or A2;

[0412] L1 and L2 are each independently C5-C 21 Aliphatic or C4-C 20 heteroaliphatic groups;

[0413] L3 is C1-C 21 Aliphatic or C2-C 20 heteroaliphatic groups;

[0414] A2 is an amino acid or peptide;

[0415] Among them, there are 11 、R 12 、R 13 、R 14 、R 15 、R 16 、R 17 、R 18 、R 19 、R x 、R y Any aliphatic or heteroaliphatic group in any of L1, L2, and L3 is optionally substituted;

[0416] or a pharmaceutically acceptable salt, solvate or prodrug thereof.

[0417] The present invention also provides compositions comprising a compound of the present invention or a pharmaceutically acceptable salt, solvate or prodrug thereof, and a pharmaceutically acceptable carrier, diluent or excipient. Any of the compounds described herein or variants thereof may be included in the compositions of the present invention.

[0418] As discussed above, the present invention provides Toll-like receptor 2 protein (TLR2) agonist compounds and compositions thereof. In the human body, TLR2 plays a fundamental role in recognizing pathogens and activating the innate immune response. It is encoded by the TLR2 gene and expressed on the surface of specific cells.

[0419] Without wishing to be bound by any theory or mode of action, it is believed that the compounds of this invention as described herein are agonists of TLR2, and are active by combining and stimulating the innate immune system at TLR2. The innate immune system forms an immediate defense against pathogens, such as cells that infect the respiratory tract and replicate therein. Studies have shown that the medicaments that stimulate the innate immune system can be used to limit respiratory infections, and these medicaments can provide protection in isolation, and can provide protection during the period between inoculation and antibody and immune cell formation, to avoid infection. Such medicaments are considered to be useful for treating and / or preventing respiratory infections in a non-antigen-specific manner, or respiratory conditions (such as influenza A, pneumonia) associated with infectious agents such as viruses or bacteria or by infectious agents.

[0420] In this regard, the compounds of the invention as described herein may have activity in both activating human TLR2 and inhibiting viral progression that is at least comparable to other TLR2 agonists such as Pam2Cys-Ser-K4, Pam2Cys-Ser-Ser-PEG, and Pam3Cys-Ser-PEG.

[0421] As used herein, 'Ser' refers to the amino acid serine, and 'Cys' refers to the amino acid cysteine.

[0422] As used herein, 'PEG' refers to the polymer compound polyethylene glycol. Unless otherwise defined, references to 'PEG' include ethylene oxide polymers of any length. References to PEG also include substituted PEGs. In some embodiments, substituted PEGs can be defined by Formula BI or B-II as described herein.

[0423] Thus, in one aspect, the present invention provides a method for treating and / or preventing a disease, comprising enhancing the innate immune response in a subject by administering to a subject in need thereof an effective amount of a compound of the present invention as described herein, or a pharmaceutically acceptable salt, solvate or prodrug thereof.

[0424] In another aspect, the present invention provides a method for treating and / or preventing a disease caused by an infectious agent, comprising administering to a subject in need thereof an effective amount of a compound of the present invention as described herein or a pharmaceutically acceptable salt, solvate or prodrug thereof.

[0425] In another aspect, the present invention provides a method for treating and / or preventing respiratory diseases or conditions associated with viral or bacterial infections, comprising administering to a subject in need thereof a compound of the present invention as described herein, or a pharmaceutically acceptable salt, solvate or prodrug thereof.

[0426] In another aspect, the present invention provides a method for treating and / or preventing respiratory tract infection, comprising administering to a subject in need thereof a compound of the present invention as described herein or a pharmaceutically acceptable salt, solvate or prodrug thereof. Preferably, the method further comprises the step of identifying a subject suffering from a respiratory tract infection.

[0427] In another aspect, the present invention provides a method for reducing airway inflammation, comprising administering to a subject in need thereof a compound of the present invention as described herein, or a pharmaceutically acceptable salt, solvate, or prodrug thereof.

[0428] The present invention also provides a method for improving a subject's ability to control a respiratory disease or condition during a respiratory viral infection, the method comprising administering to a subject in need thereof a compound of the present invention as described herein, or a pharmaceutically acceptable salt, solvate, or prodrug thereof. Preferably, the infection is not a rhinovirus infection.

[0429] The present invention also provides a method for treating and / or preventing diseases or conditions associated with TLR2 receptor, comprising administering to a subject in need thereof a compound of the present invention as described herein or a pharmaceutically acceptable salt, solvate or prodrug thereof.

[0430] The present invention also provides a method for stimulating TLR2 activity in a cell, comprising contacting the cell with a compound of the present invention as described herein, or a pharmaceutically acceptable salt, solvate, or prodrug thereof. In some embodiments, the cell is contacted with the compound by administering the compound, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, or a composition comprising the compound, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, to a subject in need thereof. In some embodiments, the cell is provided in the form of a cell culture.

[0431] In another aspect, the present invention provides use of a compound of the present invention as described herein, or a pharmaceutically acceptable salt, solvate or prodrug thereof, in the preparation of a medicament for enhancing an innate immune response in a subject.

[0432] In another aspect, the present invention provides the use of a compound of the present invention as described herein, or a pharmaceutically acceptable salt, solvate or prodrug thereof, in the preparation of a medicament for treating and / or preventing a disease caused by an infectious agent.

[0433] In another aspect, the present invention further provides the use of a compound of the present invention as described herein, or a pharmaceutically acceptable salt, solvate or prodrug thereof, in the preparation of a medicament for treating and / or preventing a respiratory disease or condition associated with a viral or bacterial infection in a subject.

[0434] In another aspect, the present invention further provides use of a compound of the present invention as described herein, or a pharmaceutically acceptable salt, solvate or prodrug thereof, in the preparation of a medicament for treating and / or preventing a respiratory tract infection in a subject.

[0435] In yet another aspect, the present invention provides use of a compound of the present invention as described herein, or a pharmaceutically acceptable salt, solvate or prodrug thereof, in the preparation of a medicament for treating and / or preventing respiratory tract infections.

[0436] In another aspect, the present invention further provides use of a compound of the present invention as described herein, or a pharmaceutically acceptable salt, solvate or prodrug thereof, in the preparation of a medicament for reducing airway inflammation.

[0437] In another aspect, the present invention further provides the use of a compound of the present invention as described herein, or a pharmaceutically acceptable salt, solvate or prodrug thereof, in the preparation of a medicament for improving a subject's ability to control a respiratory disease or condition during a respiratory viral infection. Preferably, the infection is not a rhinovirus infection.

[0438] In another aspect, the present invention further provides use of a compound of the present invention as described herein, or a pharmaceutically acceptable salt, solvate or prodrug thereof, in the preparation of a medicament for treating and / or preventing a disease or condition associated with the TLR2 receptor.

[0439] In one aspect, the present invention provides the use of a compound of the present invention as described herein, or a pharmaceutically acceptable salt, solvate or prodrug thereof, for enhancing an innate immune response in a subject.

[0440] In another aspect, the present invention provides the use of a compound of the present invention as described herein, or a pharmaceutically acceptable salt, solvate or prodrug thereof, for preventing a disease caused by an infectious agent in a subject.

[0441] In another aspect, the present invention provides the use of a compound of the present invention as described herein, or a pharmaceutically acceptable salt, solvate or prodrug thereof, for treating and / or preventing a respiratory disease or disorder associated with a viral or bacterial infection in a subject.

[0442] In another aspect, the present invention provides the following uses of a compound of the present invention as described herein, or a pharmaceutically acceptable salt, solvate or prodrug thereof: for (a) treating and / or preventing a respiratory tract infection in a subject; (b) reducing airway inflammation in a subject; (c) controlling a respiratory tract disease or condition during a respiratory viral infection in a subject; (d) for treating and / or preventing a disease or condition associated with a TLR2 receptor.

[0443] In any of these aspects, the compound can be administered in the form of a composition. Typically, the composition further comprises a pharmaceutically acceptable carrier, diluent, or excipient. The composition can be formulated for administration to the upper and / or lower respiratory tract, for example, by inhalation or intranasally.

[0444] In any aspect of the invention, the compounds of the invention as described herein, or pharmaceutically acceptable salts, solvates or prodrugs thereof, may be conjugated to other compounds. The other compound is any of those described herein.

[0445] In any aspect of the invention, the compound of the invention as described herein, or a pharmaceutically acceptable salt, solvate or prodrug thereof, is administered once daily or once weekly.

[0446] In any aspect of the invention where prevention or prophylaxis is intended or required, the compound is administered to the subject prior to the appearance of any clinically or biochemically detectable symptoms of viral infection.

[0447] In any aspect of the invention, administration of a compound of the invention as described herein, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, to a subject reduces viral load in the subject. Preferably, the viral load is reduced in the respiratory tract, e.g., the upper and / or lower respiratory tract. Preferably, the viral load is reduced in the lungs.

[0448] In any aspect herein, the infectious agent can be a virus. Preferably, the virus is a virus associated with respiratory tract infection. Even more preferably, the virus is an influenza virus. In any aspect, the virus is not a rhinovirus.

[0449] Influenza (commonly known as "flu") is an infectious disease affecting birds and mammals caused by RNA viruses (influenza viruses) of the Orthomyxoviridae family. The most common symptoms of the disease are chills, fever, sore throat, muscle aches, severe headache, cough, weakness / fatigue, and a general feeling of being unwell.

[0450] Influenza viruses make up three of the five genera of the Orthomyxoviridae family. Influenza A and B viruses co-circulate during seasonal epidemics and can cause severe influenza infections. Influenza C virus infections are less common but can be severe and cause localized epidemics.

[0451] Influenza A viruses can be subdivided into different serotypes, or subtypes, based on the antibody response to these viruses. Influenza A viruses are divided into multiple subtypes based on two proteins on the surface of the virus: hemagglutinin (H) and neuraminidase (N). There are 18 different hemagglutinin subtypes and 11 different neuraminidase subtypes (H1 to H18 and N1 to N11, respectively). The subtypes recognized in humans are H1N1, H1N2, H2N2, H3N2, H5N1, H7N2, H7N3, H7N7, H9N2, and H10N7.

[0452] Influenza has a significant impact on public health, with severe economic consequences in addition to devastating health problems including morbidity and even mortality. Therefore, there is a need for therapeutic agents that can prevent infection or reduce the severity of infection in individuals.

[0453] In any aspect or embodiment of the invention, the influenza infection to be treated or prevented is a viral infection selected from the group consisting of influenza A, B, or C.

[0454] The term 'respiratory disease' or 'respiratory condition' refers to any of several diseases that involve inflammation and affect components of the respiratory system, including the upper respiratory tract (including the nasal cavity, pharynx, and larynx) and the lower respiratory tract (including the trachea, bronchi, and lungs). Inflammation in the upper and lower respiratory tracts may be associated with or caused by viral infections or allergens. It is expected that the anti-inflammatory activity of the compounds, when administered alone or in combination with glucocorticoids, will make them particularly suitable for treating these diseases or conditions.

[0455] The symptoms of respiratory diseases may include cough, excessive sputum, dyspnea or chest tightness, with audible wheezing. Exercise capacity may be greatly limited. In asthma, the FEV1.0 (forced expiratory volume in one second) as a percentage of the value predicted by the nomogram based on weight, height and age may reduce because peak expiratory flow rate may be present during forced exhalation. In COPD, the FEV1.0 as the ratio of FVC is typically reduced to less than 0.7. The impact of each of these diseases can also be measured by not going to work / school days, sleep disturbances, the needs of bronchodilator drugs, the needs of glucocorticoids (including oral glucocorticoids).

[0456] The presence, improvement, treatment or prevention of respiratory disease can be determined by any clinical or biochemical method of the subject or a biopsy thereof. For example, the parameters measured can be the presence or degree of lung function, signs and symptoms of obstruction; exercise tolerance; nighttime awakenings; number of days missed from school or work; use of bronchodilators; dose of inhaled corticosteroids (ICS); use of oral glucocorticoids (GC); need for other medications; need for medical treatment; hospital admission.

[0457] As used herein, the term respiratory tract infection means an infection anywhere in the respiratory tract caused by a virus or bacteria. Examples of respiratory tract infections include, but are not limited to, a cold, sinusitis, throat infection, tonsillitis, laryngitis, bronchitis, pneumonia, or bronchiolitis. Preferably, in any embodiment of the present invention, the respiratory tract infection is a cold.

[0458] Individuality can be determined as suffering from respiratory tract infection by viral testing, and may show symptoms of itchy eyes, runny nose, nasal congestion, sneezing, sore throat, cough, headache, fever, discomfort, fatigue and weakness. On the one hand, the subject suffering from respiratory tract infection may not have any other respiratory tract disease. The presence or amount of detection virus can be carried out by PCR / sequencing or by serology to the RNA separated from clinical samples (nasal wash, sputum, BAL).

[0459] The term "pharmaceutically acceptable" may be used to describe any pharmaceutically acceptable salt, hydrate or prodrug, or any other compound that, when administered to a subject, is capable of (directly or indirectly) providing a compound of the invention as described herein, or a pharmaceutically acceptable salt, solvate or prodrug thereof, or an active metabolite or residue thereof.

[0460] Suitable pharmaceutically acceptable salts may include, but are not limited to, salts of pharmaceutically acceptable inorganic acids such as hydrochloric acid, sulfuric acid, phosphoric acid, nitric acid, carbonic acid, boric acid, sulfamic acid, and hydrobromic acid; or salts of pharmaceutically acceptable organic acids such as acetic acid, propionic acid, butyric acid, tartaric acid, maleic acid, hydroxymaleic acid, fumaric acid, malic acid, citric acid, lactic acid, mucic acid, gluconic acid, benzoic acid, succinic acid, oxalic acid, phenylacetic acid, methanesulfonic acid, toluenesulfonic acid, benzenesulfonic acid, salicylic acid, sulfanilic acid, aspartic acid, glutamic acid, edetic acid, stearic acid, palmitic acid, oleic acid, lauric acid, pantothenic acid, tannic acid, ascorbic acid, and valeric acid.

[0461] Basic salts may include, but are not limited to, those formed with pharmaceutically acceptable cations such as sodium, potassium, lithium, calcium, magnesium, zinc, ammonium, alkylammonium (such as salts formed by triethylamine), alkoxyammonium (such as those formed with ethanolamine), and salts formed by ethylenediamine, choline, or amino acids such as arginine, lysine, or histidine. General information about the types of pharmaceutically acceptable salts and their formation is known to those skilled in the art and as described in general texts such as "Handbook of Pharmaceutical Salts," PHS Tahl, CG Wermuth, 1st edition, 2002, Wiley-VCH.

[0462] Where the compounds are solids, those skilled in the art will appreciate that the compounds, agents, solvates and salts of the invention may exist in different crystalline or polymorphic forms, all of which are intended to be within the scope of the invention and the specified formula.

[0463] The term "polymorph" includes any crystalline form of the compounds of the present invention as described herein, such as anhydrous forms, hydrous forms, solvate forms, and mixed solvate forms.

[0464] It should be understood that the compounds of the present invention can have chiral centers and therefore can exist in R-configuration or S-configuration. The compound can be provided as a racemate or in the form of enantiomer or diastereomer enrichment. The enantiomer and diastereomer enrichment of the compound can be obtained by asymmetric synthesis, incorporating chiral pool materials, or by stereoselective separation. Therefore, the compound can be provided as a purified enantiomer or diastereomer or as a mixture of any ratio. Isomers can be separated conventionally by chromatography or using a resolving agent. Alternatively, chiral intermediates can be used to prepare independent isomers by asymmetric synthesis. When the compound has a carbon-carbon double bond, it can occur in the form of Z- or E- and all isomeric forms of the compound are included in the present invention.

[0465] The compounds of the present invention are intended to include solvated and unsolvated forms of the compounds, where applicable. As used herein, the term "solvate" refers to a complex having a variable stoichiometry formed by combining a solvent with a compound of the present invention. Thus, a solvate may contain a substoichiometric amount of solvent, an equimolar amount of solvent, or a stoichiometric excess of solvent relative to the compound of the present invention. Such solvents for the purposes of the present invention may not interfere with the biological activity of the solute. Examples of suitable solvents include, but are not limited to, water, methanol, ethanol, and acetic acid. Preferably, the solvent used is a pharmaceutically acceptable solvent. Examples of suitable pharmaceutically acceptable solvents include, but are not limited to, water, ethanol, and acetic acid. Most preferably, the solvent used is water. A solvate wherein the solvent is water may be referred to as a hydrate.

[0466] Basic nitrogen-containing groups can be quaternized with reagents such as lower alkyl halides, such as methyl, ethyl, propyl, and butyl chlorides, bromides, and iodides; dialkyl sulfates, such as dimethyl sulfate and diethyl sulfate; and the like.

[0467] Compounds as described herein also include isotopic variations, such as the substitution of deuterium for hydrogen.

[0468] The compounds of the present invention can exist and be separated in optically active forms and racemic forms. As will be appreciated by those skilled in the art, the present invention is intended to encompass any racemic, optically active or stereoisomeric forms of the compounds of the present invention having the useful properties described herein, or mixtures thereof. Methods for preparing these forms (e.g., by resolving the racemic mixture by recrystallization, by synthesizing from optically active starting materials, by chiral synthesis, or by chiral chromatographic separation) are well known in the art. In a preferred embodiment, for the carbons shown below with *, the compounds of the present invention are provided as racemic mixtures. In another preferred aspect, the compounds of the present invention comprise

[0469]

[0470] The structure has an excess of or only L-configuration or naturally occurring amino acids.

[0471] In any aspect or embodiment of the invention, the compounds of the invention may be provided in a chiral form enriched at the chiral center at the carbon atom (shown at *) in the following moiety A:

[0472]

[0473] wherein the chiral center is in the R configuration. In some embodiments, this stereoisomer of the compound can be depicted as:

[0474] Among them, L1, L2, Z1, Z2, R x 、R y 、R 11 、R 12 、R 13 、R 14 、R 15 、R 16 、R 17 、R 18 、R 19 , b, v and z are as defined for the compound of formula (I), and w is 1. Other stereocenters in these compounds may be racemic or enriched in the R or S configuration.

[0475] In any aspect or embodiment of the invention, the compounds of the invention may be provided in a chiral form enriched at the chiral center at the carbon atom (shown at *) in the following moiety A:

[0476]

[0477] wherein the chiral center is in the S configuration. In some embodiments, portion A of this stereoisomer of the compound can be depicted as:

[0478] Among them, L1, L2, Z1, Z2, R x 、R y 、R 11 、R 12 、R 13 、R 14 、R 15 、R 16 、R 17 、R 18 、R 19 , b, v, w and z are as defined for compounds of formula (I). Other stereocenters in these compounds may be racemic or enriched in the R or S configuration.

[0479] In any aspect or embodiment of the invention, the compounds of the invention may be provided in a chiral form enriched at the chiral center at the carbon atom (shown at **) in the following moiety A:

[0480]

[0481] Wherein the chiral center is in L configuration. This form of compound may also be referred to as the L-Cys analog stereoisomer of the compound of the present invention. In some embodiments, this stereoisomer of the compound can be depicted as:

[0482] Among them, L1, L2, Z1, Z2, R x 、R y 、R 11 、R 12 、R 13 、R 14 、R 15 、R 16 、R 17 、R 18 、R 19 , b, v, w and z are as defined for compounds of formula (I). Other stereocenters in these compounds may be racemic or enriched in the R or S configuration.

[0483] In any aspect or embodiment of the invention, the compounds of the invention may be provided in a chiral form enriched at the chiral center at the carbon atom (shown at **) in the following moiety A:

[0484]

[0485] Wherein the chiral center is in the D configuration. This form of compound may also be referred to as the D-Cys analog stereoisomer of the compound of the present invention. In some embodiments, the portion A of this stereoisomer of the compound may be depicted as:

[0486] Among them, L1, L2, Z1, Z2, R x 、R y 、R 11 、R 12 、R 13 、R 14 、R 15 、R 16 、R 17 、R 18 、R 19 , b, v and z are as defined for the compound of formula (I), and w is 1. Other stereocenters in these compounds may be racemic or enriched in the R or S configuration.

[0487] In any aspect or embodiment of the invention, the compounds of the invention may be provided in a chiral form enriched at the chiral center at the carbon atom (shown at * and **) in the following moiety A:

[0488]

[0489] wherein the chiral center * is in the R configuration and the chiral center ** is in the R configuration. This form of the compound may also be referred to as an R,R-Cys analog stereoisomer of the compound of the present invention. In some embodiments, this stereoisomer of the compound can be depicted as:

[0490] Among them, L1, L2, Z1, Z2, R x 、R y 、R 11 、R 12 、R 13 、R 14 、R 15 、R 16 、R 17 、R 18 and R 19 is as defined for compounds having formula (I), b is 0, v is 1, z is 1, and w is 1. Other stereocenters in these compounds may be racemic or enriched in the R or S configuration.

[0491] In any aspect or embodiment of the invention, the compounds of the invention may be provided in a chiral form enriched at the chiral center at the carbon atom (shown at * and **) in the following moiety A:

[0492]

[0493] wherein the chiral center * is in the S configuration and the chiral center ** is in the R configuration. This form of the compound may also be referred to as an S,R-Cys analog stereoisomer of the compound of the present invention. In some embodiments, portion A of this stereoisomer of the compound may be depicted as:

[0494] Among them, L1, L2, Z1, Z2, R x 、R y 、R 11 、R 12 、R 13 、R 14 、R 15 、R 16 、R 17 、R 18 and R 19 is as defined for compounds having formula (I), b is 0, v is 1, z is 1, and w is 1. Other stereocenters in these compounds may be racemic or enriched in the R or S configuration.

[0495] In any aspect or embodiment of the invention, the compounds of the invention may be provided in a chiral form enriched at the chiral center at the carbon atom (shown at * and **) in the following moiety A:

[0496]

[0497] wherein the chiral center * is in the S configuration and the chiral center ** is in the S configuration. This form of the compound may also be referred to as the S,S-Cys analog stereoisomer of the compound of the present invention. In some embodiments, portion A of this stereoisomer of the compound may be depicted as:

[0498] Among them, L1, L2, Z1, Z2, R x 、R y 、R 11 、R 12 、R 13 、R 14 、R 15 、R 16 、R 17 、R 18 and R 19 is as defined for compounds having formula (I), b is 0, v is 1, z is 1, and w is 1. Other stereocenters in these compounds may be racemic or enriched in the R or S configuration.

[0499] In any aspect or embodiment of the invention, the compounds of the invention may be provided in a chiral form enriched at the chiral center at the carbon atom (shown at * and **) in the following moiety A:

[0500]

[0501] wherein the chiral center * is in the R configuration and the chiral center ** is in the S configuration. This form of the compound may also be referred to as an R,S-Cys analog stereoisomer of the compound of the present invention. In some embodiments, portion A of this stereoisomer of the compound may be depicted as:

[0502] Among them, L1, L2, Z1, Z2, R x 、R y 、R 11 、R 12 、R 13 、R 14 、R 15 、R 16 、R 17 、R 18 and R 19 is as defined for compounds having formula (I), b is 0, v is 1, z is 1, and w is 1. Other stereocenters in these compounds may be racemic or enriched in the R or S configuration.

[0503] In any aspect or embodiment of the invention, the compounds of the invention comprise a chiral center (shown at ***) in the Y portion of the compound:

[0504]

[0505] Wherein the chiral center is in the L-configuration. This form of the compound may also be referred to as the LY stereoisomer of the compound of the present invention. The stereochemistry of the chiral center in the LY stereoisomer can be combined with other chiral centers of the compound (such as the chiral center in part A described herein) without restriction.

[0506] In any aspect or embodiment of the invention, the compounds of the invention comprise a chiral center (shown at ***) in the Y portion of the compound:

[0507]

[0508] Wherein the chiral center is in the D-configuration. This form of the compound may also be referred to as the DY stereoisomer of the compound of the present invention. The stereochemistry of the chiral center in the DY stereoisomer can be combined with other chiral centers of the compound (such as the chiral center in part A described herein) without restriction.

[0509] In any aspect of the invention, 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or more of the compound present in the composition is the R diastereomer around the chiral center indicated by * in portion A of the compounds described herein.

[0510] In any aspect of the invention, 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or more of the compound present in the composition is the S diastereomer around the chiral center indicated by * in portion A of the compounds described herein.

[0511] In any aspect of the invention, 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or more of the compound present in the composition is the R diastereomer around the chiral center indicated by ** in portion A of the compounds described herein.

[0512] In any aspect of the invention, 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or more of the compound present in the composition is the S diastereomer around the chiral center indicated by ** in portion A of the compounds described herein.

[0513] In any aspect of the invention, 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or more of the compound present in the composition is the R diastereomer around the chiral center indicated by *** in the Y moiety.

[0514] In any aspect of the invention, 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or more of the compound present in the composition is the S diastereomer around the chiral center indicated by *** of the Y moiety.

[0515] In any aspect of the invention, 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or more of the compound present in the composition is the R diastereomer around the chiral center indicated by * in portion A, the R diastereomer around the chiral center indicated by ** in portion A, and the R diastereomer around the chiral center indicated by *** in portion Y.

[0516] In any aspect of the invention, 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or more of the compound present in the composition is the R diastereomer around the chiral center indicated by * in portion A, the S diastereomer around the chiral center indicated by ** in portion A, and the R diastereomer around the chiral center indicated by *** in portion Y.

[0517] In any aspect of the invention, 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or more of the compound present in the composition is the R diastereomer around the chiral center indicated by * in portion A, the R diastereomer around the chiral center indicated by ** in portion A, and the S diastereomer around the chiral center indicated by *** in portion Y.

[0518] In any aspect of the invention, 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or more of the compound present in the composition is the R diastereomer around the chiral center indicated by * in portion A, the S diastereomer around the chiral center indicated by ** in portion A, and the S diastereomer around the chiral center indicated by *** in portion Y.

[0519] In any aspect of the invention, 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or more of the compound present in the composition is the S diastereomer around the chiral center indicated by * in portion A, the R diastereomer around the chiral center indicated by ** in portion A, and the R diastereomer around the chiral center indicated by *** in portion Y.

[0520] In any aspect of the invention, 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or more of the compound present in the composition is the S diastereomer around the chiral center indicated by * in portion A, the S diastereomer around the chiral center indicated by ** in portion A, and the R diastereomer around the chiral center indicated by *** in portion Y.

[0521] In any aspect of the invention, 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or more of the compound present in the composition is the S diastereomer around the chiral center indicated by * in portion A, the R diastereomer around the chiral center indicated by ** in portion A, and the S diastereomer around the chiral center indicated by *** in portion Y.

[0522] In any aspect of the invention, 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or more of the compound present in the composition is the S diastereomer around the chiral center indicated by * in portion A, the S diastereomer around the chiral center indicated by ** in portion A, and the S diastereomer around the chiral center indicated by *** in portion Y.

[0523] Compared to other related compounds, the compounds of the present invention exhibit improved solution stability under accelerated degradation conditions. Solution stability can be assessed by measuring the concentration of the compound in the solution at day 0 and comparing it with the concentration of the compound after a period of time (such as 14 days). Solution stability can be assessed under ambient conditions (such as 25°C and 65% relative humidity) or under accelerated conditions (such as 40°C and 75% relative humidity). Typically, the acceptable stability of a compound that is beneficial for the indications of the present invention is that the concentration of the compound in the solution will remain at least 80% after storage in the solution under accelerated conditions for 14 days relative to the initial concentration of the compound in the solution. Typically, the solution can be a saline solution (such as a 0.9% NaCl aqueous solution) or sulfate buffered saline (PBS; for example, pH 7.4). In some embodiments, the compound of the present invention is at least about 80%, 85%, 90%, 91%, 92% or more of the amount of the compound detected in the solution at day 0 after storage in a PBS buffer at pH 7.4 for 14 days.

[0524] "Prodrugs" are compounds that may not fully meet the structural requirements of the compounds provided herein, but are modified in vivo after administration to a subject or patient to produce a compound of the invention as described herein. For example, a prodrug may be an acylated derivative of a compound as provided herein. Prodrugs include compounds in which a hydroxyl, carboxyl, amine, or sulfhydryl group is bonded to any of the following groups, which, when administered to a mammalian subject, cleave to form free hydroxyl, carboxyl, amino, or sulfhydryl groups, respectively. Examples of prodrugs include, but are not limited to, acetate (salt), formate (salt), phosphate (salt), and benzoate (salt) derivatives of alcohol and amine functional groups in the compounds provided herein. Prodrugs of the compounds provided herein can be prepared in such a way that the functional groups present in the compound are modified so that the modifications are cleaved in vivo to produce the parent compound.

[0525] Prodrugs include compounds in which an amino acid residue, or a polypeptide chain of two or more (e.g., two, three, or four) amino acid residues is covalently linked to a free amino group and an amide group of a compound of the invention. Amino acid residues include 20 naturally occurring amino acids typically represented by three letter symbols, and also include 4-hydroxyproline, hydroxylysine, demosine, isodemosine, 3-methylhistidine, norvaline, β-alanine, γ-aminobutyric acid, citrulline, homocysteine, homoserine, ornithine, and methionine sulfone. Prodrugs also include compounds in which carbonates, carbamates, amides, and alkyl esters are covalently linked to the above-mentioned substituents of the compound of the invention.

[0526] The compounds of the present invention as described herein, or pharmaceutically acceptable salts, solvates or prodrugs thereof, may be covalent irreversible or covalent reversible agonists of the active site of a protein.

[0527] When referring to a protecting group (PG), one skilled in the art will readily understand what type of protecting group would be appropriate. Examples of suitable amine protecting groups for the purposes described herein include, but are not limited to, tert-butyloxycarbonyl (t-Boc) and 9H-fluoren-9-ylmethoxycarbonyl (Fmoc).

[0528] Pharmaceutical compositions can be formulated from the compounds of the present invention as described herein for any appropriate route of administration, including, for example, topical (e.g., transdermal or ocular), oral, buccal, respiratory (e.g., nasal, inhalation, intrapulmonary), vaginal, rectal, or parenteral administration. The term "parenteral" as used herein includes subcutaneous, intradermal, intravascular (e.g., intravenous), intramuscular, spinal, intracranial, intrathecal, intraocular, periocular, intraorbital, intrasynovial, and intraperitoneal injection, as well as any similar injection or infusion techniques. Suitable oral forms include, for example, tablets, troches, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules, or syrups or elixirs. For intravenous, intramuscular, subcutaneous, or intraperitoneal administration, one or more compounds can be mixed with a sterile aqueous solution that is preferably isotonic with the recipient's blood. Such preparations can be prepared in the following manner: the solid active ingredient is dissolved in water containing a physiologically compatible substance such as sodium chloride or sodium glycinate and having a buffered pH compatible with physiological conditions to prepare an aqueous solution, and then the solution is made sterile. These preparations can be present in unit dose containers or multidose containers (such as sealed ampoules or bottles). Examples of components are described in Martindale-The Extra Pharmacopoeia [Martindale's Pharmacopoeia] (Pharmaceutical Press, London, 1993) and Martin (editor), Remington's Pharmaceutical Sciences [Lemington's Pharmaceutical Sciences]. Preferably, these compositions are formulated for administration to the respiratory tract, for example, by intrapulmonary administration (e.g., inhalation) or intranasal administration. These compositions can be applied to the upper respiratory tract and / or lower respiratory tract.

[0529] Preferably, these pharmaceutical compositions are in a form suitable for administration via the respiratory route and may be in any form, such as a powder, liquid or suspension. Such compositions may target tissues including lung tissue (including alveoli, terminal bronchioles, bronchioles and bronchi) or nasal cavity (including paranasal cavity, frontal sinus, ethmoid sinus, maxillary sinus, sphenoid sinus, superior turbinate, middle turbinate and inferior turbinate).

[0530] In the context of this specification, the term "administering" and variations of the term include contacting a compound or composition of the invention with an organism or surface, or applying, delivering or providing the compound or composition of the invention to an organism or surface by any appropriate means.

[0531] The dosage of the biologically active compounds according to the invention can vary within wide limits and can be adjusted to the individual requirements.The active compounds according to the invention are generally administered in a therapeutically effective amount.

[0532] The compositions according to the present invention will be administered in an effective amount. The phrases 'therapeutically effective amount' or 'effective amount' generally refer to an amount of a compound of the present invention, a pharmaceutically acceptable salt, polymorph, or prodrug thereof, as described herein, that acts to: (i) treat a specific disease, condition, or disorder, (ii) alleviate, ameliorate, or eliminate one or more symptoms of a specific disease, condition, or disorder, or (iii) delay the onset of one or more symptoms of a specific disease, condition, or disorder as described herein. Sometimes, undesirable effects (e.g., side effects) may occur as well as the desired therapeutic effect; therefore, a practitioner will balance the potential benefits against the potential risks in determining what is an appropriate "effective amount."

[0533] The exact amount required will vary from subject to subject, depending on the species, age and general condition of the subject, the mode of administration, etc. Therefore, it may not be possible to specify an exact "effective amount". However, one of ordinary skill in the art can determine the appropriate "effective amount" in any individual case using only routine experiments. In one aspect, the dose administered to the subject is any dose that reduces the viral load. Preferably, the dose does not significantly increase inflammation, for example, does not significantly increase the absolute number of neutrophils in the lungs or the proportion of neutrophils to total BAL cells. The term "therapeutically effective amount" or "effective amount" may also refer to an amount of a compound of Formula (I), Formula (II), Formula (III), Formula (IV) and / or Formula (V) or a pharmaceutically acceptable salt, solvate or prodrug thereof that causes symptoms of respiratory infection, or improvement or repair of respiratory diseases or conditions associated with viral or bacterial infection.

[0534] In certain embodiments, the effective amount for a human subject is in the range of about 250nmol / kg body weight / agent to 0.005nmol / kg body weight / agent. Preferably, the range is about 250nmol / kg body weight / agent to 0.05nmol / kg body weight / agent. In certain embodiments, the body weight / agent range is about 250nmol / kg to 0.1nmol / kg, about 50nmol / kg to 0.1nmol / kg, about 5nmol / kg to 0.1nmol / kg, about 2.5nmol / kg to 0.25nmol / kg or about 0.5nmol / kg to 0.1nmol / kg body weight / agent. In certain embodiments, the amount is or is about a compound of 250nmol, 50nmol, 5nmol, 2.5nmol, 0.5nmol, 0.25nmol, 0.1nmol or 0.05nmol / kg body weight / agent. Dosage regimen is adjusted to adapt to emergency situations, and can be adjusted to obtain the optimal therapeutic dose.

[0535] The compounds of the present invention as described herein can be formulated as compositions for inhalation preparations (including dry powders, sprays, mists or aerosols). This may be particularly preferred for treating respiratory infections. For inhalation preparations, compositions or combinations provided herein can be delivered via any inhalation method known to those skilled in the art. Such inhalation methods and devices include, but are not limited to, metered dose inhalers with propellants such as CFC or HFA or physiologically and environmentally acceptable propellants. Other suitable devices are breath-operated inhalers, multi-dose dry powder inhalers and aerosol nebulizers. Aerosol formulations for the subject method typically comprise propellants, surfactants and cosolvents, and can be filled into conventional aerosol containers closed by suitable metering valves.

[0536] Inhalant compositions can comprise the liquid or powder composition containing active ingredient that is suitable for atomization and endobronchial use, or the aerosol composition used via the aerosol unit that distributes metered dose.Suitable liquid composition is included in the active ingredient in aqueous, pharmaceutically acceptable inhalation solvent such as isotonic saline or antibacterial water.By pump or extrusion-actuated atomizing spray dispenser or by any other conventional device application solution, to promote or make the liquid composition of required dose be sucked into patient's lung.Be suitable for, for example, as nasal spray or as nasal drops and the preparation (wherein carrier is liquid) that uses comprises aqueous or oily solution of active ingredient.Alternatively, composition can be dry powder and can be applied to respiratory tract as defined herein.

[0537] It will be understood that the specific dosage level for any particular patient will depend on a variety of factors, including the activity of the specific compound used, age, weight, general health, sex, diet, time of administration, route of administration, rate of excretion, drug combination (i.e., other drugs used to treat the patient), and the severity of the specific disorder being treated.

[0538] However, it should be understood that the specific dosage level for any particular subject will depend on a variety of factors, including the activity of the specific compound used, age, body weight, general health, sex, diet, administration time, administration route, excretion rate, drug combination (i.e., other drugs for treating the subject), and the severity of the specific disorder being treated. If the compound is topically applied rather than systemically, and the purpose of being applied is for prevention rather than treatment, the dosage will generally be lower. Such treatments can be applied frequently as needed and are applied for a time period judged to be necessary by the treating physician. Those skilled in the art will understand that for each individual, it may be necessary to optimize the dosage regimen or therapeutically effective amount of the compound of the present invention to be applied, or its pharmaceutically acceptable salt, solvate or prodrug. These pharmaceutical compositions may be included in the range of about 0.1 mg to 2000 mg, preferably in the range of about 0.5 mg to 500 mg, and most preferably in the range of about 1 mg to 200 mg of active ingredient. A daily dose of about 0.01 mg / kg body weight to 100 mg / kg body weight, preferably between about 0.1 mg / kg body weight and about 50 mg / kg body weight, may be suitable. The daily dosage can be administered as a single dose or in multiple doses per day.

[0539] It will also be understood that different dosages may be required to treat different disorders.

[0540] As used herein, the term "treating" or "treating" a subject includes applying or administering a compound or composition of the invention to a subject (or applying or administering a compound of the invention to cells or tissues of a subject) for the purpose of delaying, slowing, stabilizing, curing, healing, alleviating, relieving, altering, correcting, reducing worsening, alleviating, improving, or affecting a disease or condition, a symptom of a disease or condition, or the risk of (or susceptibility to) a disease or condition. The term "treating" refers to any sign of successful treatment or improvement of an injury, lesion, or condition, including any objective or subjective parameter, such as elimination; alleviation; slowing the rate of exacerbation; lessening the severity of a disease; stabilizing, alleviating symptoms, or making an injury, lesion, or condition more tolerable to the subject; slowing the rate of degeneration or decline; making a degenerative endpoint less debilitating; or improving the physical or mental health of the subject.

[0541] As used herein, "prevent" or "preventing" means at least reducing the likelihood of developing the risk (or susceptibility) of developing a disease or disorder (i.e., so that at least one of the clinical symptoms of the disease does not develop in a patient who may be exposed to or susceptible to the disease but has not yet developed or exhibited symptoms of the disease). Biological and physiological parameters for identifying such patients are provided herein and are also well known to physicians.

[0542] "Subject" includes any human or non-human animal. Thus, in addition to being useful for human therapy, the compounds of the present invention may also be used in veterinary therapy of mammals, including companion animals and farm animals, such as, but not limited to, dogs, cats, horses, cattle, sheep, and pigs.

[0543] The compounds of the present invention may be administered together with a pharmaceutical carrier, diluent or excipient as described above.

[0544]

[0545]

[0546]

[0547]

[0548]

[0549]

[0550]

[0551]

[0552]

[0553]

[0554] It should be understood that the invention disclosed and defined in this specification extends to all alternative combinations of two or more of the individual features mentioned or evident from the text and drawings. All these different combinations constitute multiple alternative aspects of the invention.

[0555] Example 1 - Synthesis of Compounds

[0556] Example 1.1 - Synthesis of A using Fmoc solid phase chemistry

[0557] Compounds of the present invention, including those according to formula (I), may be provided by coupling a compound of formula AI:

[0558]

[0559] Among them, L1, L2, Z1, Z2, v, b, w, z, R x 、R y 、R 11 、R 12 、R 13 、R 14 、R 15 、R 16 、R 17 、R 18 and X have the meanings as defined herein for any compound of the invention, and R 19 is an amino protecting group

[0560] and compounds having the formula YB-I:

[0561]

[0562] in

[0563] Y' is

[0564]

[0565] wherein R1 and R2 are independently selected from the group consisting of H, -CH2OH, -CH2CH2OH, -CH(CH3)OH, -CH2OPO(OH)2, -CH2C(=O)NH2, -CH2CH2C(=O)OH, and -CH2CH2C(=O)OR8, wherein any of the alkyl hydrogens may be substituted by halogen;

[0566] R8 is selected from the group consisting of H and linear or branched C1-C6 alkyl;

[0567] B' is polyethylene glycol (PEG); and

[0568] It is a solid carrier resin.

[0569] In some embodiments, B' comprises a substituted PEG having formula BI. In these embodiments, a solid phase reaction of the following sequence can be employed:

[0570] a) coupling 1 to 10 alpha amino acids or compounds derived from natural alpha amino acids (constituting L) to a solid phase resin, optionally using Fmoc chemistry

[0571] b) If L is present, PG-NH-(CH2) p -O-(CH2CH2O) n -(CH2) m-COOH is coupled to a solid phase resin or substituted resin, where PG represents an amino protecting group compatible with Fmoc chemistry;

[0572] c) removal of PG;

[0573] d) Coupling PG-NH-CR 13 R 14 -COOH, where PG' represents an amino protecting group compatible with Fmoc chemistry;

[0574] e) removal of PG';

[0575] f) coupling an acid having formula (AI);

[0576] g) optionally removing R 19 and optionally undergo acylation and / or alkylation to introduce R 18 and / or R 19 ;as well as

[0577] h) removing the compound from the solid support

[0578] In some embodiments, B' comprises a substituted PEG according to formula (B-II), and the following sequence of solid phase reactions can be employed:

[0579] a) coupling 1 to 10 alpha amino acids or compounds derived from natural alpha amino acids (constituting L) to a solid phase resin, optionally using Fmoc chemistry

[0580] b) If L is present, PG-NH-(CH2) t -O-(CH2CH2O) k -(CH2) h -COOH is coupled to a solid phase resin or substituted resin, where PG represents an amino protecting group compatible with Fmoc chemistry;

[0581] c) removal of PG;

[0582] d) Coupling PG'-NH-(CH2) p -O-(CH2CH2O) n -(CH2) m -COOH, where PG' represents an amino protecting group compatible with Fmoc chemistry;

[0583] e) removal of PG';

[0584] f) Coupling PG"-NH-CR 13 R 14 -COOH, where PG" represents an amino protecting group compatible with Fmoc chemistry;

[0585] g) Remove PG";

[0586] h) coupling an acid of formula (AI);

[0587] i) optionally removing R 19 and optionally acylated and / or alkylated to incorporate R 18 and / or R 19 ;as well as

[0588] j) removing the compound from the solid phase resin.

[0589] It will be understood that the exact sequence of events may vary from that outlined, and that additional steps may be added where necessary and synthetically advantageous, such as oxidation of cysteine ​​sulfur to sulfoxide.

[0590] Example 1.2 - Synthesis of Intermediates for Solid Phase Coupling A

[0591] Some examples of intermediate acids having formula A-II:

[0592]

[0593] Among them, L1, L2, X, v, w and R 18 is as defined for the compound of formula AI above, Z1 and Z2 are independently selected from -NHC(O)-, -C(O)NH-, -OC(O)-, -C(O)O-, -NHC(O)O- and -OC(O)O-;

[0594] It can be prepared by the synthesis shown in Scheme 1.

[0595] Scheme 1 describes the synthesis of examples of compounds having formula A-II, wherein

[0596] X is S,

[0597] L1-Z1 is -OC(O)EC g’ H (g’+2) , wherein E is -O- or -NH-, and g' is 11, 12, 13, 14, 15, 16, 17, 18 or 19;

[0598] L2-Z2 is -OC(O)EC g’ H (g’+2) , wherein E is -O- or -NH-, and g' is 11, 12, 13, 14, 15, 16, 17, 18 or 19; and

[0599] R 19 is PG3, which is an amino protecting group.

[0600] Solution 1

[0601]

[0602] The reaction of a protected enol of formula (V') wherein PG is a suitable protecting group (e.g., a silyl group such as TBDMS) forms an epoxide of formula (VI'). It will be appreciated that the formation of the epoxide can be carried out to give a racemic product or to give an enantiomerically enriched material. If a racemic or non-racemic mixture of enantiomers is produced, preparative chiral chromatography is employed to separate the enantiomers, if desired.

[0603] The epoxide of formula (VI') is reacted with an appropriately protected cystine analog, such as N-(((9H-fluoren-9-yl)methoxy)carbonyl)-S-(((R)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(tert-butyloxy)-3-oxopropyl)thio)-D-cysteine ​​tert-butyl ester, under reducing conditions to provide the alcohol of formula (VII') wherein PG2 is tert-butyl ester and PG3 is Fmoc. It will be appreciated that the alcohol of formula (VII') may consist of more than one stereoisomer, and in the event that multiple stereoisomers are present, they may be separated by chiral preparative chromatography as desired.

[0604] The alcohol of formula (VII') can be acylated using a suitable reagent to give a carbonyl-containing adduct of formula (VIII'). Where an ester is desired, the acyl chloride can be reacted in the presence of a suitable base and solvent; where a carbamate is desired, the isocyanate can be reacted in the presence of a suitable base and solvent, and where a carbonate is desired, the chloroformate can be reacted in the presence of a suitable base and solvent. The carbonyl-containing adduct of formula (VIII') can then be deprotected using a suitable reagent to expose the carboxylic acid of formula (IX'), for example, where PG2 is tert-butyl, trifluoroacetic acid can be used to preferably remove the tert-butyl group.

[0605] The acid of formula (IX') can then be used as a reagent in solid phase synthesis to add groups of formula Y and B.

[0606] Example 1.3 - Synthesis of B using Fmoc solid phase chemistry

[0607] Compounds of the present invention, including those according to formula (I) (wherein z is 1, w is 1, and b is 0), can be provided by preparing a resin-bound peptide having the formula:

[0608]

[0609] in

[0610] Y' is

[0611]

[0612] wherein R1 and R2 are independently selected from the group consisting of H, -CH2OH, -CH2CH2OH, -CH(CH3)OH, -CH2OPO(OH)2, -CH2C(=O)NH2, -CH2CH2C(=O)OH, and -CH2CH2C(=O)OR8, wherein any of the alkyl hydrogens may be substituted by halogen;

[0613] R8 is selected from the group consisting of H and linear or branched C1-C6 alkyl;

[0614] B' is polyethylene glycol (PEG);

[0615] PG s is H or a sulfur protecting group, such as tert-butyl; and

[0616] It is a solid carrier resin.

[0617] After optional sulfur deprotection, the resin-bound peptide can be reacted with a 1,2-epoxy-alkanol having the formula:

[0618]

[0619] where R x 、R y and v have the meanings given for formula (I),

[0620] to provide an alkylated thiol having formula S-1:

[0621]

[0622] S-1

[0623] wherein Y' and B' have the meanings given above, and v has the meaning given for the compound of formula (I), or its sulfone or sulfoxide.

[0624] The diol portion of the resin-bound compound S-1 can be further reacted (eg, by diol functionalization with a palmitic acid group or a lauryl carbamate group, etc.) to provide compounds of the present invention.

[0625] Example 1.4 - Synthesis and Characterization of Compounds 3, 4, 15 and 16

[0626] Synthesis of Compounds 3 and 4. The synthesis of Compounds 3 and 4 is described in Scheme 2 below.

[0627] Fmoc-Gly was added as the first amino acid to the solid support, followed by coupling with a 2-fold molar excess of Fmoc-NHCH2CH2O-(PEG) in the presence of a 2-fold excess of benzotriazole tetramethylurea hexafluorophosphate (HBTU), hydroxybenzotriazole (HOBT) and a 4-fold excess of diisopropylethylamine (DIPEA) in 2 ml of dimethylformamide (DMF). 11 -CH2CH2COOH or Fmoc-NHCH2CH2O-(PEG) 27 -CH2CH2COOH for 2 hours. Fmoc-Ser(tBu)-OH was then coupled to provide intermediate A2, followed by coupling of Boc-Cys(StBu)A1. The thiol tert-butyl group on the cysteine ​​residue was removed by incubating the peptide resin in 0.5 M dithiothreitol in DMF for 1 hour at room temperature. The saturated Boc-Cys-Ser(tBu)CH2CH2O-(PEG) in DMF was added to 250 mg of saturated Boc-Cys-Ser(tBu)CH2CH2O-(PEG) 11 -CH2CH2C(O)Gly resin or Boc-Cys-Ser(tBu)CH2CH2O-(PEG) 27 250 μl of R-(+)-1,2-epoxy-butane-4-ol [(R)-2-(oxiran-2-yl)ethane-1-ol] (M-CH2CH2C(O)Gly resin (0.25 mmole / g, 0.25 g = 0.0625 mmole) was added to W = 88.11, d = 1.1, 250 μl = 3.125 mmol equivalent to a 50-fold excess of free thiol groups present on the peptide resin) and 25 μl of diisopropylethylamine (DIPEA, M W = 129.2, d = 0.74, 25 μl = 0.14 mmol). The reaction mixture was placed in a 50°C water bath for 2 hours, and then washed thoroughly with DMF to provide intermediate A3.

[0628] Palmitic acid (320 mg, 1.25 mmol), DIPCDI (225 uL, 1.5 mmol) and 4-dimethylaminopyridine (DMAP; 15.25 mg, 0.125 mmol) were dissolved in 2 mL of dichloromethane (DCM) at room temperature and added to resin-bound BOC-Dhc-peptide resin A3 (0.0625 mmol, 0.25 g) and shaken for 16 h. The supernatant was removed by filtration and the solid support was thoroughly washed with DCM and dimethylformamide (DMF) to remove any residues of urea before being subjected to the cleavage process described below.

[0629] The solid support with the assembled lipopeptide was exposed to Reagent B (93% TFA, 5% water, and 2% triisopropylsilane) for 2 hours. To isolate the product, most of the TFA was removed and the residue was then dissolved in 50% acetonitrile and immediately purified using the purification protocol described below, or the material was freeze-dried and stored for later purification.

[0630] Scheme 2. Synthesis of Compound 3 (x = 11) and Compound 4 (x = 27)

[0631]

[0632] Synthesis of Compound 15 and Compound 16. The synthesis of Compounds 15 and 16 was carried out as described in Scheme 3. Intermediate A3 was prepared as described for Compounds 3 and 4 above.

[0633] Then, 100 μl of ethyl methyl sulfide (M W =76.16, d=0.842, 100 μl=1.10 mmol), followed by 105 μl of tetradecyl isocyanate (MW=239, d=0.869, 105 μl=0.38 mmol, i.e., a 3-fold excess per hydroxyl group present on the solid support), and finally 210 μl of dibutyltin dilaurate (M W =631.6, d=1.053, 210 μl=0.35 mmol). The reaction mixture was purged with nitrogen for approximately 5 min and mixed (Intelli-Mixer, RM-2, using program F26) overnight at room temperature. The reaction mixture was transferred to a 50 ml tube and chloroform was added to 50 ml. After approximately 5 min of sonication, the white precipitate formed during the reaction dissolved. The solid support was washed with DMF and acetonitrile and the final product obtained after cleavage of the support (as above) was purified by HPLC.

[0634] Scheme 3. Synthesis of compounds 15 (x = 11) and 16 (x = 27)

[0635]

[0636] Synthesis of Compound 20. Compound 20 was synthesized by standard Fmoc solid phase peptide synthesis starting from Fmoc-RINK MBHAPS resin. Removal of the Fmoc group was achieved using 20% ​​piperidine in DMF after each coupling. Fmoc-Gly-OH (2-fold excess), Fmoc-NH-PEG-H-yl-D-glycine (2-fold excess), and Fmoc-NH-PEG-H-yl-D-glycine (2-fold excess) were coupled in DMF using an equivalent excess of ethyl cyano(oxime)acetate (Oxyma Pure) and diisopropylcarbodiimide (DIC) as coupling agents. 28-CH2CH2COOH (1.4-fold excess), Fmoc-Ser(tBu)-OH (2-fold excess) and N-(Boc)-S-((R)-2,3-dihydroxybutyl)-L-cysteine ​​(1.5-fold excess). At room temperature, the coupling of tetradecyl chloroformate (12 equivalents compared to moles of resin) and DIEA (24 equivalents compared to moles of resin) in anhydrous DCM was carried out for 18 hours. This coupling was repeated three times (recoupling). At room temperature, the first coupling was carried out using tetradecyl chloroformate (12 equivalents compared to moles of resin) and NMM (24 equivalents compared to moles of resin) in anhydrous DCM / THF (85 / 15) for 18 hours. The second coupling was performed at room temperature using tetradecyl chloroformate (6 equivalents compared to the mole resin), NMM (12 equivalents compared to the mole resin) in anhydrous DCM / THF (85 / 15) for 41 hours. Finally, the third coupling was performed at room temperature using tetradecyl chloroformate (6 equivalents compared to the mole resin), NMM (12 equivalents compared to the mole resin) in anhydrous DCM / THF / toluene (85 / 15 / 5) for 21.5 hours.

[0637] The peptide was cleaved from the resin, the N-terminal Boc group was removed, and the serine side chain was deprotected by exposing the resin to a solution of 93% trifluoroacetic acid (TFA), 5% H2O, 3% triisopropylsilane (TIPS) for 1.5 hours. After the cleavage reaction, the mixture was evaporated and the resulting residue was redissolved in 30% acetonitrile / water and lyophilized.

[0638] Synthesis of compound 24. Compound 24 was synthesized by standard Fmoc solid phase peptide synthesis starting from chlorotrityl chloride resin with an initial substitution of 1.6 meq / g. The first amino acid Fmoc-Gly-OH was first loaded onto the resin using a 0.5-fold molar excess of Fmoc-Gly-OH and DIEA (1.5-fold excess), followed by capping with DMF / MeOH / DIEA (80 / 10 / 10) and Fmoc deprotection to obtain dry loaded H-Gly-CT resin with a final substitution of 0.67 meq / g. Removal of the Fmoc group was achieved after each coupling using 20% ​​piperidine in DMF. Fmoc-NH-PEG was reacted with (7-azabenzotriazol-1-yloxy)tripyrrolidinephosphonium hexafluorophosphate (PyAOp; 1.4 equiv), diisopropylethylamine (DIEA; 3.2 equiv) in DMF. 28The coupling of Fmoc-Ser(tBu)-OH (2 equiv) and N-(Boc)-S-((R)-2,4-dihydroxybutyl)-L-cysteine ​​(1.5 equiv) was performed in DMF using a considerable excess of Oxyma Pure and DIC as coupling agents. Palmitic acid coupling was performed using palmitic acid (20 equiv, compared to the molar resin), DIC (20 equiv), DMAP (2 equiv) in DCM / THF (85 / 15) (v / v) at room temperature for 24 hours.

[0639] The peptide was cleaved from the resin, the N-terminal Boc group was removed, and the serine side chain was deprotected by exposing the resin to a solution of 93% TFA, 5% H2O, 3% TIPS for 1.5 hours. After the cleavage reaction, the mixture was evaporated and the resulting residue was redissolved in 30% acetonitrile / water and lyophilized.

[0640] Synthesis of compound 36. Compound 36 was synthesized by standard Fmoc solid phase peptide synthesis starting from Fmoc-RINK MBHA PS resin. Removal of the Fmoc group was achieved using 20% ​​piperidine in DMF after each coupling. Fmoc-Gly-OH (2-fold excess), Fmoc-NH-PEG were coupled in DMF using an equivalent excess of Oxyma Pure and DIC as coupling agents. 28 -CH2CH2COOH (1.4-fold excess), Fmoc-Ser(tBu)-OH (2-fold excess) and N-(Boc)-S-((R)-2,3-dihydroxybutyl)-L-cysteine ​​(1.5-fold excess). 2-Methyl-palmitic acid coupling was performed at room temperature in DCM / THF (85 / 15) (v / v) using 2-methyl-palmitic acid (20 equiv. compared to the molar resin), DIC (20 equiv.), DMAP (2 equiv.) for approximately 20 hours.

[0641] The peptide was cleaved from the resin, the N-terminal Boc group was removed, and the serine side chain was deprotected by exposing the resin to a solution of 93% TFA, 5% H2O, 3% TIPS for 1.5 hours. After the cleavage reaction, the mixture was evaporated and the resulting residue was redissolved in 30% acetonitrile / water and lyophilized.

[0642] Purification and characterization:

[0643] Purification and characterization: After cleavage from the solid support, each analog was purified by reverse phase HPLC according to Protocol A or Protocol B as described below.

[0644] Protocol A: Reversed-phase HPLC was performed using an Agilent Zorbax 300SB-C3, 5 μm column (9.4 mm × 250 mm; Agilent Technology, Australia) installed in an Agilent HPLC 1260 Infinity system (Agilent Technologies, Santa Clara, CA, USA), and chromatograms were generated using buffer A (0.1% trifluoroacetic acid in water) and buffer B (0.1% trifluoroacetic acid in acetonitrile).

[0645] Protocol B: Reverse phase chromatography was performed using a Novasep axial compression column (5 cm diameter) loaded with cyano media (Daisogel SP-120-CN-P) with an acetonitrile gradient in [0.1% TFA / water]. After lyophilization of the intermediate, ion exchange was performed on Dowex ion exchange resin to obtain the peptide as an acetate salt.

[0646] Compounds 3, 4, 15, and 16 were prepared as described above and purified by Protocol A, and compounds 20, 24, and 36 were prepared as described above and purified by Protocol B.

[0647] The identity and purity of the target material were determined using an inline HPLC-MS system using the following conditions:

[0648] Conditions A: HPLC column: Agilent Zorbax 300-SB C3 (150×0.5 mm; 5 μm) with the following gradient conditions: 0-5 min, 20% B: 5-32 min, 20% B-100% B: 32-40 min, 100% B-20% B. Flow rate: 20 μl / min. LC-MS: Agilent 1100 Series capillary LC system inline with an Agilent 1100 Series LC / MSD ion trap mass spectrometer. The mass spectrometer was operated with electrospray ionization set in positive ion mode. Data analysis software from Agilent Technologies was used to deconvolute the charged ion series for identification of the peptide material, and the material was then characterized by LC-MS.

[0649] Condition B: Analytical reversed-phase HPLC with a cyano column (Daiso Fine Chem, SP-120-3-CN-P, 150×4.6 mm, 3 μm, ). Peptides were also analyzed by ESI LC-MS in positive ion mode using a Finnigan LCQ Deca XPMax.

[0650] According to Condition A, compounds 3, 4, 15 and 16 prepared and purified as described above were each found to have a purity greater than 95%.

[0651] According to Condition B, compounds 20, 24 and 36 prepared and purified as described above were found to have a purity greater than 95%.

[0652] The experimental masses (m / z) were consistent with the calculated molecular weight of each compound.

[0653] Peptide quantification

[0654] Compounds 3, 4, 15, and 16 were quantified by vacuum hydrolysis of the samples at 110° C. in the presence of 6NHCI containing 0.1% phenol in sealed glass vials. Amino acid derivatization was then performed using Waters AccQTag reagent according to the manufacturer's instructions, followed by analysis using an AccQTag ultra column (2.1 mm × 100 mm; Millipore Waters) on a Waters Acquity UPLC system (Millipore Waters). Quantification of other compounds was achieved using a similar protocol.

[0655] 1.5-Synthesis of sulfone and sulfoxide analogs of compounds 15 and 16

[0656] Sulfone and sulfoxide derivatives of compounds 15 and 16 can be obtained by synthetic routes similar to those described above, except that the ethyl methyl sulfide scavenger and, optionally, the nitrogen purge are omitted from the carbamate formation step. This reaction can yield a mixture of thiol, sulfone, and sulfoxide derivatives, which can be separated and purified by HPLC.

[0657] Alternatively, sulfone or sulfoxide derivatives can be prepared by oxidation of the corresponding sulfide with an oxidizing agent such as m-chloroperbenzoic acid (MCPBA) or tert-butyl hydroperoxide (t-BuOOH) under appropriate conditions.

[0658] Example 2 - Activation of human TLR2

[0659] The compounds were tested in an in vitro assay for their efficacy as human and mouse TLR-2 activators. This assay assesses NF-κB activation in the HEKBlue-mTLR-2 cell line. These cells have been stably transfected with mouse TLR-2 and endogenously express TLR-1 and TLR-6 at levels sufficient to allow full functional activation of TLR-1 / 2 and TLR-2 / 6.

[0660] Toll-like receptor 2 (TLR2) stimulation was tested by assessing NF-κB activation in the HEKBlue-hTLR2 cell line. These cells have been stably transfected with human TLR2 and endogenously express TLR1 and TLR6 at levels sufficient to allow full functional activation of TLR1 / 2 and TLR2 / 6. The test article was tested for its activity as a potential agonist against human TLR2. The test article was evaluated at seven concentrations and compared to a control ligand. These steps were performed in triplicate.

[0661] NF-kB reporter gene assay protocol: This assay was performed as previously described (Jackson et al., 2004; Lau et al., 2006; Sandor et al., 2003; Zeng et al., 2010). HEK293T cells were plated at 4 × 10 4 Cells were cultured at a density of 100 cells / well in 96-well plates and 24 h later, transfected with 100 ng of a NF-κB luciferase reporter gene [50 ng of a TK-renilla-luciferase expression plasmid (Promega Corporation, Madison, WI, USA) with or without 5 ng of a TLR2 expression plasmid in the presence of 0.8 μl of Fugene 6 (Roche Diagnostic). Compounds were added to the wells 24 h later at the concentrations indicated in the histogram. 5 h after stimulation, cell lysates were prepared using a reporter lysis buffer (Promega Corporation, Madison, WI, USA). The luciferase activity was measured using a kit (Promega Corporation, Madison, WI, USA) and a FLUOstar microplate reader (BMG Labtech, Ottenberg, Germany). Luciferase activity in cell lysates was determined using Labtech. NF-κB-dependent firefly luciferase activity was normalized to NF-κB-independent Renilla luciferase activity. Relative stimulation was calculated as the ratio of stimulated to unstimulated samples.

[0662] The results of this test for compounds 3, 4, 15 and 16 were Figure 1 These data show that these compounds exhibit significant activity against TLR2.

[0663] Example 3 - URT virus challenge

[0664] In these examples, an upper respiratory tract (URT) influenza virus challenge model was used in mice, using a dose of infectious virus that replicates in the URT and then progresses to the lungs. The URT model was used to determine which compounds could block influenza virus replication and spread from the URT to the lungs.

[0665] Cytokine and chemokine profiles were also measured in the nasal turbinates, trachea, lungs, and serum of animals treated with three or a single dose of compound for URT.

[0666] The cytokine profiles of mice pretreated with three doses of a compound of the invention and then challenged with Udorn virus were also measured.

[0667] experimental animals

[0668] All studies were performed using groups of male or female C57BL / 6 mice of similar age (eg, approximately 6 to 8 weeks of age). Mice were monitored daily for changes in body weight and behavioral or physical changes following administration of saline, compound, or viral challenge.

[0669] URT administration of compounds

[0670] Mice were anesthetized by inhalation of isoflurane and saline or various doses of compound diluted in saline were administered intranasally using a pipette.For multiple treatment experiments, mice received 3 doses of the compound of the invention every other day over a 5-day period.

[0671] Preparation of influenza virus

[0672] Influenza A / Udorn / 307 / 72 (H3N2) virus (i.e., Udorn virus) was propagated in the allantoic cavity of 10-day-old embryonated chicken eggs. 3 The eggs were inoculated with pfu of virus. After incubation at 35°C for 2 days, the eggs were frozen at 4°C and the allantoic fluid was harvested and clarified by centrifugation. The viral infectivity titer (pfu / mL) was determined by plaque assay as described below, and aliquots of the allantoic fluid were stored at -80°C until needed.

[0673] URT virus challenge

[0674] Mice were anesthetized with isoflurane and inoculated intranasally with 500 pfu of Udorn virus in 10 μl of saline using a pipette.On day 5 after challenge, nasal turbinates, trachea, and lungs were harvested to assess viral load.

[0675] Extraction and preparation of homogenates of nasal conchae, trachea, and lungs

[0676] 24 hours after treatment or after influenza challenge 5 days by CO2 asphyxiation mice were put to death.The nasal concha, trachea and lung of each mouse were collected in 1.5mL RPMI-1640 culture medium containing antibiotics (100ug / mL penicillin, 180ug / mL streptomycin and 24ug / mL gentamicin) and kept on ice until treatment.Tissue was homogenized using a tissue homogenizer, and the organ homogenate of the resulting was then centrifuged at 2,000rpm for 5 minutes to remove cell debris.Supernatant was collected and stored at -80°C for subsequent measurement.

[0677] Assessment of viral titer

[0678] The titer of infectious Udorn virus was determined by plaque assay on confluent monolayers of Madin Darby canine kidney (MDCK) cells. Six-well tissue culture plates were seeded with 3 ml of RP10 (RPMI-1640 medium supplemented with 10% (vol / vol) heat-inactivated FCS, 260 μg / mL glutamine, 200 μg / mL sodium pyruvate, and antibiotics) to a seeding density of 1.2 × 10 6 MDCK cells / well. At 37 ℃ in 5% CO 2 , after overnight incubation, the fused monolayer was washed with RPMI. The test supernatant serially diluted in RPMI containing antibiotics was added to the holes of the monolayer in duplicate. After incubation at 37 ℃ for 45 minutes in 5% CO 2 , the monolayer was covered with a covering culture medium containing 0.9% agarose and 2ug / mL trypsin-TPCK of 3mL, which was treated with glutamine and antibiotics in Leibovitz L15pH 6.8 culture medium. At 37 ℃ in 5% CO 2 , the culture plate was incubated for 3 days, and then the virus-mediated cell lysis was counted as the plaques on the cell layer. The total organ virus titer (plaque forming unit, PFU) of each animal was calculated.

[0679] Measurement of cytokine levels in nasal turbinates, trachea, lungs, and serum

[0680] IFN-γ, IL-2, IL-4, TNF, IL-10, IL-6, KC, MCP-1, RANTES, IL-12 / IL-23p40, and IL-17A were measured in nasal turbinates, trachea, lung homogenates, and serum samples using the BD Cytometric Bead Array (CBA) Flex kit according to the manufacturer's instructions, except that a total of 0.15 μl of each capture bead suspension and 0.15 μl of each PE-detection reagent were used per 50 μl sample. Samples were analyzed using a Becton Dickinson FACSCanto II flow cytometer, and data were analyzed using FCAP array multiplexing software.

[0681] Statistical analysis

[0682] One-way analysis of variance (ANOVA) with Tukey's test can be used, wherein all columns within the test are compared. Two-way analysis of variance (ANOVA) with Bonferroni's test can be used to compare the same treatment groups in the single dose regimen and the three repeated dose regimens. A p-value of ≤0.0322 is considered statistically significant. Statistical analysis is performed using appropriate software (e.g., GraphPad Prism, Version 7.0).

[0683] Example 4 - Evaluation of the Effect of Pretreatment with Various Doses of Compounds of the Invention on the Outcome of URT Challenge with Udorn Virus

[0684] This experiment was conducted to determine the antiviral effect of URT pretreatment with various doses of compounds of the invention.

[0685] On day 0, mice (5 animals / group) received saline, 5 nmol, 0.1 nmol, or 0.005 nmol of the compound of the invention in 10 μl intranasal administration under isoflurane anesthesia. On day 1 after administration of the compound of the invention, mice were challenged intranasally with 500 pfu of Udorn virus in a volume of 10 μl under isoflurane anesthesia. Mice were sacrificed on day 5, and the nasal turbinates, trachea, and lungs were removed, homogenized, and frozen for subsequent analysis.

[0686] The experimental design is summarized in the following schematic diagram

[0687]

[0688] Example 5 - Activation of TLR2 by Various Compounds

[0689] The ability of various compounds to stimulate luciferase activity in a NF-κB cell-based reporter system was compared. HEK293T cells transiently co-transfected with a human TLR2 plasmid and a luciferase-NF-κB plasmid reporter system were exposed to various dilutions of each compound. Successful receptor binding and subsequent signaling events were determined by measuring luminescence caused by luciferase activity.

[0690] Example 6 - TLR Binding and Specificity

[0691] The compounds of the present invention were evaluated for their ability to activate a range of other TLR pattern recognition receptors. These evaluations were performed using both human and mouse TLR panels. These assays detect a secreted embryonic alkaline phosphatase (SEAP) reporter gene controlled by a promoter that can be induced by NF-κB activation in HEK293 cells.

[0692] The secreted embryonic alkaline phosphatase (SEAP) reporter gene is controlled by a promoter that can be induced by the transcription factor NF-κB. This reporter gene allows for monitoring signal transduction through TLRs based on NF-κB activation. In a 96-well plate (200 μL total volume) containing appropriate cells (50,000-75,000 cells / well), 20 μL of test article or positive control ligand is added to the wells. The culture medium added to the wells is designed to detect NF-κB-induced SEAP expression. After incubation for 16-24 hours, the optical density (OD) is read at 650 nm on a Molecular Devices SpectraMax 340PC absorbance detector.

[0693] Control ligand

[0694] hTLR2: 1×108 cells / mL of HKLM (heat-inactivated Listeria monocytogenes)

[0695] hTLR3: 1μg / mL Poly(I:C)HMW

[0696] hTLR4: 100 ng / mL E. coli K12 LPS

[0697] hTLR5: 100 ng / mL of Salmonella typhimurium flagellin

[0698] hTLR7: 1 μg / mL CL307

[0699] hTLR8: 1 μg / mL CL075

[0700] hTLR9: 1 μg / mL of CpG ODN2006.

[0701] Example 7-Stability I

[0702] Stability was assessed by tracking the change in absolute peak area and % peak area of ​​the compounds of the invention relative to the peak area and % peak area obtained from freshly prepared solutions of the related compounds subjected to the following conditions. The compounds were formulated into each of the following formulations:

[0703] 1. Phosphate buffered saline (PBS), pH 7.4. For example, PBS buffer may contain 8 g NaCl, 0.2 g KCl, 1.15 g disodium hydrogen phosphate, and 0.2 g potassium dihydrogen phosphate in 1 liter of MilliQ water.

[0704] 2. 0.9% w / w saline (pH 5.8). For example, saline solution can be prepared by dissolving sodium chloride (1.855 g) in 200 mL of Milli-Q water.

[0705] The stability of each formulation was evaluated under the following conditions:

[0706] 1. 25℃ / 60% relative humidity (ICH environment)

[0707] 2. 40°C / 75% relative humidity (ICH accelerated)

[0708] Sample preparation

[0709] A solution of approximately 1 mg / mL of each compound (2 mL) was prepared in a PBS and saline diluent system. The actual concentrations are provided in Table 1.

[0710] Table 1: Test solution concentrations used for stability monitoring

[0711] Compound number diluent Concentration (mg / mL) 3 brine 1.05 PBS 1.10 4 brine 1.10 PBS 1.05 15 brine 1.05 PBS 1.03 16 brine 0.990 PBS 0.956

[0712] All compounds were heated to approximately 60°C under running hot tap water for approximately 30 seconds, followed by vortex mixing for an additional 30 seconds.

[0713] Each vial in Table 1 was further divided into 3 separate HPLC vials, which were then stored at 4°C to 8°C (refrigerator), 25°C / 65%RH, and 40°C / 75%RH for 2 weeks. The vials were wrapped in aluminum foil to protect from light during storage.

[0714] Equipment and operating parameters

[0715] A Shimadzu Nexera UHPLC with a diode array detector was used to monitor the peak area changes at t = 0 and t = 2 weeks.

[0716] Impurity and degradant peaks were identified using a Shimadzu LCMS-8030 system, and the selectivity of the HPLC method was determined by examining the main HPLC peaks for all possible coeluting components.

[0717] UHPLC parameters

[0718] Column - Phenomenex Kinetex Biphenyl, 50 x 2.1 mm, 2.6 μm, part number 00B-4622-AN

[0719] Vials - Agilent clear glass, 2 mL, with multiple injection septa, part number 226-50512-00

[0720] Mobile phase A - 5 mM ammonium formate in Milli-Q water

[0721] Mobile phase B - acetonitrile, Merck LC-MS grade

[0722]

[0723] Table 2: Gradient 1

[0724] Time (min) ％A ％B Start 55 45 0.1 55 45 8.0 25 75 8.5 25 75 8.6 55 45

[0725] Table 3: Gradient 2

[0726] Time (min) ％A ％B Start 55 45 0.1 55 45 8.0 25 65 8.5 25 65 8.6 55 45

[0727] LCMS parameters

[0728]

[0729]

[0730] Stability results

[0731] Samples of each of Compounds 3, 4, 15, and 16 were analyzed using gradient 1 (Compounds 3 and 4) or 2 (Compounds 15 and 16). The results are shown in Tables 4 and 5 below and Figure 2 and 3 The storage results at 40°C are shown in Table 4, and the storage results at 25°C are shown in Table 5.

[0732] Stability results compare the peak areas of samples subjected to the relevant conditions to the percent peak area of ​​the main HPLC peak at time zero, where the latter is normalized to 100% to exclude the contribution of impurities to the total peak area. Figure 2 Data for week 2 calculated by this method are shown. Figure 3A second data set generated using the main peak area, expressed as a percentage of the area at time zero, is shown.

[0733] Table 4: Stability of compounds 3, 4, 15 and 16 in saline (0.9%) or PBS (pH 7.4) at 40°C over time

[0734]

[0735] Table 5: Stability of compounds 3, 4, 15 and 16 in saline (0.9%) or PBS (pH 7.4) at 25°C over time

[0736]

[0737]

[0738] Compounds 3, 4, 15, and 16 each exhibited a great degree of stability over the 2-week testing period, indicating that they may be reasonably stable upon storage and administration.

[0739] Example 8-Stability II

[0740] The relative stability of compounds 4, 16, 20, 24, and 36 and the relative stability of compound (8) in WO2019 / 119067 were evaluated under accelerated conditions (40°C / 75% RH) for 9 days. Each compound was prepared at a concentration of 1 mg / mL in an aqueous formulation of 0.1% w / v EDTA / 0.9% w / v saline buffered to pH 5.

[0741] The structure of compound (8) in WO2019 / 119067 is:

[0742]

[0743] Stability was measured using reversed-phase HPLC with a UV detector (analytical wavelength of 205 nm). The peak area of ​​each compound at the 9th day time point was compared to the area at time zero. Compound stability was calculated as the percentage of the peak area data at time zero at day 9.

[0744] Compound stability was further assessed by comparison with a reference sample of the same compound. The reference sample was prepared at a concentration of 1 mg / mL in an aqueous formulation of 0.1% w / v EDTA / 0.9% w / v saline buffered to pH 5 and frozen for the duration of the test. Thawed samples were sonicated and measured by HPLC.

[0745] The data for each compound are summarized in Table 6.

[0746] Table 6

[0747]

[0748]

[0749] Compounds 4, 16, 20, 24, and 36 each showed a great degree of stability over the 9-day testing period. Each of these compounds also showed better stability under accelerated conditions than the comparative compound.

[0750] The stability of the compound over further extended periods of time is assessed by extending the exposure to accelerated conditions (eg, for 28 days).

[0751] Example 9 - Activation of human TLR2II

[0752] The compounds were tested for their potency as human TLR-2 activators in an in vitro assay in HEK-BLUE-hTLR2 cells.

[0753] Culture of HEK-BLUE-hTLR2 cells

[0754] HEK-BLUE-hTLR2 cells are designed to study human TLR2 (hTLR2) stimulation by monitoring NF-κB activation. HEK-BLUE-hTLR2 cells are generated by co-transfecting hTLR2 and a SEAP (secreted embryonic alkaline phosphatase) reporter gene into HEK293 cells. Stimulation with TLR2 ligands activates NF-κB, which in turn leads to SEAP production.

[0755] HEK-BLUE-hTLR2 cells were purchased from InvivoGen, San Diego, California, USA. Cells were grown in DMEM supplemented with 10% FCS, 100 U / ml penicillin, 100 μg / ml streptomycin, 2 mM L-glutamine, and 100 μg / mL Normocin in the presence of selective antibiotics purchased from InvivoGen and passaged at 70% confluency according to the manufacturer's recommendations. Cells were removed and resuspended in assay medium for testing according to the manufacturer's recommendations.

[0756] Testing of compounds

[0757] i) Prepare serial dilutions of the respective compound in saline and add 20 ml of each dilution in triplicate to each well of a flat-bottom 96-well plate and place it in the incubator while waiting for the cells.

[0758] ii) Remove the HEK-BLUE-hTLR2 cells in the T-75 flask from the incubator and discard the growth medium.

[0759] iii) Gently rinse the cells with 10 ml of pre-warmed PBS.

[0760] iv) Add 5 ml of pre-warmed PBS and place the cells at 37°C for 2 min, and then detach the cells by gently dripping PBS up and down on the surface where the cells are adhered.

[0761] v) In HEK-Blue TM A cell suspension was prepared at a density of 280,000 cells / ml in assay medium purchased from InvivoGen and prepared according to the manufacturer's instructions.

[0762] vi) Immediately add 180 ml of the cell suspension to each well of the plate containing the compound solution. The plate is then returned to the 37°C incubator for 16 hours and read at 620 nm using an ELISA reader.

[0763] The results of this test for compounds 4, 20, 24 and 36 were Figure 4 These data show that these compounds exhibit significant activity against TLR2.

Claims

1. A compound of formula (VI) or a pharmaceutically acceptable salt thereof: in n is 3 to 100; m is 1, 2, 3 or 4; p is 2, 3, or 4; q is zero or 1; R1 and R2 are independently selected from the group consisting of H, -CH2OH, -CH2CH2OH, -CH(CH3)OH, -CH2OPO(OH)2, -CH2C(=O)NH2, -CH2CH2C(=O)OH, and -CH2CH2C(=O)OR8, wherein any of the alkyl hydrogens may be substituted by halogen; R8 is selected from the group consisting of H and linear or branched C1-C6 alkyl; wherein when q=1, R3 is -NH2 or -OH; Wherein when q=0, R3 is H; L is zero or consists of 1 unit, where each unit has the formula: wherein R4 is H; and R5 is H, b is 0; w is 1; v is 2 or 3; z is 1; X is -S-; Z1 and Z2 are each independently selected from the group consisting of: -O-, -NR-, -S-, -S(=O)-, -S(=O)2-, -C(=O)O-, -OC(=O)-, -C(=O)NR-, -NRC(=O)-, -C(=O)S-, -SC(=O)-, -OC(=O)O-, -NRC(=O)O-, -OC(=O)NR-, and -NRC(=O)NR-; In each case of b, v, w, and z, R 11 、R 12 、R x 、R y 、R 14 、R 15 、R 16 and R 17 Each independently is H or a C1-C6 aliphatic group; R, R 13 and R 18 Each independently is H or a C1-C6 aliphatic group; R 19 is H or a C1-C6 aliphatic group; L1 and L2 are each independently C5-C 21 Aliphatic or C4-C 20 heteroaliphatic groups; or a pharmaceutically acceptable salt thereof.

2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein q is 1.

3. The compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, wherein n is from 10 to 14.

4. The compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, wherein n is 11.

5. The compound of claim 1 or 2, or a pharmaceutically acceptable salt thereof, wherein n is from 24 to 30.

6. The compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, wherein n is 27.

7. The compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, wherein m is 1, 2 or 3.

8. The compound according to claim 6, or a pharmaceutically acceptable salt thereof, wherein m is 2.

9. The compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, wherein R x 、R y 、R 11 、R 12 、R 13 、R 14 、R 15 、R 16 and R 17 It’s H.

10. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein Z1 and Z2 are the same and are selected from the group consisting of: -O-, -NR-, -S-, S(=O), S(=O)2-, -C(=O)O-, -OC(=O)-, -C(=O)NR-, -NRC(=O)-, -C(=O)S-, -SC(=O)-, OC(=O)O-, NRC(=O)O-, -OC(=O)NR-, and -NRC(=O)NR-, wherein each R is independently H or a C1-C6 aliphatic group.

11. The compound of claim 1 or 2, or a pharmaceutically acceptable salt thereof, wherein the compound is the R stereoisomer of the compound around the chiral center indicated by *: and / or The chiral center represented by ** in the following molecular formula is of L configuration: and / or The chiral center represented by *** in the following molecular formula is of L configuration:

12. A compound selected from any one of the following compounds or a pharmaceutically acceptable salt thereof.

13. A compound selected from any one of the following compounds: or a pharmaceutically acceptable salt thereof.

14. A composition comprising the compound according to any one of claims 1 to 13 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, diluent or excipient.

15. Use of a compound according to any one of claims 1 to 13, or a pharmaceutically acceptable salt thereof, in the preparation of a medicament for enhancing an innate immune response in a subject.

16. Use of the compound according to any one of claims 1 to 13, or a pharmaceutically acceptable salt thereof, in the preparation of a medicament for treating and / or preventing a disease caused by an infectious agent.

17. Use of a compound according to any one of claims 1 to 13, or a pharmaceutically acceptable salt thereof, in the preparation of a medicament for treating and / or preventing a respiratory disease or condition associated with a viral or bacterial infection in a subject.

18. Use of a compound according to any one of claims 1 to 13, or a pharmaceutically acceptable salt thereof, in the preparation of a medicament for treating and / or preventing a respiratory tract infection in a subject.

19. Use of the compound according to any one of claims 1 to 13, or a pharmaceutically acceptable salt thereof, in the preparation of a medicament for alleviating airway inflammation.

20. Use of a compound according to any one of claims 1 to 13, or a pharmaceutically acceptable salt thereof, in the preparation of a medicament for improving a subject's ability to control a respiratory disease or condition during a respiratory viral infection.

21. The use of claim 20, wherein the infection is not a rhinovirus infection.

22. Use of the compound according to any one of claims 1 to 13, or a pharmaceutically acceptable salt thereof, in the preparation of a medicament for treating and / or preventing a disease or disorder associated with TLR2 receptor.

23. A kit comprising the following components: - a compound as claimed in any one of claims 1 to 13; and optionally - a written description describing the use of the compound as claimed in any one of claims 15 to 22.

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