Methods and compositions for treatment of ophthalmic diseases

Abstract

Provided herein are compositions and methods of use for ophthalmic administration.

Description

[0001] Cross-reference

[0002] This application claims the benefit of U.S. Application No. 63 / 380,332, filed Oct. 20, 2022, which is hereby incorporated by reference in its entirety. Background of the Invention

[0003] Delivering drugs to the human eye is an important area of drug development. The anatomy and physiology of the eye pose challenges for drugs to reach the site of action and maintain therapeutic efficacy over time. Summary of the Invention

[0004] In one aspect, provided herein are compositions comprising a double-stranded DNA-binding moiety that binds to double-stranded deoxyribonucleic acid (DNA) in a sequence-specific manner. In some embodiments, the double-stranded DNA-binding moiety does not bind to a repetitive sequence comprising multiple copies of GAA. In some embodiments, the compositions are suitable for administration to the human eye.

[0005] In another aspect, provided herein are compositions comprising a double-stranded DNA-binding moiety suitable for treating genetic diseases, wherein the compositions are suitable for administration to the human eye.

[0006] In a further aspect, provided herein are compositions comprising a double-stranded DNA-binding moiety that binds to a repetitive sequence comprising at least four nucleotides, wherein the compositions are suitable for administration to the human eye.

[0007] A composition comprising a double-stranded DNA-binding moiety that binds to a repetitive sequence comprising multiple copies of: CTG, TGC, CAG, GGC, TCCAA, GGGGCC, CGG, GCC, GCG, ATTCT, TGGAA, GGCCTG, CCCCGCCCCGCG, or CCTG, wherein the composition is suitable for administration to the human eye.

[0008] A composition comprising a double-stranded DNA-binding moiety, wherein the double-stranded DNA-binding moiety is a polymer that does not comprise nucleotides or amino acids, and wherein the composition is suitable for administration to the human eye.

[0009] In various aspects of the compositions provided herein, in some embodiments, the double-stranded DNA binding moiety binds to a repetitive sequence comprising multiple copies of CTG or CAG. In some embodiments, the double-stranded DNA binding moiety binds to the minor groove of double-stranded DNA. In some embodiments, the double-stranded DNA binding moiety binds to a repetitive sequence adjacent to or within a gene. In some embodiments, the double-stranded DNA binding moiety modulates gene expression. In some embodiments, the gene is transcription factor 4 (TCF4). In some embodiments, the double-stranded DNA binding moiety binds non-covalently. In some embodiments, the double-stranded DNA binding moiety does not comprise a polynucleotide. In some embodiments, the double-stranded DNA binding moiety does not comprise a polypeptide. In some embodiments, the molecular weight of the double-stranded DNA binding moiety is less than about 10 kDa, about 7.5 kDa, about 5 kDa, or about 2.5 kDa. In some embodiments, the double-stranded DNA binding moiety comprises a polyamide. In some embodiments, the double-stranded DNA binding moiety further comprises a transcriptional regulator binding moiety. In some embodiments, the double-stranded DNA binding moiety is linked to the transcriptional regulator binding moiety via a linker. In some embodiments, administration of the composition to the eye results in minimal systemic exposure. In some embodiments, the composition further comprises an excipient. In some embodiments, the ophthalmic formulation composition has a pH of about 5 to about 8. In some embodiments, the composition has a viscosity of about 1 to about 50,000 cps at about 20°C.

[0010] In another aspect, provided herein is a method of treating an eye disorder in a human subject in need thereof, the method comprising administering a double-stranded DNA binding moiety that binds to double-stranded deoxyribonucleic acid (DNA) in a sequence-specific manner. In some embodiments, the double-stranded DNA binding moiety does not bind to a repetitive sequence comprising multiple copies of GAA.

[0011] In another aspect, provided herein is a method of treating an eye disorder in a human subject in need thereof, the method comprising administering a double-stranded DNA binding moiety, wherein the eye disorder is a genetic disease.

[0012] In a further aspect, provided herein is a method of treating an eye disorder in a human subject in need thereof, the method comprising administering a double-stranded DNA binding moiety to the eye of the subject, wherein the administration results in minimal systemic exposure of the double-stranded DNA binding moiety, and wherein the double-stranded DNA binding moiety binds to a repetitive sequence comprising: CTG, TGC, CAG, GGC, TCCAA, GGGGCC, CGG, GCC, GCG, GCT, GCA, GAC, AAGGG, ATTCT, TGGAA, GGCCTG, CCCCGCCCCGCG, or CCTG.

[0013] In another aspect, provided herein is a method of treating an eye disorder in a human subject in need thereof, the method comprising administering a double-stranded DNA binding moiety that binds to a repeat sequence comprising at least four nucleotides.

[0014] In a further aspect, provided herein is a method of treating an eye disorder in a human subject in need thereof, the method comprising administering a double-stranded DNA binding moiety, wherein the double-stranded DNA binding moiety is a polymer that does not contain nucleotides or amino acids.

[0015] In various aspects of the methods herein, in some embodiments, the double-stranded DNA binding moiety binds to a repeat sequence comprising multiple copies of CTG or CAG. In some embodiments, the double-stranded DNA binding moiety binds to the minor groove of double-stranded DNA. In some embodiments, the double-stranded DNA binding moiety binds to a repeat sequence adjacent to or within a gene. In some embodiments, the double-stranded DNA binding moiety modulates gene expression. In some embodiments, the gene is transcription factor 4 (TCF4). In some embodiments, the double-stranded DNA binding moiety binds non-covalently. In some embodiments, the double-stranded DNA binding moiety does not contain polynucleotides. In some embodiments, the double-stranded DNA binding moiety does not contain polypeptides. In some embodiments, the molecular weight of the double-stranded DNA binding moiety is less than about 10 kDa, about 7.5 kDa, about 5 kDa, or about 2.5 kDa. In some embodiments, the double-stranded DNA binding moiety comprises a polyamide. In some embodiments, the double-stranded DNA binding moiety further comprises a transcriptional modulator binding moiety. In some embodiments, the double-stranded DNA binding moiety is linked to the transcriptional modulator binding moiety via a linker. In some embodiments, the double-stranded DNA binding moiety is formulated in an excipient suitable for administration to the eye. In some embodiments, the double-stranded DNA binding moiety is in a formulation having a pH of about 5 to about 8. In some embodiments, the double-stranded DNA binding moiety is in a formulation having a viscosity of about 1 to about 50,000 cps at about 20°C.

[0016] Incorporation by reference

[0017] All publications, patents, and patent applications mentioned in this specification are incorporated herein by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] The features and advantages of the present invention will be understood by reference to the following detailed description of illustrative embodiments in which the principles of the invention are utilized, and in its accompanying drawings:

[0019] Figure 1 Showing the in vitro bovine eye distribution of compound 201. DETAILED DESCRIPTION

[0020] In treating diseases affecting the eye, it is necessary to treat the affected cells and tissues of the eye, while systemic administration and exposure are unnecessary or undesirable. Thus, in many cases, it is beneficial to directly administer treatment to the eye disease of the eye and minimize exposure to the rest of the body. In some cases, such features of eye disease treatment avoid off-target effects, reduce the immune response to the pharmaceutical composition, and improve therapeutic efficacy by ensuring that the maximum dose reaches the target cells and tissues of the eye.

[0021] In particular, some diseases of the eye are treated with compositions containing agents that bind to double-stranded deoxyribonucleic acid (DNA), as further described elsewhere herein. It has not been previously demonstrated that the direct administration of such compositions to the eye is accompanied by minimal systemic exposure, and thus the ophthalmic administration of these compositions has not been demonstrated in the art.

[0022] Compositions for ophthalmic administration are provided herein, such as ophthalmic compositions containing agents formulated for administration to the eye, and methods of treating eye diseases using such compositions. In some embodiments, the compositions provided contain agents that act by modulating gene expression by binding to double-stranded deoxyribonucleic acid (DNA). In some embodiments, such compositions contain agents that bind to the sequence and recruit transcriptional regulators (such as transcriptional activators or blockers), thereby modulating the expression of one or more genes and thus modulating genes adjacent to a particular DNA sequence. In additional embodiments, the administration of such compositions to the eye results in therapeutic retention in the eye and provides minimal contact with the rest of the body.

[0023] Double-stranded deoxyribonucleic acid binding composition

[0024] The present disclosure provides compositions comprising a double-stranded deoxyribonucleic acid (DNA) binding moiety. In some embodiments, the double-stranded DNA binding moiety binds to a repetitive sequence. In some embodiments, the repetitive sequence does not contain multiple copies of GAA. In some embodiments, the repetitive sequence comprises a repeat of at least four nucleotides. In some embodiments, the repetitive sequence comprises multiple copies of CTG, TGC, CAG, GGC, TCCAA, GGGGCC, CGG, GCC, GCG, GCT, GCA, GAC, AAGGG, ATTCT, TGGAA, GGCCTG, CCCCGCCCCGCG, or CCTG. In some embodiments, the repetitive sequence comprises multiple copies of CTG or CAG. In some embodiments, the repetitive sequence is not a triplet repeat. In some embodiments, the double-stranded DNA binding moiety is suitable for treating genetic diseases. In some embodiments, the genetic disease is not Friedreich's ataxia. In some embodiments, the double-stranded DNA binding moiety is a polymer that does not contain nucleotides or amino acids. In some embodiments, the double-stranded DNA binding moiety is not an antibiotic. In some embodiments, the composition is suitable for administration to the human eye. In some embodiments, the double-stranded DNA binding moiety binds to the minor groove of double-stranded DNA.

[0025] In aspects of the compositions provided herein, the double-stranded DNA binding moiety binds to a sequence (such as a repetitive sequence) adjacent to or within a gene. In some embodiments, the double-stranded DNA binding moiety regulates the expression of the gene. In some embodiments, the double-stranded DNA binding moiety increases the expression of the gene. In some embodiments, the double-stranded DNA binding moiety decreases the expression of the gene. In some embodiments, the gene is transcription factor 4 (TCF4).

[0026] In aspects of the compositions provided herein, the double-stranded DNA binding moiety binds non-covalently to double-stranded DNA. In some embodiments, the double-stranded DNA binding moiety binds to double-stranded DNA using electrostatic forces (such as hydrogen bonds or van der Waals forces). In some embodiments, the double-stranded DNA binding moiety does not contain polynucleotides. In some embodiments, the double-stranded DNA binding moiety does not contain polypeptides.

[0027] In aspects of the compositions provided herein, the molecular weight of the double-stranded DNA binding moiety is less than about 15 kDa, less than about 12.5 kDa, less than about 10 kDa, less than about 9.5 kDa, less than about 9 kDa, less than about 8.5 kDa, less than about 8 kDa, less than about 7.5 kDa, less than about 7 kDa, less than about 6.5 kDa, less than about 6 kDa, less than about 5.5 kDa, less than about 5 kDa, less than about 4.5 kDa, less than about 4 kDa, less than about 3.5 kDa, less than about 3 kDa, less than about 2.5 kDa, less than about 2 kDa or less than about 1.5 kDa. In some embodiments, the molecular weight of the double-stranded DNA binding moiety is less than about 10 kDa, about 7.5 kDa, about 5 kDa or about 2.5 kDa.

[0028] In some embodiments, the double-stranded DNA binding moiety comprises a polyamide. In some embodiments, the double-stranded DNA binding moiety further comprises a transcriptional regulator binding moiety. In some embodiments, the double-stranded DNA binding moiety is linked to the transcriptional regulator binding moiety via a linker.

[0029] In various aspects, the composition is formulated for topical administration. In some embodiments, the composition further comprises an excipient. In some embodiments, the excipient includes an isotonicity regulator (such as sodium chloride), a buffer, a stabilizer, an antioxidant, a thickening agent, a solubility enhancer, a penetration enhancer, and / or a preservative. In some embodiments, the pH of the composition is from about 5 to about 8. In some embodiments, the viscosity of the composition at about 20 °C is from about 1 to about 50,000 centipoise (cps).

[0030] In various aspects, administration of the composition to the eye results in minimal systemic exposure. In some embodiments, topical ocular administration (e.g., ophthalmic administration) of the composition results in penetration of the double-stranded DNA binding moiety into the inner cells and tissues of the eye. In some embodiments, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 95% or at least about 99% of the double-stranded DNA binding moiety administered to the eye can penetrate the inner cells and tissues of the eye.

[0031] In another aspect, provided herein is a composition comprising a double-stranded DNA binding moiety, wherein the double-stranded DNA binding moiety is a polymer that does not contain nucleotides or amino acids, and wherein the composition is suitable for administration to the human eye.

[0032] In some embodiments of the compositions that do not contain nucleotides or amino acids, the double-stranded DNA-binding moiety binds to a repetitive sequence. In some embodiments, the repetitive sequence comprises multiple copies of CTG, TGC, CAG, GGC, TCCAA, GGG GCC, CGG, GCC, GCG, GCT, GCA, GAC, AAGGG, ATTCT, TGGAA, GGCCTG, CCCCCGCCCCGCG, or CCTG. In some embodiments, the repetitive sequence comprises multiple copies of CTG or CAG. In some embodiments, the repetitive sequence is not a triplet repeat. In some embodiments, the repetitive sequence comprises a repeat of at least four nucleotides. In some embodiments, the repetitive sequence does not contain GAA. In some embodiments, the double-stranded DNA-binding moiety binds to the minor groove of double-stranded DNA.

[0033] In aspects of the compositions that do not contain nucleotides or amino acids provided herein, the double-stranded DNA-binding moiety is suitable for treating genetic diseases. In some embodiments, the genetic disease is not Friedreich's ataxia. In some embodiments, the double-stranded DNA-binding moiety is not an antibiotic.

[0034] In aspects of the compositions that do not contain nucleotides or amino acids provided herein, the double-stranded DNA-binding moiety binds to a sequence (such as a repetitive sequence) adjacent to or within a gene. In some embodiments, the double-stranded DNA-binding moiety regulates gene expression. In some embodiments, the double-stranded DNA-binding moiety increases gene expression. In some embodiments, the double-stranded DNA-binding moiety decreases gene expression. In some embodiments, the gene is transcription factor 4 (TCF4).

[0035] In aspects of the compositions that do not contain nucleotides or amino acids provided herein, the double-stranded DNA-binding moiety binds non-covalently to double-stranded DNA. In some embodiments, the double-stranded DNA-binding moiety binds to double-stranded DNA using electrostatic forces (such as hydrogen bonds or van der Waals forces).

[0036] In aspects of the compositions that do not contain nucleotides or amino acids provided herein, the molecular weight of the double-stranded DNA-binding moiety is less than about 15 kDa, less than about 12.5 kDa, less than about 10 kDa, less than about 9.5 kDa, less than about 9 kDa, less than about 8.5 kDa, less than about 8 kDa, less than about 7.5 kDa, less than about 7 kDa, less than about 6.5 kDa, less than about 6 kDa, less than about 5.5 kDa, less than about 5 kDa, less than about 4.5 kDa, less than about 4 kDa, less than about 3.5 kDa, less than about 3 kDa, less than about 2.5 kDa, less than about 2 kDa, or less than about 1.5 kDa. In some embodiments, the molecular weight of the double-stranded DNA-binding moiety is less than about 10 kDa, about 7.5 kDa, about 5 kDa, or about 2.5 kDa.

[0037] In some embodiments of the composition that does not contain nucleotides or amino acids, the double-stranded DNA-binding moiety comprises a polyamide. In some embodiments, the double-stranded DNA-binding moiety further comprises a transcriptional regulator-binding moiety. In some embodiments, the double-stranded DNA-binding moiety is linked to the transcriptional regulator-binding moiety via a linker.

[0038] In various aspects, the composition that does not contain nucleotides or amino acids is suitable for topical administration. In some embodiments, the composition further comprises an excipient. In some embodiments, the excipient includes an isotonicity regulator (such as sodium chloride), a buffer, a stabilizer and an antioxidant, a thickening agent, a solubility enhancer, a penetration enhancer, and / or a preservative. In some embodiments, the pH of the composition is from about 5 to about 8. In some embodiments, the viscosity of the composition at about 20 °C is from about 1 to about 50,000 cps.

[0039] In various aspects, administration of the composition that does not contain nucleotides or amino acids to the eye results in minimal systemic exposure. In some embodiments, topical administration of the composition to the eye (e.g., ophthalmic administration) causes the double-stranded DNA-binding moiety to penetrate into the internal cells and tissues of the eye. In some embodiments, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or at least about 99% of the double-stranded DNA-binding moiety administered ophthalmically can penetrate the cells and tissues inside the eye.

[0040] In a further aspect, the provided composition suitable for treating genetic diseases comprises a double-stranded DNA-binding moiety. In some embodiments, the genetic disease is not Friedreich's ataxia. In some embodiments, the double-stranded DNA-binding moiety is not an antibiotic.

[0041] In some embodiments of the composition suitable for treating genetic diseases, the composition comprises a double-stranded DNA-binding moiety that binds to a repeat sequence. In some embodiments, the repeat sequence does not contain multiple copies of GAA. In some embodiments, the repeat sequence contains multiple copies of CTG, TGC, CAG, GGC, TCCAA, GGGGCC, CGG, GCC, GCG, GCT, GCA, GAC, AAGGG, ATTCT, TGGAA, GGCCTG, CCCCGCCCCGCG, or CCTG. In some embodiments, the repeat sequence contains multiple copies of CTG or CAG. In some embodiments, the repeat sequence contains a repeat of at least four nucleotides. In some embodiments, the double-stranded DNA-binding moiety binds to the minor groove of double-stranded DNA.

[0042] In aspects of the compositions provided herein that are suitable for treating genetic diseases, the double-stranded DNA-binding moiety binds to a sequence adjacent to or within a gene. In some embodiments, the double-stranded DNA-binding moiety regulates gene expression. In some embodiments, the double-stranded DNA-binding moiety increases gene expression. In some embodiments, the double-stranded DNA-binding moiety decreases gene expression. In some embodiments, the gene is transcription factor 4 (TCF4).

[0043] In aspects of the compositions provided herein that are suitable for treating genetic diseases, the double-stranded DNA-binding moiety non-covalently binds to double-stranded DNA. In some embodiments, the double-stranded DNA-binding moiety binds to double-stranded DNA using electrostatic forces such as hydrogen bonds or van der Waals forces. In some embodiments, the double-stranded DNA-binding moiety does not comprise a polynucleotide. In some embodiments, the double-stranded DNA-binding moiety does not comprise a polypeptide.

[0044] In aspects of the compositions provided herein that are suitable for treating genetic diseases, the molecular weight of the double-stranded DNA-binding moiety is less than about 15 kDa, less than about 12.5 kDa, less than about 10 kDa, less than about 9.5 kDa, less than about 9 kDa, less than about 8.5 kDa, less than about 8 kDa, less than about 7.5 kDa, less than about 7 kDa, less than about 6.5 kDa, less than about 6 kDa, less than about 5.5 kDa, less than about 5 kDa, less than about 4.5 kDa, less than about 4 kDa, less than about 3.5 kDa, less than about 3 kDa, less than about 2.5 kDa, less than about 2 kDa or less than about 1.5 kDa. In some embodiments, the molecular weight of the double-stranded DNA-binding moiety is less than about 10 kDa, about 7.5 kDa, about 5 kDa or about 2.5 kDa.

[0045] In another aspect of the compositions provided herein that are suitable for treating genetic diseases, the composition comprises a double-stranded DNA-binding moiety, wherein the double-stranded DNA-binding moiety is a polymer that does not comprise nucleotides or amino acids.

[0046] In some embodiments of the compositions suitable for treating genetic diseases, the double-stranded DNA-binding moiety comprises a polyamide. In some embodiments, the double-stranded DNA-binding moiety further comprises a transcriptional regulator-binding moiety. In some embodiments, the double-stranded DNA-binding moiety is linked to the transcriptional regulator-binding moiety via a linker.

[0047] DNA-binding moiety

[0048] In aspects of the compositions provided herein that are suitable for treating genetic diseases, the double-stranded DNA-binding moiety has the structure of formula (A-1) or a pharmaceutically acceptable salt thereof:

[0049]

[0050] Wherein:

[0051] Each X 1 、X 2 、X 3 、X 4 、X 5 、X 6 、X 7 and X 8 is independently O or NR 2 ;

[0052] Each Y 1 、Y 2 、Y 3 、Y 4 、Y 5 、Y 6 、Y 7 and Y 8 is independently CH or N;

[0053] W 1 is hydrogen, halogen, optionally substituted C1-C 10 alkyl, -N(R 1e )2, -NR 1e C(O)R 1f 、-C(O)NR 1e R 1f 、-N=C(N(R 1e )2) 2、 -NR 1e C(O)R 1f 、-OC(O)NR 1e R 1f 、AA 1-10 、-Z B -P(O)(OR 1e ) 2、 -Z B -(CH2) p3 -P(O)(OR 1e )2 or -Z B -(CH2) p3 -O-P(O)(OR 1e )2, wherein

[0054] Each R 1e is independently hydrogen, halogen, optionally substituted C1-C 20 alkyl, optionally substituted C1-C 20 heteroalkyl or PEG 1-20 ;

[0055] Each R 1f is independently hydrogen, optionally substituted C1-C 20Alkyl, optionally substituted C1-C 20 heteroalkyl, optionally substituted 5-membered heteroaryl, PEG 1-20 or AA 1-10 ;

[0056] Each AA is independently a naturally occurring amino acid;

[0057] Z B is N or O; and

[0058] p3 is from 1 to 10;

[0059] W 2 is hydrogen, optionally substituted C1-C 20 alkyl or optionally substituted C1-C 20 heteroalkyl; or W 2 is -L 1 -Z-R 4 where

[0060] L 1 is alkylene or heteroalkylene;

[0061] Z is absent, or is -C(O)- or -C(=NH)-; and

[0062] R 4 is C1-C6 alkyl, -OR 4b or -NR 4a R 4b ; where

[0063] R 4a is hydrogen, optionally substituted C1-C 20 alkyl or optionally substituted C1-C 20 heteroalkyl;

[0064] R 4b is optionally substituted C1-C 20 alkyl, optionally substituted C2-C 20 alkenyl, optionally substituted C2-C 20 alkynyl, optionally substituted C1-C 20 aminoalkyl, optionally substituted C1-

[0065] C 20 haloalkyl, optionally substituted C1-C 20 heteroalkyl, optionally substituted C1-C 20 hydroxyalkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted 4- to 8-membered heterocycloalkyl, optionally substituted phenyl or optionally substituted 5- to 10-membered heteroaryl; or

[0066] R4a and R 4b together with the nitrogen to which they are attached form an optionally substituted 4- to 8-membered heterocycloalkyl group which is partially unsaturated or fully unsaturated;

[0067] R W is hydrogen or optionally substituted C1-C 20 alkyl; or

[0068] W 2 and R W together with the nitrogen to which they are attached form an optionally substituted 4- to 8-membered heterocycloalkyl group which is partially unsaturated or fully unsaturated;

[0069] Each R 2 independently is hydrogen, optionally substituted C1-C 20 alkyl, optionally substituted C2-C 20 alkenyl, optionally substituted C2-C 20 alkynyl, optionally substituted C1-C 20 heteroalkyl, optionally substituted C2-C 20 heteroalkenyl, optionally substituted C2-C 50 heteroalkynyl, optionally substituted C1-C 20 hydroxyalkyl, optionally substituted C1-C 20 aminoalkyl, optionally substituted C1-C 20 haloalkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted 3- to 8-membered heterocycloalkyl or optionally substituted PEG 1-20 ;

[0070] Each R 3 independently is hydrogen, halogen, acetyl, amino, amido, hydroxy, optionally substituted C1-C 20 alkyl, optionally substituted C1-C 20 haloalkyl, optionally substituted C1-C 20 alkylamino or optionally substituted C1-C 20 hydroxyalkyl; or

[0071] Two Rs 3 together with the atoms to which they are attached form a C3-C6 cycloalkyl or 4- to 6-membered heterocycloalkyl;

[0072] p1 is 3 or 4;

[0073] n1 and n2 are each independently 0 or 1;

[0074] n3 is 0, 1 or 2;

[0075] m1 is 0, 1, 2 or 3; and

[0076] n0 is 0 or 1, where neither n0 nor m1 is 0.

[0077] In some embodiments of formula (A-1), n0 is 1. In some embodiments of formula (A-1), n0 is 0.

[0078] In some embodiments of formula (A-1), n2 is 1. In some embodiments of formula (A-1), n2 is 0.

[0079] In some embodiments of formula (A-1), n3 is 2. In some embodiments of formula (A-1), n3 is 1. In some embodiments of formula (A-1), n3 is 0.

[0080] In some embodiments of formula (A-1), p1 is 3. In some embodiments of formula (A-1), p1 is 4.

[0081] In some embodiments of formula (A-1), each X 1 , X 2 , X 3 , X 4 , X 5 , X 6 , X 7 and X 8 is independently NR 2 .

[0082] In some embodiments of formula (A-1), each R 2 is independently hydrogen or optionally substituted C1-C 20 alkyl, where each alkyl is optionally substituted by one or more amino, amido, azido, cyano, ester, oxo(=O), urea, optionally substituted aryl or optionally substituted 5- to 10-membered heteroaryl. In some embodiments of formula (A-1), each R 2 is independently optionally substituted C1-C 10 alkyl. In some embodiments of formula (A-1), each R 2 is independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, pentyl or hexyl. In some embodiments of formula (A-1), each R 2 is methyl. In some embodiments of formula (A-1), each R 2 is hydrogen.

[0083] In some embodiments of formula (A-1), each R 3 is independently hydrogen, amino or amido. In some embodiments of formula (A-1), each R 3 is independently amino. In some embodiments of formula (A-1), each R3 is independently an amide group. In some embodiments of formula (A-1), each R 3 is hydrogen.

[0084] In some embodiments of formula (A-1), two Rs 3 together with the atoms to which they are attached form a C3-C6 cycloalkyl or a 4- to 6-membered heteroalkyl. In some embodiments of formula (A-1), two Rs 3 together with the atoms to which they are attached form a C3-C6 cycloalkyl. In some embodiments of formula (A-1), two Rs 3 together with the atoms to which they are attached form a 4- to 6-membered heteroalkyl.

[0085] In some embodiments of formula (A-1), W 2 is -L 1 -Z-R 4 .

[0086] In some embodiments of formula (A-1), R W is hydrogen.

[0087] In various aspects of the compositions provided herein that are suitable for the treatment of genetic diseases, the double-stranded DNA-binding moiety has the structure of formula (A-2) or a pharmaceutically acceptable salt thereof:

[0088]

[0089] Wherein:

[0090] W 1 is hydrogen or -N=C(N(R 1e 2)2, where each R 1e is independently hydrogen or a C1-C3 alkyl;

[0091] Each Y 1 , Y 2 , Y 3 , Y 4 , Y 5 , Y 6 , Y 7 and Y 8 are independently N or CH;

[0092] L 1 is a C1-C 20 alkylene or a C2-C 20 heteroalkylene;

[0093] Z is absent, or is -C(O)- or -C(=NH)-;

[0094] R 4 is a C1-C6 alkyl, -OR 4bor -NR 4a R 4b ; wherein

[0095] R 4a is hydrogen, optionally substituted C1 - C 20 alkyl or optionally substituted C1 - C 20 heteroalkyl;

[0096] R 4b is optionally substituted C1 - C 20 alkyl, optionally substituted C2 - C 20 alkenyl, optionally substituted C2 - C 20 alkynyl, optionally substituted C1 - C 20 aminoalkyl, optionally substituted C1 - C 20 haloalkyl, optionally substituted C1 - C 20 heteroalkyl, optionally substituted C1 - C 20 hydroxyalkyl, optionally substituted C3 - C8 cycloalkyl, optionally substituted 4 - to 8 - membered heterocycloalkyl, optionally substituted phenyl or optionally substituted 5 - to 10 - membered heteroaryl; or

[0097] R 4a and R 4b together with the nitrogen to which they are attached form an optionally substituted 4 - to 8 - membered heterocycloalkyl which is partially unsaturated or fully unsaturated;

[0098] Each R 2a 、R 2b 、R 2c 、R 2d 、R 2e 、R 2f 、R 2g and R 2h independently is hydrogen, optionally substituted C1 - C 20 alk

[0099] yl, optionally substituted C2 - C 20 alkenyl, optionally substituted C2 - C 20 alkynyl, optionally substituted C1 - C 20 heteroalkyl, optionally substituted C2 - C 20 heteroalkenyl, optionally substituted C2 - C 50 heteroalkynyl, optionally substituted C1 - C 20 hydroxyalkyl, optionally substituted C1 - C 20 aminoalkyl, optionally substituted C1 - C 20 haloalkyl, optionally substituted C3 - C8 cycloalkyl, optionally substituted 3 - to 8 - membered heterocycloalkyl

[0100] or optionally substituted PEG 1-20 ; wherein each group is optionally substituted with one or more R X substituents;

[0101] Each R 3a and R 3b are independently hydrogen, halogen, C1-C 10 alkyl, -OR 3e , -NR 3c R 3d or -NHC(O)R 3e , wherein

[0102] R 3c and R 3d are each independently hydrogen, alkyl or PEG;

[0103] R 3e is alkyl, PEG, cycloalkyl, heterocycloalkyl or phenyl;

[0104] Or two R 3a or two R 3b together with the carbon atom to which they are attached form a C3-C6 cycloalkyl or a 4- to 6-membered heterocycloalkyl;

[0105] Each R X is independently CN, -OH, -OR Xa , -N3, -NR Xa R Xb , -CO(O)R Xc , -C(O)OR Xc , -C(O)NR Xa R Xb ,

[0106] -NHC(O)R Xc , -NHC(O)OR Xc , -OC(O)NR Xa R Xb or optionally substituted 5- to 10-membered heteroaryl; wherein

[0107] R Xa and R Xb are each independently hydrogen, alkyl or PEG;

[0108] R Xc is alkyl, PEG, cycloalkyl, heterocycloalkyl or phenyl; and

[0109] n1 and m1 are each independently 0 or 1.

[0110] In some embodiments of formula (A-1) or (A-2), each Y 2 , Y4 , Y 7 and Y 8 is independently N. In some embodiments of formula (A-1) or (A-2), each Y 1 , Y 3 and Y 6 is independently CH.

[0111] In various aspects of the compositions provided herein that are suitable for the treatment of genetic diseases, the double-stranded DNA-binding moiety has the structure of formula (A-3) or a pharmaceutically acceptable salt thereof:

[0112]

[0113] Wherein:

[0114] W 1 is hydrogen or -N=C(N(R 1e ))2)2, wherein each R 1e is independently hydrogen or C1-C3 alkyl;

[0115] Each Y 5 is CH or N;

[0116] L 1 is C1-C 20 alkylene or C2-C 20 heteroalkylene;

[0117] Z is absent, or is -C(O)- or -C(=NH)-;

[0118] R 4 is C1-C6 alkyl, -OR 4b or -NR 4a R 4b ; wherein

[0119] R 4a is hydrogen, optionally substituted C1-C 20 alkyl or optionally substituted C1-C 20 heteroalkyl;

[0120] R 4b is optionally substituted C1-C 20 alkyl, optionally substituted C2-C 20 alkenyl, optionally substituted C2-C 20 alkynyl, optionally substituted C1-C 20 aminoalkyl, optionally substituted C1-C 20 haloalkyl, optionally substituted C1-C 20 heteroalkyl, optionally substituted C1-C 20a hydroxyalkyl group, an optionally substituted C3-C7 cycloalkyl group, an optionally substituted 4- to 8-membered heterocycloalkyl group, an optionally substituted phenyl group or an optionally substituted 5- to 10-membered heteroaryl group; or

[0121] R 4a and R 4b together with the nitrogen to which they are attached form an optionally substituted 4- to 8-membered heterocycloalkyl group which is partially unsaturated or fully unsaturated;

[0122] each R 2a 、R 2b 、R 2c 、R 2d 、R 2e 、R 2f 、R 2g and R 2h independently is hydrogen, an optionally substituted C1-C 20 alkyl

[0123] group, an optionally substituted C2-C 20 alkenyl group, an optionally substituted C2-C 20 alkynyl group, an optionally substituted C1-C 20 heteroalkyl group, an optionally substituted C2-C 20 heteroalkenyl group, an optionally substituted C2-C 50 heteroalkynyl group, an optionally substituted C1-C 20 hydroxyalkyl group, an optionally substituted C1-C 20 aminoalkyl group, an optionally substituted C1-C 20 haloalkyl group, an optionally substituted C3-C8 cycloalkyl group, an optionally substituted 3- to 8-membered heterocycloalkyl group or an optionally substituted PEG 1-20 ; wherein each group is optionally substituted by one or more R X substituents;

[0124] each R 3a and R 3b independently is hydrogen, a halogen, a C1-C 10 alkyl group, -OR 3e 、-NR 3c R 3d or -NHC(O)R 3e , wherein

[0125] R 3c and R 3d are each independently hydrogen, an alkyl group or a PEG;

[0126] R 3e is an alkyl group, a PEG, a cycloalkyl group, a heterocycloalkyl group or a phenyl group;

[0127] Or two Rs 3a Or two Rs 3b Together with the carbon atom to which they are attached form a C3-C6 cycloalkyl or a 4- to 6-membered heteroalkyl;

[0128] Each R X Independently is CN, -OH, -OR Xa , -N3, -NR Xa R Xb , -CO(O)R Xc , -C(O)OR Xc , -C(O)NR Xa R Xb ,

[0129] -NHC(O)R Xc , -NHC(O)OR Xc , -OC(O)NR Xa R Xb Or an optionally substituted 5- to 10-membered heteroaryl; wherein

[0130] R Xa and R Xb Each independently is hydrogen, alkyl or PEG;

[0131] R Xc Is alkyl, PEG, cycloalkyl, heteroalkyl or phenyl; and

[0132] n1 and m1 each independently is 0 or 1.

[0133] In various aspects of the compositions provided herein that are suitable for the treatment of genetic diseases, the double-stranded DNA-binding moiety has the structure of formula (A-4) or a pharmaceutically acceptable salt thereof:

[0134]

[0135] Wherein:

[0136] W 1 Is hydrogen or -N = C(N(R 1e ))2)2, wherein each R 1e Independently is hydrogen or C1-C3 alkyl;

[0137] Each Y 1 , Y 2 , Y 3 , Y 4 , Y 5 , Y 6 , Y 7 and Y 8 Independently is N or CH;

[0138] L Vis C1-C 20 alkylene, C2-C 20 heteroalkylene or AA 1-10 ; wherein each AA is independently a naturally occurring amino acid;

[0139] V is absent or is optionally substituted C3-C8 cycloalkyl, optionally substituted 4- to 8-membered heteroalkyl, optionally substituted phenyl or optionally substituted 5- to 10-membered heteroaryl;

[0140] Each R 2a , R 2b , R 2c , R 2d , R 2e , R 2f , R 2g and R 2h is independently hydrogen, optionally substituted C1-C 20 alkane

[0141] yl, optionally substituted C2-C 20 alkenyl, optionally substituted C2-C 20 alkynyl, optionally substituted C1-C 20 heteroalkyl, optionally substituted C2-C 20 heteroalkenyl, optionally substituted C2-C 50 heteroalkynyl, optionally substituted C1-C 20 hydroxyalkyl, optionally substituted C1-C 20 aminoalkyl, optionally substituted C1-C 20 haloalkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted 3- to 8-membered heteroalkyl or optionally substituted PEG 1-20 ; wherein each group is optionally substituted by one or more R X substituents;

[0142] Each R 3a and R 3b is independently hydrogen, halogen, C1-C 10 alkyl, -OR 3e , -NR 3c R 3d or -NHC(O)R 3e , wherein

[0143] R 3c and R 3d are each independently hydrogen, alkyl or PEG;

[0144] R 3e is alkyl, PEG, cycloalkyl, heteroalkyl or phenyl;

[0145] or two Rs 3a or two Rs 3b together with the carbon atom to which they are attached form a C3-C6 cycloalkyl or a 4- to 6-membered heteroalkyl;

[0146] Each R X is independently CN, -OH, -OR Xa , -N3, -NR Xa R Xb , -CO(O)R Xc , -C(O)OR Xc , -C(O)NR Xa R Xb ,

[0147] , -NHC(O)R Xc , -NHC(O)OR Xc , -OC(O)NR Xa R Xb or an optionally substituted 5- to 10-membered heteroaryl; wherein

[0148] R Xa and R Xb are each independently hydrogen, alkyl or PEG;

[0149] R Xc is alkyl, PEG, cycloalkyl, heteroalkyl or phenyl;

[0150] n1 and m1 are each independently 0 or 1; and

[0151] xv is from 0 to 10.

[0152] In some embodiments of formula (A-4), each Y 2 , Y 4 , Y 7 and Y 8 is independently N. In some embodiments of formula (A-4), each Y 1 , Y 3 and Y 6 is independently CH.

[0153] In some embodiments of formula (A-4), L V is C1-C 20 alkylene or C2-C 20 heteroalkylene. In some embodiments of formula (A-4), L V is C1-C 20 alkylene. In some embodiments of formula (A-4), L V is C1-C 10 alkylene. In some embodiments of formula (A-4), L Vis C2-C 20 heteroalkylene. In some embodiments of formula (A-4), L V is C2-C 10 heteroalkylene. In some embodiments of formula (A-4), L V is PEG 1-10 .

[0154] In some embodiments of formula (A-4), xv is from 0 to 8, from 0 to 6, from 0 to 4 or from 0 to 2. In some embodiments of formula (A-4), xv is from 0 to 8. In some embodiments of formula (A-4), xv is from 0 to 4. In some embodiments of formula (A-4), xv is 0, 1, 2 or 3.

[0155] In some embodiments, V is optionally substituted C3-C8 cycloalkyl or optionally substituted 4- to 8-membered heteroalkyl. In some embodiments, V is optionally substituted C3-C8 cycloalkyl. In some embodiments, V is optionally substituted 4- to 8-membered heteroalkyl. In some embodiments, V is optionally substituted 6-membered heteroalkyl. In some embodiments, V is optionally substituted phenyl or optionally substituted 5- to 10-membered heteroaryl. In some embodiments, V is optionally substituted phenyl. In some embodiments, V is optionally substituted 5- to 10-membered heteroaryl. In some embodiments, V is absent.

[0156] In some embodiments of formula (A-4), V has the structure of formula (C-1) or a pharmaceutically acceptable salt thereof:

[0157]

[0158] Wherein:

[0159] B 1 is -CR 60 R 60 -, -O-, -NR 60 -, -S(O)-, -S(O)2- or -S-; or B 1 is where each R 60 is independently hydrogen, optionally substituted C1-C 20 alkyl, optionally substituted C2-C 20 alkenyl, optionally substituted C2-C 20 alkynyl, optionally substituted C1-C 20 heteroalkyl, -C(O)OR 60a or -

[0160] C(O)R 60a , where R 60a is hydrogen, optionally substituted C1-C20 alkyl, optionally substituted C1-

[0161] C 10 haloalkyl, optionally substituted PEG 1-20 , optionally substituted C3-C6 cycloalkyl, optionally substituted 3- to 6-membered heterocycloalkyl or optionally substituted phenyl; or

[0162] two Rs 60 together with the nitrogen atom to which they are attached form an optionally substituted 4- to 8-membered heterocycloalkyl;

[0163] Ring P is optionally substituted C3-C6 cycloalkyl, optionally substituted 4- to 6-membered heterocycloalkyl, optionally substituted phenyl or optionally substituted 5- to 10-membered heteroaryl;

[0164] L b is absent, or is C1-C4 alkylene, C2-C4 alkenylene or C2-C4 alkynylene; and

[0165] a1 and a2 are each independently 0, 1 or 2.

[0166] In some embodiments of formula (C-1), B 1 is -CR 60 R 60 -, -O-, -NR 60 -, -S(O)-, -S(O)2- or -S-. In some embodiments of formula (C-1), B 1 is

[0167] In some embodiments of formula (C-1), each R 60 is independently hydrogen, optionally substituted C1-C 20 alkyl, optionally substituted C2-C 20 alkenyl, optionally substituted C2-C 20 alkynyl or optionally substituted C1-C 20 heteroalkyl. In some embodiments of formula (C-1), each R 60 is independently -C(O)OR 60a or -C(O)R 60a . In some embodiments of formula (C-1), each R 60 is hydrogen.

[0168] In some embodiments of formula (C-1), two Rs 60 together with the nitrogen atom to which they are attached form an optionally substituted 4- to 8-membered heterocycloalkyl.

[0169] In some embodiments of formula (A-4), V has the structure of formula (C-2) or a pharmaceutically acceptable salt thereof:

[0170]

[0171] Wherein:

[0172] Ring P is an optionally substituted C3-C6 cycloalkyl, an optionally substituted 4- to 6-membered heterocycloalkyl, an optionally substituted phenyl, or an optionally substituted 5- to 10-membered heteroaryl;

[0173] L b is absent or is a C1-C4 alkylene, a C2-C4 alkenylene, or a C2-C4 alkynylene;

[0174] B 1 ’ is CH or N; and

[0175] a1 and a2 are each independently 0, 1, or 2.

[0176] In some embodiments of formula (C-2), ring P is an optionally substituted C3-C6 cycloalkyl or an optionally substituted 4- to 6-membered heterocycloalkyl. In some embodiments of formula (C-2), ring P is an optionally substituted C3-C6 cycloalkyl. In some embodiments of formula (C-2), ring P is an optionally substituted 4- to 6-membered heterocycloalkyl. In some embodiments of formula (C-2), ring P is an optionally substituted 6-membered heterocycloalkyl.

[0177] In some embodiments of formula (C-1) or (C-2), a1 and a2 are each independently 0 or 1. In some embodiments of formula (C-1) or (C-2), a1 and a2 are each 1. In some embodiments of formula (C-1) or (C-2), a1 and a2 are each independently 0. In some embodiments of formula (C-1) or (C-2), a1 is 0 and a2 is 1.

[0178] In some embodiments of formula (A-4), V has the structure of formula (C-3) or a pharmaceutically acceptable salt thereof:

[0179]

[0180] Wherein:

[0181] B 1 ’ and B 2 are each independently CH or N; and

[0182] B 3 is -CR 61 R 61 -, -O-, -S-, -S(O)-, -S(O)2-, or -NR61 -; wherein

[0183] each R 61 is independently hydrogen, optionally substituted C1-C 20 alkyl, optionally substituted C2-C 20 alkenyl, optionally substituted C2-C 20 alkynyl, optionally substituted C1-C 20 heteroalkyl, -C(O)OR 61a or -

[0184] C(O)R 61a ; wherein R 61a is hydrogen, optionally substituted C1-C 20 alkyl, optionally substituted C1-

[0185] C 10 haloalkyl, optionally substituted PEG 1-20 , optionally substituted C3-C6 cycloalkyl, optionally substituted 3- to 6-membered heteroalkyl or optionally substituted phenyl; and

[0186] L b is absent or is C1-C4 alkylene, C2-C4 alkenylene or C2-C4 alkynylene.

[0187] In some embodiments of formula (C-2) or (C-3), B 1 ' is CH. In some embodiments of formula (C-2) or (C-3), B 1 ' is N.

[0188] In some embodiments of formula (C-1), (C-2) or (C-3), L b is C1-C4 alkylene, C2-C4 alkenylene or C2-C4 alkynylene. In some embodiments of formula (C-1), (C-2) or (C-3), L b is absent or is C2-C4 alkynylene. In some embodiments of formula (C-1), (C-2) or (C-3), L b is C2-C4 alkynylene. In some embodiments of formula (C-1), (C-2) or (C-3), L b is absent.

[0189] In some embodiments of formula (A-4), V has the structure of formula (C-4) or a pharmaceutically acceptable salt thereof

[0190]

[0191] wherein:

[0192] B2 is CH or N;

[0193] B 3 is -CR 61 R 61 -, -O-, -S-, -S(O)-, -S(O)2- or -NR 61 -; wherein

[0194] each R 61 is independently hydrogen, optionally substituted C1-C 20 alkyl, optionally substituted C2-C 20 alkenyl, optionally substituted C2-C 20 alkynyl, optionally substituted C1-C 20 heteroalkyl, -C(O)OR 61a or -

[0195] C(O)R 61a ; wherein R 61a is hydrogen, optionally substituted C1-C 20 alkyl, optionally substituted C1-

[0196] C 10 haloalkyl, optionally substituted PEG 1-20 , optionally substituted C3-C6 cycloalkyl, optionally substituted 3- to 6-membered heteroalkyl or optionally substituted phenyl; and

[0197] R 62a is hydrogen, optionally substituted C1-C 20 alkylene or optionally substituted PEG 1-20 ;

[0198] each R 62 is independently hydrogen or C1-C3 alkyl; and

[0199] s2 is 1, 2 or 3.

[0200] In some embodiments of formula (C-3) or (C-4), B 2 is CH. In some embodiments of formula (C-3) or (C-4), B 2 is N.

[0201] In some embodiments of formula (C-3) or (C-4), B 3 is -CR 61 R 61 -, -O- or -NR 61 -. In some embodiments of formula (C-3) or (C-4), B 3 is -CR 61 R 61-。In some embodiments of formula (C-3) or (C-4), B 3 is -O-. In some embodiments of formula (C-3) or (C-4), B 3 is -NR 61 -. In some embodiments of formula (C-3) or (C-4), B 3 is -S-, -S(O)- or -S(O)2-.

[0202] In some embodiments of formula (C-3) or (C-4), each R 61 is independently hydrogen, optionally substituted C1-C 20 alkyl, optionally substituted C2-C 20 alkenyl, optionally substituted C2-C 20 alkynyl or optionally substituted C1-C 20 heteroalkyl. In some embodiments of formula (C-3) or (C-4), each R 61 is independently -C(O)OR 61a or -C(O)R 61a . In some embodiments of formula (C-3) or (C-4), each R 61 is hydrogen.

[0203] In some embodiments of formula (C-3) or (C-4), R 61a is optionally substituted C1-C 20 alkyl. In some embodiments of formula (C-3) or (C-4), R 61a is optionally substituted phenyl. In some embodiments of formula (C-3) or (C-4), R 61a is optionally substituted PEG 1-20 . In some embodiments of formula (C-3) or (C-4), R 61a is hydrogen.

[0204] In some embodiments of formula (C-4), each R 62 is independently C1-C3 alkyl. In some embodiments of formula (C-4), each R 62 is hydrogen.

[0205] In some embodiments of formula (C-4), R 62a is optionally substituted C1-C 20 alkylene or optionally substituted PEG 1-20 . In some embodiments of formula (C-4), R 62a is hydrogen.

[0206] In some embodiments of formula (C-4), s2 is 1 or 2. In some embodiments of formula (C-4), s2 is 2. In some embodiments of formula (C-4), s2 is 1.

[0207] In some embodiments of formula (A-2), (A-3) or (A-4), each R 2a , R 2b , R 2c , R 2d , R 2e , R 2f , R 2g and R 2h is independently hydrogen, optionally substituted C1-C 20 alkyl, optionally substituted C1-C 20 heteroalkyl, optionally substituted C1-C 20 hydroxyalkyl, optionally substituted C1-C 20 aminoalkyl, optionally substituted C1-C 20 haloalkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted 3- to 8-membered heteroalkyl or optionally substituted PEG 1-20 ; wherein each group is optionally substituted by one or more R X . In some embodiments of formula (A-2), (A-3) or (A-4), each R 2a , R 2b , R 2c , R 2d , R 2e , R 2f , R 2g and R 2h is independently hydrogen, optionally substituted C1-C 20 alkyl, optionally substituted C1-C 20 haloalkyl, optionally substituted C1-C 20 heteroalkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted 4- to 8-membered heteroalkyl, wherein each group is optionally substituted by one or more R X . In some embodiments of formula (A-2), (A-3) or (A-4), each R 2a , R 2b , R 2c , R 2d , R 2e , R 2f , R 2g and R 2h is independently hydrogen, optionally substituted C1-C 20 alkyl, optionally substituted C1-C 20Haloalkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted 4- to 8-membered heterocycloalkyl, wherein each group is optionally substituted by one or more R X substituents. In some embodiments of formula (A-2), (A-3), or (A-4), each R 2a , R 2b , R 2c , R 2d , R 2e , R 2f , R 2g and R 2h is independently hydrogen or optionally substituted C1-C 20 alkyl, wherein each alkyl is optionally substituted by one or more R X substituents. In some embodiments of formula (A-2), (A-3), or (A-4), each R 2a , R 2b , R 2c , R 2d , R 2e , R 2f , R 2g and R 2h is independently optionally substituted C3-C8 cycloalkyl or optionally substituted 4- to 8-membered heterocycloalkyl. In some embodiments of formula (A-2), (A-3), or (A-4), each R 2a , R 2b , R 2c , R 2d , R 2e , R 2f , R 2g and R 2h is independently optionally substituted C3-C8 cycloalkyl. In some embodiments of formula (A-2), (A-3), or (A-4), each R 2a , R 2b , R 2c , R 2d , R 2e , R 2f , R 2g and R 2h is independently optionally substituted 4- to 8-membered heterocycloalkyl. In some embodiments of formula (A-2), (A-3), or (A-4), each R 2a , R 2b , R 2c , R 2d , R 2e , R 2f , R 2g and R 2h is independently optionally substituted C1-C 20Heteroalkyl. In some embodiments of formula (A-2), (A-3), or (A-4), each R 2a , R 2b , R 2c , R 2d , R 2e , R 2f , R 2g and R 2h is independently an optionally substituted C1-C 10 alkyl. In some embodiments of formula (A-2), (A-3), or (A-4), each R 2a , R 2b , R 2c , R 2d , R 2e , R 2f , R 2g and R 2h is independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, pentyl, or hexyl. In some embodiments of formula (A-2), (A-3), or (A-4), each R 2a , R 2b , R 2c , R 2d , R 2e , R 2f , R 2g and R 2h is methyl. In some embodiments of formula (A-2), (A-3), or (A-4), each R 2a , R 2b , R 2c , R 2d , R 2e , R 2f , R 2g and R 2h is hydrogen.

[0208] In some embodiments of formula (A-2), (A-3), or (A-4), each R 3a and R 3b is independently hydrogen, -NR 3c R 3d or -NHC(O)R 3f . In some embodiments of formula (A-2), (A-3), or (A-4), each R 3a and R 3b is independently hydrogen or -NR 3c R 3d . In some embodiments of formula (A-2), (A-3), or (A-4), each R 3a and R 3bIndependently is hydrogen or -NH2. In some embodiments of formula (A-2), (A-3), or (A-4), each R 3a and R 3b is hydrogen.

[0209] In some embodiments of formula (A-2), (A-3), or (A-4), two Rs 3a together with the carbon atom to which they are attached form a C3-C6 cycloalkyl. In some embodiments of formula (A-2), (A-3), or (A-4), two Rs 3a together with the carbon atom to which they are attached form cyclopropyl, cyclobutyl, or cyclopentyl. In some embodiments of formula (A-2), (A-3), or (A-4), two Rs 3a together with the carbon atom to which they are attached form cyclopropyl. In some embodiments of formula (A-2), (A-3), or (A-4), two Rs 3a together with the carbon atom to which they are attached form cyclobutyl. In some embodiments of formula (A-2), (A-3), or (A-4), two Rs 3a together with the carbon atom to which they are attached form cyclopentyl. In some embodiments of formula (A-2), (A-3), or (A-4), two Rs 3a together with the carbon atom to which they are attached form a 4- to 6-membered heterocycloalkyl.

[0210] In some embodiments of formula (A-2), (A-3), or (A-4), two Rs 3b together with the carbon atom to which they are attached form a C3-C6 cycloalkyl. In some embodiments of formula (A-2), (A-3), or (A-4), two Rs 3b together with the carbon atom to which they are attached form cyclopropyl, cyclobutyl, or cyclopentyl. In some embodiments of formula (A-2), (A-3), or (A-4), two Rs 3b together with the carbon atom to which they are attached form cyclopropyl. In some embodiments of formula (A-2), (A-3), or (A-4), two Rs 3b together with the carbon atom to which they are attached form cyclobutyl. In some embodiments of formula (A-2), (A-3), or (A-4), two Rs 3b together with the carbon atom to which they are attached form cyclopentyl. In some embodiments of formula (A-2), (A-3), or (A-4), two Rs 3b together with the carbon atom to which they are attached form a 4- to 6-membered heterocycloalkyl.

[0211] In some embodiments of formula (A-1), (A-2), or (A-3), L 1 is C1-C 10 alkylene or C2-C 10Heteroalkyl group. In some embodiments of formula (A-1), (A-2) or (A-3), L 1 is C1-C 10 alkylene. In some embodiments, L 1 is C2-C 10 heteroalkyl. In some embodiments of formula (A-1), (A-2) or (A-3), the heteroalkyl is polyethylene glycol. In some embodiments of formula (A-1), (A-2) or (A-3), L 1 is PEG 1-10 . In some embodiments of formula (A-1), (A-2) or (A-3), L 1 is –(CH2CH2-O) y1 -, where y1 is an integer in the range of 1 to 10. In some embodiments of formula (A-1), (A-2) or (A-3), the heteroalkyl group contains -(CH2) x3 N(R a )(CH2) x4 –, where R a is hydrogen or optionally substituted C1-C6 alkyl; and each x3 and x4 is independently an integer in the range of 1 to 6.

[0212] In some embodiments of formula (A-1), (A-2) or (A-3), Z is -C(O)-; and R 4 is -NR 4a R 4b . In some embodiments of formula (A-1), (A-2) or (A-3), Z is absent; and R 4 is -OR 4b . In some embodiments of formula (A-1), (A-2) or (A-3), Z is absent; and R 4 is -NR 4a R 4b . In some embodiments of formula (A-1), (A-2) or (A-3), Z is -C(O)-; and R 4 is optionally substituted C1-C6 alkyl. In some embodiments of formula (A-1), (A-2) or (A-3), Z is absent; and R 4 is optionally substituted C1-C6 alkyl.

[0213] In some embodiments of formula (A-1), (A-2) or (A-3), R 4a is hydrogen, optionally substituted C1-C 20 alkyl or optionally substituted C1-C 20 heteroalkyl. In some embodiments of formula (A-1), (A-2) or (A-3), R 4ais optionally substituted C1-C 20 alkyl or optionally substituted C1-C 20 heteroalkyl. In some embodiments of formula (A-1), (A-2) or (A-3), R 4a is optionally substituted C1-C 20 alkyl. In some embodiments of formula (A-1), (A-2) or (A-3), R 4a is optionally substituted C1-C 20 heteroalkyl. In some embodiments of formula (A-1), (A-2) or (A-3), the heteroalkyl is polyethylene glycol (PEG). In some embodiments of formula (A-1), (A-2) or (A-3), R 4a is optionally substituted PEG 1-20 . In some embodiments of formula (A-1), (A-2) or (A-3), R 4a is hydrogen.

[0214] In some embodiments of formula (A-1), (A-2) or (A-3), R 4b is hydrogen, optionally substituted C1-C 20 alkyl or optionally substituted C1-C 20 heteroalkyl. In some embodiments of formula (A-1), (A-2) or (A-3), R 4b is optionally substituted C1-C 20 alkyl or optionally substituted C1-C 20 heteroalkyl. In some embodiments of formula (A-1), (A-2) or (A-3), R 4b is optionally substituted C1-C 20 alkyl. In some embodiments of formula (A-1), (A-2) or (A-3), R 4b is optionally substituted C1-C 20 heteroalkyl. In some embodiments of formula (A-1), (A-2) or (A-3), the heteroalkyl is polyethylene glycol (PEG). In some embodiments of formula (A-1), (A-2) or (A-3), R 4b optionally substituted PEG 1-20 . In some embodiments of formula (A-1), (A-2) or (A-3), R 4b is hydrogen.

[0215] In some embodiments of formula (A-1), (A-2) or (A-3), R 4bis an optionally substituted C3-C8 cycloalkyl, an optionally substituted 4- to 8-membered heterocycloalkyl, an optionally substituted phenyl, or an optionally substituted 5- to 10-membered heteroaryl. In some embodiments of formula (A-1), (A-2), or (A-3), R 4b is an optionally substituted C3-C8 cycloalkyl or an optionally substituted 4- to 8-membered heterocycloalkyl. In some embodiments of formula (A-1), (A-2), or (A-3), R 4b is an optionally substituted C3-C6 cycloalkyl. In some embodiments of formula (A-1), (A-2), or (A-3), R 4b is an optionally substituted 4- to 6-membered heterocycloalkyl.

[0216] In some embodiments of formula (A-1), (A-2), or (A-3), R 4a and R 4b together with the nitrogen to which they are attached form an optionally substituted 4- to 8-membered heterocycloalkyl, which is partially unsaturated or fully unsaturated. In some embodiments of formula (A-1), (A-2), or (A-3), R 4a and R 4b together with the nitrogen to which they are attached form an optionally substituted 4- to 6-membered heterocycloalkyl. In some embodiments of formula (A-1), (A-2), or (A-3), R 4a and R 4b together with the nitrogen to which they are attached form an optionally substituted piperidine, piperazine, or morpholine.

[0217] In some embodiments of formula (A-2), (A-3), or (A-4), each R X is independently -CN, -OH, -OR Xa , -N3, -NR Xa R Xb , -C(O)OR Xc , -C(O)NR Xa R Xb or -NHC(O)R Xc . In some embodiments of formula (A-2), (A-3), or (A-4), each R X is independently -CN, -OH, -OR Xa , -N3 or -NR Xa R Xb . In some embodiments of formula (A-2), (A-3), or (A-4), each R X is independently -CO(O)R Xc , -C(O)OR Xc , -C(O)NR Xa R Xb or -NHC(O)RXc .

[0218] In some embodiments of formula (A-2), (A-3), or (A-4), each R Xa and R Xb is independently hydrogen, C1-C 20 alkyl, or PEG 1-20 . In some embodiments of formula (A-2), (A-3), or (A-4), each R Xa and R Xb is independently C1-C 20 alkyl. In some embodiments of formula (A-2), (A-3), or (A-4), each R Xa and R Xb is independently PEG 1-20 . In some embodiments of formula (A-2), (A-3), or (A-4), each R Xa and R Xb is hydrogen.

[0219] In some embodiments of formula (A-2), (A-3), or (A-4), R Xc is C1-C 20 alkyl, PEG 1-20 , C3-C6 cycloalkyl, 4- to 6-membered heteroalkyl, or phenyl. In some embodiments of formula (A-2), (A-3), or (A-4), R Xc is C1-C 20 alkyl or PEG 1-20 . In some embodiments of formula (A-2), (A-3), or (A-4), R Xc is C1-C 20 alkyl. In some embodiments of formula (A-2), (A-3), or (A-4), R Xc is PEG 1-20 .

[0220] In some embodiments of formula (A-2), (A-3), or (A-4), R 3c and R 3d are each independently hydrogen, C1-C 20 alkyl, or PEG 1-20 . In some embodiments of formula (A-2), (A-3), or (A-4), R 3c and R 3d are each independently hydrogen or C1-C 20 alkyl. In some embodiments of formula (A-2), (A-3), or (A-4), R 3c and R 3d are each independently hydrogen.

[0221] In some embodiments of formula (A-2), (A-3), or (A-4), R 3e is C1-C 20 alkyl, PEG 1-20 , C3-C6 cycloalkyl, 4- to 6-membered heteroalkyl, or phenyl. In some embodiments of formula (A-2), (A-3), or (A-4), R 3e is C1-C 20 alkyl or PEG 1-20 .

[0222] In some embodiments of formula (A-1), (A-2), (A-3), or (A-4), each Y 5 is independently N. In some embodiments of formula (A-1), (A-2), (A-3), or (A-4), each Y 5 is independently CH.

[0223] In some embodiments of formula (A-1), (A-2), (A-3), or (A-4), W 1 is -N=C(N(R 1e ))2)2, where each R 1e is independently hydrogen or C1-C3 alkyl. In some embodiments of formula (A-1), (A-2), (A-3), or (A-4), W 1 is hydrogen.

[0224] In some embodiments of formula (A-1), (A-2), (A-3), or (A-4), n1 is 1. In some embodiments of formula (A-1), (A-2), (A-3), or (A-4), n1 is 0.

[0225] In some embodiments of formula (A-1), (A-2), (A-3), or (A-4), m1 is 0 or 1. In some embodiments of formula (A-1), (A-2), (A-3), or (A-4), m1 is 1. In some embodiments of formula (A-1), (A-2), (A-3), or (A-4), m1 is 0.

[0226] In aspects of the compositions provided herein that are suitable for the treatment of genetic diseases, the double-stranded DNA-binding moiety is optionally linked to a second end comprising a moiety that regulates gene expression. In some embodiments, the double-stranded DNA-binding moiety is linked to the second end of the moiety that regulates gene expression by means of a linker.

[0227] In some embodiments, the second end of the portion comprising the gene expression modulating moiety is conjugated to a pyrrole on the double-stranded DNA binding moiety. In some embodiments, the second end of the portion comprising the gene expression modulating moiety is conjugated to an imidazole on the double-stranded DNA binding moiety. In some embodiments, the second end of the portion comprising the gene expression modulating moiety is conjugated to a β-alanine of the double-stranded DNA binding moiety. In some embodiments, the second end of the portion comprising the gene expression modulating moiety is conjugated to the C-terminus of the double-stranded DNA binding moiety. In some embodiments, the second end of the portion comprising the gene expression modulating moiety is conjugated to the N-terminus of the double-stranded DNA binding moiety. In some embodiments, the second end of the portion comprising the gene expression modulating moiety is conjugated to an amide of the double-stranded DNA binding moiety.

[0228] In aspects of the compositions provided herein that are suitable for the treatment of genetic diseases, the double-stranded DNA binding moiety of formula (A-1), (A-2), or (A-3) is optionally linked to the second end of the portion comprising the gene expression modulating moiety by way of an oligomeric linker. In aspects of the compositions provided herein that are suitable for the treatment of genetic diseases, the double-stranded DNA binding moiety of formula (A-1) is optionally linked to the second end of the portion comprising the gene expression modulating moiety. In aspects of the compositions provided herein that are suitable for the treatment of genetic diseases, the double-stranded DNA binding moiety of formula (A-2) is optionally linked to the second end of the portion comprising the gene expression modulating moiety. In aspects of the compositions provided herein that are suitable for the treatment of genetic diseases, the double-stranded DNA binding moiety of formula (A-3) is optionally linked to the second end of the portion comprising the gene expression modulating moiety.

[0229] In some embodiments, the second end of the portion comprising the gene expression modulating moiety is conjugated at W 1 . In some embodiments, the second end of the portion comprising the gene expression modulating moiety is conjugated at W 2 . In some embodiments, the second end of the portion comprising the gene expression modulating moiety is conjugated at one of R 2 . In some embodiments, the second end of the portion comprising the gene expression modulating moiety is conjugated at one of R 2a , R 2b , R 2c , R 2d , R 2e , R 2f , R 2g or R 2h . In some embodiments, the second end of the portion comprising the gene expression modulating moiety is conjugated at one of R 3 . In some embodiments, the second end of the portion comprising the gene expression modulating moiety is conjugated at one of R 3a . In some embodiments, the second end of the portion comprising the gene expression modulating moiety is conjugated at one of R 3bat one position. In some embodiments, the second end of the portion that regulates gene expression is conjugated at R 4 at one position. In some embodiments, the second end of the portion that regulates gene expression is conjugated at R 4a or R 4b at one position.

[0230] The portion that regulates gene expression

[0231] In some embodiments, the second end includes a portion that regulates gene expression.

[0232] In some embodiments, the second end includes a bromodomain-binding portion.

[0233] In some embodiments, the second end contains a portion capable of binding to a bromodomain and extra-terminal domain (BET) family member.

[0234] In some embodiments, the BET family member is BRD2, BRD3, BRD4, or BRDT. In some embodiments, the BET family member is BRD2. In some embodiments, the BET family member is BRD3. In some embodiments, the BET family member is BRD4. In some embodiments, the BET family member is BRDT.

[0235] In some embodiments, the protein-binding portion binds to CBP / p300, PCAF (P300 / CBP-associated factor), CECR2 (cat eye syndrome chromosomal candidate gene 2), BRPF (bromodomain and PHD finger-containing protein), ATAD2 / ATAD2B (chromatin remodeling protein), TRIM24 (tripartite motif-containing protein 24), BAZ2 (bromodomain adjacent to zinc finger domain), TAF1 (TBP-associated factor), BRD7 / 9, BPTF (bromodomain PHD finger transcription factor), SMARCA2 / 4, or PBRM1.

[0236] In some embodiments, the regulatory molecule is CBP / p300.

[0237] In some embodiments, the regulatory molecule is PCAF (P300 / CBP-associated factor).

[0238] In some embodiments, the regulatory molecule is CECR2 (cat eye syndrome chromosomal candidate gene 2).

[0239] In some embodiments, the regulatory molecule is BRPF (bromodomain and PHD finger-containing protein).

[0240] In some embodiments, the regulatory molecule is the ATAD2 or ATAD2B chromatin remodeling protein.

[0241] In some embodiments, the regulatory molecule is BAZ2 (bromodomain adjacent to zinc finger domain).

[0242] In some embodiments, the regulatory molecule is TAF1 (TBP-associated factor).

[0243] In some embodiments, the regulatory molecule is TRIM24 (tripartite motif-containing protein 24).

[0244] In some embodiments, the regulatory molecule is BRD7 / 9.

[0245] In some embodiments, the regulatory molecule is BPTF (bromodomain PHD finger transcription factor).

[0246] In some embodiments, the regulatory molecule is SMARCA2 / 4.

[0247] In some embodiments, the regulatory molecule is PBRM1.

[0248] In some embodiments, the regulatory molecule regulates histone rearrangement.

[0249] In some embodiments, the regulatory molecule regulates histone glycosylation, phosphorylation, alkylation or acylation.

[0250] In some embodiments, the regulatory molecule is a transcription factor.

[0251] In some embodiments, the regulatory molecule is an RNA polymerase.

[0252] In some embodiments, the regulatory molecule is a moiety that regulates the activity of RNA polymerase.

[0253] In some embodiments, the recruitment moiety binds to the regulatory molecule without inhibiting the activity of the regulatory molecule. In some embodiments, the recruitment moiety binds to the regulatory molecule and inhibits the activity of the regulatory molecule. In some embodiments, the recruitment moiety binds to the regulatory molecule and increases the activity of the regulatory molecule.

[0254] In some embodiments, the recruitment moiety binds to the active site of the regulatory molecule. In certain embodiments, the recruitment moiety binds to the regulatory site of the regulatory molecule.

[0255] In some embodiments, the second terminus comprises a structure of formula (2-A) or a pharmaceutically acceptable salt thereof:

[0256]

[0257] Wherein:

[0258] Ring A is an optionally substituted aryl or an optionally substituted 5- to 6-membered heteroaryl;

[0259] Ring B is absent or is an optionally substituted 6-membered monocyclic aryl or heteroaryl;

[0260] D is C or N;

[0261] E is O or N;

[0262] Y A is -NH- or -O-;

[0263] R 5 is hydrogen or a C1-C6 alkyl;

[0264] R 6 is selected from hydrogen, an optionally substituted C1-C6 alkyl, an optionally substituted C1-C6 haloalkyl, and an optionally substituted C1-C6 hydroxyalkyl;

[0265] R 7 is selected from hydrogen, halogen, -NO2, -CN, an optionally substituted aryl, an optionally substituted C1-C 20 alkyl, an optionally substituted C1-C 20 heteroalkyl, an optionally substituted C1-C6 haloalkyl, and an optionally substituted C1-C6 hydroxyalkyl;

[0266] Or R 7 is -NR 7A R 7B wherein R 7A and R 7B are each independently hydrogen, an optionally substituted C1-C 20 alkyl or an optionally substituted C1-C 20 heteroalkyl; and

[0267] x1 is an integer from 1 to 6.

[0268] In some embodiments, D is N and E is N. In some embodiments, D is C and E is O.

[0269] In some embodiments, the second terminus comprises a structure of formula (2-B) or a pharmaceutically acceptable salt thereof:

[0270]

[0271] Wherein:

[0272] Ring A is an optionally substituted aryl or an optionally substituted 5- to 6-membered heteroaryl;

[0273] Ring B is absent or is an optionally substituted 6-membered monocyclic aryl or heteroaryl;

[0274] Y A is -NH- or -O-;

[0275] R 5 is hydrogen or a C1-C6 alkyl group;

[0276] R 6 is selected from hydrogen, an optionally substituted C1-C6 alkyl group, an optionally substituted C1-C6 haloalkyl group, and an optionally substituted C1-C6 hydroxyalkyl group;

[0277] R 7 is selected from hydrogen, halogen, -NO2, -CN, an optionally substituted aryl group, an optionally substituted C1-C 20 alkyl group, any

[0278] optionally substituted C1-C 20 heteroalkyl group, an optionally substituted C1-C6 haloalkyl group, and an optionally substituted C1-C6 hydroxyalkyl group;

[0279] or R 7 is -NR 7A R 7B wherein R 7A and R 7B are each independently hydrogen, an optionally substituted C1-C 20 alkyl group, or an optionally substituted C1-C 20 heteroalkyl group; and

[0280] x1 is an integer from 1 to 6.

[0281] In some embodiments, ring A is an optionally substituted aromatic ring. In some embodiments, ring A is an optionally substituted phenyl group. In some embodiments, ring A is an optionally substituted 5-membered heteroaryl group. In some embodiments, ring A is an optionally substituted oxazolyl group, an optionally substituted furyl group, or an optionally substituted phenylthio group.

[0282] In some embodiments, the second terminus comprises a structure of formula (2-C) or a pharmaceutically acceptable salt thereof:

[0283]

[0284] wherein:

[0285] R 8 and R 9 are each independently selected from hydrogen, an optionally substituted C1-C6 alkyl group, an optionally substituted C1-C6 haloalkane

[0286] yl group, or an optionally substituted C1-C6 hydroxyalkyl group.

[0287] In some embodiments, R 8 and R 9Each independently selected from optionally substituted C1-C6 alkyl, C1-C6 haloalkyl or C1-C6 hydroxyalkyl. In some embodiments, R 8 and R 9 Each independently selected from optionally substituted C1-C6 alkyl. In some embodiments, R 8 and R 9 Each independently is methyl, ethyl or propyl. In some embodiments, R 8 and R 9 Each independently is methyl. In some embodiments, R 8 and R 9 Each independently is ethyl. In some embodiments, R 8 and R 9 Each independently is propyl.

[0288] In some embodiments, the second terminus comprises a structure of formula (2-D) or a pharmaceutically acceptable salt thereof:

[0289]

[0290] Wherein:

[0291] R 10 Selected from hydrogen, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 haloalkyl or optionally substituted C1-C6 hydroxyalkyl.

[0292] Alkyl.

[0293] In some embodiments, R 5 Is C1-C6 alkyl. In some embodiments, R 5 Is methyl or ethyl. In some embodiments, R 5 Is methyl. In some embodiments, R 5 Is ethyl. In some embodiments, R 5 Is hydrogen.

[0294] In some embodiments, R 7 Selected from hydrogen, halogen, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 haloalkyl or optionally substituted C1-C6 hydroxyalkyl. In some embodiments, R 7 Is halogen. In some embodiments, R 7 Is Br, Cl or F. In some embodiments, R 7 Is Cl. In some embodiments, R 7 Is F. In some embodiments, R 7 Is Br.

[0295] In some embodiments, R 7 Is -NR 7AR 7B , wherein R 7A and R 7B are each independently hydrogen or an optionally substituted C1-C6 alkyl group.

[0296] In some embodiments, R 10 is selected from an optionally substituted C1-C6 alkyl group, an optionally substituted C1-C6 haloalkyl group, or an optionally substituted C1-C6 hydroxyalkyl group. In some embodiments, R 10 is selected from an optionally substituted C1-C6 alkyl group. In some embodiments, R 10 is methyl, ethyl, or propyl. In some embodiments, R 10 is methyl. In some embodiments, R 10 is an optionally substituted C 1-6 hydroxyalkyl group. In some embodiments, R 10 is -OMe.

[0297] In some embodiments, R 6 is selected from an optionally substituted C1-C6 alkyl group, an optionally substituted C1-C6 haloalkyl group, or an optionally substituted C1-C6 hydroxyalkyl group. In some embodiments, R 6 is an optionally substituted C1-C6 alkyl group. In some embodiments, R 6 is methyl, ethyl, or propyl. In some embodiments, R 6 is methyl. In some embodiments, R 6 is ethyl. In some embodiments, R 6 is propyl. In some embodiments, R 6 is hydrogen.

[0298] In some embodiments, Y A is -NH-. In some embodiments, Y A is -O-. In some embodiments, Y A is NH and x1 is 1.

[0299] In some embodiments, x1 is an integer from 1 to 5, 1 to 4, 1 to 3, or 1 to 2. In some embodiments, x1 is 1. In some embodiments, x1 is 2.

[0300] In some embodiments, ring B is an optionally substituted 6-membered monocyclic aryl or heteroaryl group, wherein each aryl or heteroaryl group is optionally substituted by an alkyl group, an amino group, a halogen, a hydroxyl group, a hydroxyalkyl group, or a PEG. In some embodiments, ring B is a phenyl group. In some embodiments, ring B is a 6-membered monocyclic heteroaryl group. In some embodiments, ring B is pyridine or pyrimidine. In some embodiments, ring B is absent.

[0301] In some embodiments, the second terminus comprises a structure having formula (2-E), (2-F), or (2-G) or a pharmaceutically acceptable salt thereof:

[0302]

[0303] In some embodiments, the second terminus comprises a structure having formula (3-A) or a pharmaceutically acceptable salt thereof:

[0304]

[0305] wherein:

[0306] Y B is -CH2NH-, -CH2O-, -NH-, or -O-;

[0307] R 11A and R 11B are each independently hydrogen or optionally substituted C1-C6 alkyl;

[0308] R 12 is hydrogen, halogen, -OH, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 haloalkyl, or optionally substituted C1-C6 hydroxyalkyl;

[0309] R 14 and R 15 are each independently hydrogen, halogen, -CN, -NO2, optionally substituted -C1-C6 alkyl, optionally substituted C1-C6 haloalkyl, or optionally substituted C1-C6 hydroxyalkyl;

[0310] or R 14 is -NR A R B ;

[0311] R 16 is optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, C1-C6 hydroxyalkyl, -SO2R A or -NHSO2R A ;

[0312] R YA is hydrogen, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 haloalkyl, or optionally substituted 5- to 6-membered monocyclic aryl or heteroaryl;

[0313] Each R A and R Bindependently is hydrogen, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C1-C6 haloalkyl, optionally substituted C1-C6 hydroxyalkyl, optionally substituted C3-C6 cycloalkyl, optionally substituted 4- to 6-membered heteroalkyl or optionally substituted 4- to 6-membered heteroalkyl; and

[0314] y1 is 1-3;

[0315] wherein the attachment to the linker is at R 14 or R YA therein.

[0316] In some embodiments, the second terminus comprises a structure of formula (3-B) or a pharmaceutically acceptable salt thereof:

[0317]

[0318] wherein:

[0319] ring C is absent, or is an optionally substituted 5- to 6-membered monocyclic aryl or heteroaryl or 4- to 8-membered heterocycle;

[0320] Y B is -NH-, -CH2NH-, -CH2O- or -O-;

[0321] R 11A and R 11B are each independently hydrogen or optionally substituted C1-C6 alkyl;

[0322] R 12 is hydrogen, optionally substituted C1-C6 alkyl, C(O)R A or C(O)NR A R B ; wherein each R A and R B is independently hydrogen, optionally substituted C1-C6 alkyl or optionally substituted C1-C6 heteroalkyl;

[0323] R 13 is hydrogen, substituted aryl, substituted heteroaryl or substituted diphenyl ether; and

[0324] y2 is an integer from 0 to 2.

[0325] In some embodiments, y2 is 0. In some embodiments, y2 is 1. In some embodiments, y2 is 2.

[0326] In some embodiments, R 13 is substituted aryl or substituted heteroaryl. In some embodiments, R 13 is hydrogen.

[0327] In some embodiments, R 13 is a substituted diphenyl ether.

[0328] In some embodiments, R 13 is wherein

[0329] R 14 and R 15 are each independently hydrogen, halogen, -CN, -NO2, optionally substituted -C1-C6 alkyl, optionally substituted C1-C6 haloalkyl or optionally substituted C1-C6 hydroxyalkyl;

[0330] or R 14 is -NR A R B ;

[0331] R 16 is a substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C2-C6 alkenyl,

[0332] optionally substituted C2-C6 alkynyl, optionally substituted C1-C6 hydroxyalkyl, -SO2R A or -NHSO2R A ;

[0333] Each R A and R B is independently hydrogen, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C1-C6 haloalkyl, optionally substituted C1-C6 hydroxyalkyl, optionally substituted C3-C6 cycloalkyl, optionally substituted 4- to 6-membered heteroalkyl or optionally substituted 5- to 6-membered heteroaryl; and

[0334] y1 is 1 - 3.

[0335] In some embodiments, the second terminus comprises a structure of formula (3-C) or a pharmaceutically acceptable salt thereof:

[0336]

[0337] wherein:

[0338] Ring C is absent, or is an optionally substituted 5- to 6-membered monocyclic aryl or heteroaryl or 4- to 8-membered heterocycle;

[0339] Y B is -CH2NH-, -CH2O-, -NH- or -O-;

[0340] R11A and R 11B each independently is hydrogen or an optionally substituted C1-C6 alkyl group;

[0341] R 12 is hydrogen, halogen, -OH, an optionally substituted C1-C6 alkyl group, an optionally substituted C1-C6 haloalkyl group or an optionally substituted C1-C6 hydroxyalkyl group;

[0342] R 14 and R 15 each independently is hydrogen, halogen, -CN, -NO2, an optionally substituted -C1-C6 alkyl group, an optionally substituted C1-C6 haloalkyl group or an optionally substituted C1-C6 hydroxyalkyl group;

[0343] or R 14 is -NR A R B ;

[0344] R 16 is a substituted C1-C6 alkyl group, an optionally substituted C1-C6 heteroalkyl group, an optionally substituted C2-C6 alkenyl group,

[0345] an optionally substituted C2-C6 alkynyl group, an optionally substituted C1-C6 hydroxyalkyl group, -SO2R A or -NHSO2R A ;

[0346] each R A and R B is independently hydrogen, an optionally substituted C1-C6 alkyl group, an optionally substituted C1-C6 heteroalkyl group, an optionally substituted C1-C6 haloalkyl group, an optionally substituted C1-C6 hydroxyalkyl group, an optionally substituted C3-C6 cycloalkyl group, an optionally substituted 4- to 6-membered heteroalkyl group or an optionally substituted 5- to 6-membered heteroaryl group; and

[0347] y1 is an integer from 1 to 3.

[0348] In some embodiments, Y B is -NH-. In some embodiments, Y B is -CH2NH-. In some embodiments, Y B is -CH2O-. In some embodiments, Y B is -O-.

[0349] In some embodiments, ring C is an optionally substituted 5- or 6-membered monocyclic aryl or heteroaryl, wherein each aryl or heteroaryl is optionally substituted with alkyl, amino, halogen, hydroxy, hydroxyalkyl or PEG. In some embodiments, ring C is phenyl. In some embodiments, ring C is a 6-membered heteroaryl. In some embodiments, ring C is pyridine, pyrazine or triazine. In some embodiments, ring C is pyridine. In some embodiments, ring C is pyrazine. In some embodiments, ring C is triazine. In some embodiments, ring C is a 5-membered heteroaryl. In some embodiments, ring C is pyrazole. In some embodiments, ring C is triazole, pyrrole, imidazole, oxazole, oxadiazole, thiazole or thiadiazole. In some embodiments, ring C is triazole. In some embodiments, ring C is imidazole or pyrrole. In some embodiments, oxazole or oxadiazole. In some embodiments, ring C is thiazole or thiadiazole. In some embodiments, ring C is absent.

[0350] In some embodiments, the second terminus comprises a structure of formula (3-D) or a pharmaceutically acceptable salt thereof:

[0351]

[0352] Wherein:

[0353] R 11A and R 11B are each independently hydrogen or an optionally substituted C1-C6 alkyl;

[0354] R 12 is hydrogen, halogen, -OH, an optionally substituted C1-C6 alkyl, an optionally substituted C1-C6 haloalkyl, an optionally substituted C1-C6 hydroxyalkyl, C(O)R A or C(O)NR A R B ;

[0355] Each R 15 is independently hydrogen, halogen, -CN, -NO2, an optionally substituted C1-C6 alkyl, an optionally substituted C1-C6 haloalkyl or an optionally substituted C1-C6 hydroxyalkyl;

[0356] R 16 is an optionally substituted C1-C6 alkyl, an optionally substituted C1-C6 heteroalkyl, optionally

[0357] substituted C2-C6 alkenyl, an optionally substituted C2-C6 alkynyl, an optionally substituted C 1-6 hydroxyalkyl, -SO2R A or -NHSO2R A ;

[0358] Each RA and R B is hydrogen, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally

[0359] substituted C1-C6 haloalkyl, optionally substituted C1-C6 hydroxyalkyl, optionally substituted C3-C6 cycloalkyl, optionally substituted 4- to 6-membered heteroalkyl or optionally substituted 5- to 6-membered heteroaryl; and

[0360] y1 is an integer from 1 to 3.

[0361] In some embodiments, R 11A and R 11B are each independently optionally substituted C1-C6 alkyl. In some embodiments, R 11A and R 11B are each independently methyl, ethyl, propyl or tert-butyl. In some embodiments, R 11A and R 11B are each independently methyl. In some embodiments, R 11A and R 11B are each independently hydrogen.

[0362] In some embodiments, R 11A is C1-C6 alkyl optionally substituted with haloalkyl or phosphorous hydroxide. In some embodiments, R 11A is C1-C6 alkyl substituted with -OP(O)(OH)2. In some embodiments, R 11A is unsubstituted C1-C6 alkyl. In some embodiments, R 11A is methyl, ethyl or tert-butyl. In some embodiments, R 11A is methyl. In some embodiments, R 11A is hydrogen.

[0363] In some embodiments, R 12 is optionally substituted C1-C6 alkyl. In some embodiments, R 12 is hydrogen. In some embodiments, R 12 is C(O)R A or C(O)NR A R B . In some embodiments, R 12 is C(O)NR A R B , wherein R A and R B are each independently hydrogen or optionally substituted C1-C6 alkyl.

[0364] In some embodiments, R 14 and R15 Each independently is hydrogen, -CN or -NO2. In some embodiments, R 14 and R 15 Each independently is a halogen or an optionally substituted C1-C6 alkyl group. In some embodiments, R 14 and R 15 Each independently is Br, Cl, F, methyl or ethyl. In some embodiments, R 14 and R 15 Each independently is F or methyl.

[0365] In some embodiments, R 16 is an optionally substituted C1-C6 alkyl group, an optionally substituted C1-C6 heteroalkyl group, an optionally substituted C2-C6 alkenyl group, an optionally substituted C2-C6 alkynyl group or an optionally substituted C1-C6 hydroxyalkyl group, wherein each group is optionally substituted by an amide group, an alkyl group, an alkynyl group, an azide group, an amino group, a halogen, a haloalkyl group, a hydroxyl group, a nitro group, oxo(=O), phosphoric acid hydroxide or PEG. In some embodiments, R 16 is an optionally substituted C1-C6 alkyl group, an optionally substituted C1-C6 heteroalkyl group or an optionally substituted C1-C6 hydroxyalkyl group. In some embodiments, R 16 is a C1-C6 alkyl group or a C1-C6 heteroalkyl group, wherein each group is optionally substituted by -CN, -NH2, -N3, -OH, CF3 or -OP(O)(OH)2. In some embodiments, R 16 is -SO2R A wherein R A is a C1-C6 alkyl group. In some embodiments, R 16 is -SO2Et. In some embodiments, R 16 is -SO2Me. In some embodiments, R 16 is -NHSO2R A wherein R A is a C1-C6 alkyl group. In some embodiments, R 16 is -NHSO2Et. In some embodiments, R 16 is -NHSO2Me.

[0366] In some embodiments, y1 is 1. In some embodiments, y1 is 2. In some embodiments, y1 is 3.

[0367] In some embodiments, the second terminus comprises a structure of formula (3-E) or (3-F) or a pharmaceutically acceptable salt thereof:

[0368]

[0369] In some embodiments, the second terminus comprises a structure of formula (3-G) or (3-H) or a pharmaceutically acceptable salt thereof:

[0370]

[0371] In some embodiments, the second terminus comprises a structure of formula (4-A) or a pharmaceutically acceptable salt thereof:

[0372]

[0373] Wherein:

[0374] Ring D is absent, or is phenyl or a 5- to 6-membered heteroaryl;

[0375] X 9 and X 10 are each independently C or N, wherein one of X 9 or X 10 is N;

[0376] L 2 is absent, or is optionally substituted alkylene, -O-, or -NR D -, wherein R D is hydrogen or optionally substituted C1-C3 alkyl;

[0377] R 18 is optionally substituted 5- to 6-membered heteroaryl;

[0378] R 19 is optionally substituted C3-C8 cycloalkyl or optionally substituted 4- to 7-membered heteroaryl;

[0379] Each R 20 is independently hydrogen, halogen, -CN, -NO2, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 haloalkyl, or optionally substituted C1-C6 hydroxyalkyl;

[0380] x3 is an integer from 1 to 3; and

[0381] y4 is an integer from 1 to 4;

[0382] Wherein the attachment to the linker is at one of R 19 or R 20 .

[0383] In some embodiments, the attachment to the linker is at R 19 . In some embodiments, the attachment to the linker is at one of R 20 .

[0384] In some embodiments, the second terminus comprises a structure of formula (4-B) or a pharmaceutically acceptable salt thereof:

[0385]

[0386] Wherein:

[0387] Ring D is absent, or is optionally substituted phenyl or optionally substituted 5- to 6-membered heteroaryl;

[0388] X 9 and X 10 are each independently C or N, wherein one of X 9 or X 10 is N;

[0389] L 2 is absent, or is optionally substituted alkylene, -O-, or -NR D -, wherein R D is hydrogen or optionally substituted C1-C3 alkyl;

[0390] R 18 is optionally substituted 5- to 6-membered heteroaryl;

[0391] R 19 is optionally substituted C3-C8 cycloalkyl or optionally substituted 4- to 7-membered heteroaryl; and

[0392] x3 is an integer from 1 to 3.

[0393] In some embodiments, X 9 is N; and X 10 is C. In some embodiments, X 9 is C; and X 10 is N.

[0394] In some embodiments, the second terminus comprises a structure of formula (4-C) or a pharmaceutically acceptable salt thereof:

[0395]

[0396] Wherein:

[0397] Ring D is absent, or is optionally substituted phenyl or optionally substituted 5- to 6-membered heteroaryl;

[0398] L 2 is absent, or is optionally substituted alkylene, -O-, or -NR D -, wherein R D is hydrogen or optionally substituted

[0399] C1-C3 alkyl;

[0400] R 18 is an optionally substituted 5- to 6-membered heteroaryl;

[0401] R 19 is an optionally substituted C3-C8 cycloalkyl or an optionally substituted 4- to 7-membered heteroaryl; and

[0402] x3 is an integer from 1 to 3.

[0403] In some embodiments, ring D is an optionally substituted monocyclic 6-membered aryl or 5- to 6-membered heteroaryl. In some embodiments, ring D is an optionally substituted monocyclic 6-membered aryl. In some embodiments, ring D is an optionally substituted phenyl.

[0404] In some embodiments, R 19 is an optionally substituted C3-C8 cycloalkyl. In some embodiments, R 19 is an optionally substituted 4- to 7-membered heteroaryl.

[0405] In some embodiments, the second terminus comprises a structure of formula (4-D) or a pharmaceutically acceptable salt thereof:

[0406]

[0407] wherein:

[0408] L 2 is an optionally substituted alkylene, -O-, or -NR D -, wherein R D is hydrogen or an optionally substituted C1-C3 alkyl;

[0409] R 18 is an optionally substituted 5- to 6-membered heteroaryl;

[0410] R 20 is hydrogen, halogen, -CN, -NO2, an optionally substituted C1-C6 alkyl, an optionally substituted C1-C6 haloalkyl, or an optionally substituted C1-C6 hydroxyalkyl;

[0411] x3 is an integer from 1 to 3; and

[0412] y4 is an integer from 1 to 4.

[0413] In some embodiments, L 2 is an optionally substituted alkylene. In some embodiments, L 2 is a C2-C4 alkylene optionally substituted by one or more C1-C3 alkyls. In some embodiments, L 2 is absent. In some embodiments, L2 is -NR D -. In some embodiments, L 2 is -NH-.

[0414] In some embodiments, R 18 is an optionally substituted 5 - membered heteroaryl. In some embodiments, R 18 is an optionally substituted oxazole, oxadiazole, thiazole, thiadiazole, pyrrole or pyrazole. In some embodiments, R 18 is an optionally substituted oxazole.

[0415] In some embodiments, R 20 is halogen, -CN, -NO2, an optionally substituted C1 - C6 alkyl, an optionally substituted C1 - C6 haloalkyl or an optionally substituted C1 - C6 hydroxyalkyl.

[0416] In some embodiments, x3 is 1. In some embodiments, x3 is 2. In some embodiments, x3 is 3.

[0417] In some embodiments, y4 is 1 or 2. In some embodiments, y4 is 1. In some embodiments, y4 is 2. In some embodiments, y4 is 3. In some embodiments, y4 is 4.

[0418] In some embodiments, the second terminal comprises a structure having formula (4 - E), (4 - F) or (4 - G) or a pharmaceutically acceptable salt thereof:

[0419]

[0420] In some embodiments, the second terminal comprises a structure having formula (5 - A) or a pharmaceutically acceptable salt thereof:

[0421]

[0422] Wherein:

[0423] Ring E is absent, or is an optionally substituted phenyl or an optionally substituted 5 - to 6 - membered heteroaryl;

[0424] X 11 is CH or N;

[0425] L 3 is -NR E - or -CR E R E -, where each R E is independently hydrogen or an optionally substituted C1 - C3 alkyl;

[0426] R 21is a C1-C6 alkyl or a C3-C6 cycloalkyl; and

[0427] R 22 is hydrogen, CN, NO2, an optionally substituted C1-C6 alkyl, an optionally substituted C1-C6 haloalkyl or an optionally substituted C1-C6 hydroxyalkyl.

[0428]

[0429] In some embodiments, ring E is absent. In some embodiments, ring E is an optionally substituted phenyl. In some embodiments, ring E is an optionally substituted 5- to 6-membered heteroaryl. In some embodiments, ring E is a 5-membered heteroaryl. In some embodiments, ring E is a 6-membered heteroaryl.

[0430] In some embodiments, X 11 is CH and L 3 is -NR E -. In some embodiments, X 11 is N and L 3 is -CR E R E -.

[0431] In some embodiments, R 21 is a C1-C6 alkyl. In some embodiments, R 21 is methyl.

[0432] In some embodiments, R 22 is a halogen, an optionally substituted C1-C6 alkyl, an optionally substituted C1-C6 haloalkyl or an optionally substituted C1-C6 hydroxyalkyl. In some embodiments, R 22 is CN, F, Cl, Br or methyl.

[0433] In some embodiments, the second terminus comprises a structure of formula (5-B) or a pharmaceutically acceptable salt thereof:

[0434]

[0435] In some embodiments, the second terminus comprises a structure of formula (6-A) or a pharmaceutically acceptable salt thereof:

[0436]

[0437] Wherein:

[0438] Ring G is an optionally substituted C3-C6 cycloalkyl or an optionally substituted 4- to 6-membered heterocycloalkyl;

[0439] L 6 is -O-(optionally substituted alkylene);​

[0440] R 28 is an optionally substituted 5- to 6-membered heteroaryl;

[0441] R 29 is an optionally substituted C1-C6 aryl(C6-C 10 aryl) or an optionally substituted C1-C6 alkyl(6- to 10-membered heteroaryl); and

[0442] R 30 is an optionally substituted C1-C6 alkyl, an optionally substituted C1-C6 haloalkyl or an optionally substituted C1-C6 hydroxyalkyl.

[0443] In some embodiments, the second terminus comprises a structure of formula (7-A) or a pharmaceutically acceptable salt thereof:

[0444]

[0445] Wherein:

[0446] A 3 is -O-, -NH- or -CH2-;

[0447] Z 2 is CH or N;

[0448] W is O or S;

[0449] Each R 31 is independently hydrogen, halogen, -OH, -CN, -NO2, -NH2, an optionally substituted C1-C 10 alkyl, an optionally

[0450] substituted C1-C 10 haloalkyl, an optionally substituted C1-C 10 hydroxyalkyl, an optionally substituted C2-C 10 alkenyl, an optionally substituted C2-C 10 alkynyl, an optionally substituted C3-C8 cycloalkyl or an optionally substituted 3- to 8-membered heterocycloalkyl;

[0451] Or two Rs 31 together with the atoms to which they are attached form an optionally substituted C5-C8 cycloalkyl or an optionally substituted 5- to 8-membered heterocycloalkyl;

[0452] R 32 is hydrogen or an optionally substituted C1-C 10 alkyl;

[0453] R 33is hydrogen, a halogen, -OH, -CN, -NO2, -NH2, an optionally substituted C1-C 10 alkyl, an optionally substituted C1-C 10 haloalkyl or an optionally substituted C1-C 10 hydroxyalkyl; and

[0454] q6 is from 0 to 4.

[0455] In some embodiments, the second terminus comprises a structure of formula (7-B) or a pharmaceutically acceptable salt thereof:

[0456]

[0457] wherein:

[0458] Ring F is an optionally substituted 5- to 6-membered heteroaryl;

[0459] A 3 is -O-, -NH- or -CH2-;

[0460] Z 3 is CH or N;

[0461] W is O or S;

[0462] Each R 31 is hydrogen, a halogen, -OH, -CN, -NO2, -NH2, an optionally substituted C1-C 10 alkyl, an optionally

[0463] substituted C1-C 10 haloalkyl, an optionally substituted C1-C 10 hydroxyalkyl, an optionally substituted C2-C 10 alkenyl, an optionally substituted C2-C 10 alkynyl, an optionally substituted C3-C8 cycloalkyl or an optionally substituted 3- to 8-membered heterocycloalkyl;

[0464] Or two R 31 together with the atoms to which they are attached form an optionally substituted C5-C8 cycloalkyl or an optionally substituted 5- to 8-membered heterocycloalkyl;

[0465] R 32 is hydrogen or an optionally substituted C1-C 10 alkyl; and

[0466] q6 is from 1 to 4.

[0467] In some embodiments, A 3 is -O-. In some embodiments, A 3 is -NH-. In some embodiments, A3 is -CH2-.

[0468] In some embodiments, Z 2 is CH. In some embodiments, Z 2 is N.

[0469] In some embodiments, Z 3 is CH. In some embodiments, Z 3 is N.

[0470] In some embodiments, W is O. In some embodiments, W is S.

[0471] In some embodiments, ring F is an optionally substituted 5-membered heteroaryl. In some embodiments, ring F is an optionally substituted 6-membered heteroaryl.

[0472] In some embodiments, each R 31 is independently an optionally substituted C1-C 10 alkyl, an optionally substituted C1-C 10 haloalkyl or an optionally substituted C1-C 10 hydroxyalkyl. In some embodiments, each R 31 is independently an optionally substituted C3-C8 cycloalkyl or an optionally substituted 3- to 8-membered heterocycloalkyl. In some embodiments, each R 31 is independently hydrogen, halogen, -OH, -CN, -NO2 or -NH2. In some embodiments, each R 31 is independently hydrogen.

[0473] In some embodiments, R 32 is an optionally substituted C1-C 10 alkyl. In some embodiments, R 32 is methyl. In some embodiments, R 32 is hydrogen.

[0474] In some embodiments, R 33 is hydrogen, halogen, -OH, -CN, -NO2 or -NH2. In some embodiments, R 33 is an optionally substituted C1-C 10 alkyl, an optionally substituted C1-C 10 haloalkyl or an optionally substituted C1-C 10 hydroxyalkyl.

[0475] In some embodiments, the second terminus comprises a structure of formula (7-C) or a pharmaceutically acceptable salt thereof:

[0476]

[0477] In some embodiments, the second terminus comprises a structure of formula (7-D) or a pharmaceutically acceptable salt thereof:

[0478]

[0479] In some embodiments, the second terminus comprises a structure of formula (8-A) or a pharmaceutically acceptable salt thereof:

[0480]

[0481] Wherein:

[0482] Ring H is an optionally substituted phenyl or an optionally substituted 6-membered heteroaryl;

[0483] Or ring H is

[0484] Z A Absent, or is an optionally substituted phenylformamide;

[0485] X 12 Is CH or N;

[0486] R 34 Is an optionally substituted phenyl or an optionally substituted 6-membered heteroaryl;

[0487] R 34A Is hydrogen, halogen or an optionally substituted C1-C3 alkyl;

[0488] R 35 Independently is halogen, an optionally substituted C1-C 10 Alkyl or an optionally substituted 5- to 6-membered heteroaryl; and

[0489] Wherein the attachment to the linker is at R 35 , Z A Or ring H.

[0490] In some embodiments, ring H is an optionally substituted phenyl. In some embodiments, ring H is an optionally substituted 6-membered heteroaryl. In some embodiments, ring H is

[0491] In some embodiments, Z A Is absent. In some embodiments, Z A Is an optionally substituted phenylformamide. In some embodiments, Z A Is -C(O)NH-phenyl.

[0492] In some embodiments, X 12 Is CH. In some embodiments, X 12 Is N.

[0493] In some embodiments, R 34 is an optionally substituted phenyl. In some embodiments, R 34 is an optionally substituted 6-membered heteroaryl.

[0494] In some embodiments, R 34A is hydrogen or a halogen. In some embodiments, R 34A is an optionally substituted C1-C3 alkyl. In some embodiments, R 34A is methyl.

[0495] In some embodiments, the attachment of formula (8-A) to the linker is at R 35 . In some embodiments, the attachment of formula (8-A) to the linker is at Z A . In some embodiments, the attachment of formula (8-A) to the linker is at ring H.

[0496] In some embodiments, the second terminus comprises a structure having formula (8-B) or (8-C) or a pharmaceutically acceptable salt thereof:

[0497]

[0498] In some embodiments, the second terminus comprises a structure having formula (8-D) or a pharmaceutically acceptable salt thereof:

[0499]

[0500] In some embodiments, the second terminus comprises a structure having formula (9-A) or a pharmaceutically acceptable salt thereof:

[0501]

[0502] In some embodiments, the second terminus comprises a structure having formula (10-A) or (10-B) or a pharmaceutically acceptable salt thereof:

[0503]

[0504] In some embodiments, the second terminus comprises a structure having formula (11-A) or a pharmaceutically acceptable salt thereof:

[0505]

[0506] In some embodiments, the second terminus comprises a structure having formula (12-A) or a pharmaceutically acceptable salt thereof:

[0507]

[0508] Wherein:

[0509] A 4 is -CR 40 R 40 - or -NR 40 -, where each R 40 is independently hydrogen or optionally substituted C1-C 10 alkyl;

[0510] R 36 is optionally substituted 5- to 6-membered heteroaryl;

[0511] Each R 37 is independently hydrogen, halogen, C1-C6 alkyl or C1-C6 haloalkyl;

[0512] R 38 is optionally substituted C1-C 10 alkyl, optionally substituted C3-C8 cycloalkyl or optionally substituted 3- to 8-membered heterocycloalkyl;

[0513] R 39 is hydrogen, halogen, -OH, -CN, -NO2, -NH2, oxo(=O), =S, C1-C 10 haloalkyl or C1-C 10 hydroxyalkyl;

[0514] p 11 is 1 to 4;

[0515] q1 and q2 are each independently 0 to 2; and

[0516] where the linker is attached to formula (12-A) at R 38 or R 40 therein.

[0517] In some embodiments, R 36 is optionally substituted 5-membered heteroaryl. In some embodiments, R 36 is optionally substituted oxazole, oxadiazole, thiazole, thiadiazole, pyrrole or pyrazole. In some embodiments, R 36 is optionally substituted oxazole.

[0518] In some embodiments, each R 37 is independently halogen, C1-C6 alkyl or C1-C6 haloalkyl. In some embodiments, each R 37 is independently halogen.

[0519] In some embodiments, R 38 is optionally substituted C1-C 10 alkyl. In some embodiments, R 38is an optionally substituted C3-C8 cycloalkyl or an optionally substituted 3- to 8-membered heterocycloalkyl. In some embodiments, R 38 is a 3- to 8-membered heterocycloalkyl.

[0520] In some embodiments, R 39 is hydrogen, halogen, -OH, -CN, -NO2, -NH2, C1-C 10 haloalkyl or C1-C 10 hydroxyalkyl. In some embodiments, R 39 is oxo or =S. In some embodiments, R 39 is oxo. In some embodiments, R 39 is =S.

[0521] In some embodiments, A 4 is -NR 40 In some embodiments, A 4 is -NH. In some embodiments, A 6 is -NCH3. In some embodiments, A 4 is -CR 40 R 40 In some embodiments, A 4 is -CH2-.

[0522] In some embodiments, each R 40 is independently an optionally substituted C1-C 10 alkyl. In some embodiments, each R 40 is independently hydrogen.

[0523] In some embodiments, p 11 is 3 or 4. In some embodiments, p 11 is 2. In some embodiments, p 11 is 1.

[0524] In some embodiments, q1 is 1 and q2 is 1. In some embodiments, q1 is 2 and q2 is 0.

[0525] In some embodiments, the linker is attached to formula (12-A) through R 38 In some embodiments, the linker is attached to formula (12-A) through R 40 In some embodiments, the second terminus comprises a structure having formula (12-B) or formula (12-C) or a pharmaceutically acceptable salt thereof:

[0526]

[0527]

[0528] ​In some embodiments, the second terminus comprises a structure of formula (12-D) or formula (12-E) or a pharmaceutically acceptable salt thereof:

[0529]

[0530] In some embodiments, the second terminus comprises a structure of formula (13-A) or a pharmaceutically acceptable salt thereof:

[0531]

[0532] Wherein:

[0533] Ring J is absent or is an optionally substituted 5- to 6-membered heteroaryl;

[0534] R 41 is an optionally substituted C1-C6 alkyl, an optionally substituted C3-C8 cycloalkyl, -C(O)R 41a , -C(O)- or -C(O)NR 41a R 41b , wherein

[0535] R 41a and R 41b are each independently an optionally substituted C1-C 10 alkyl or an optionally substituted C3-C8 cycloalkyl;

[0536] R 42 is an optionally substituted C1-C 10 alkyl, an optionally substituted C1-C 10 haloalkyl, an optionally substituted C3-C8 cycloalkyl or an optionally substituted 3- to 8-membered heterocyclic alkyl;

[0537] R 43 is hydrogen or an optionally substituted C1-C 10 alkyl;

[0538] Each R 44 is independently hydrogen, halogen, -OH, -CN, -NO2, -NH2, an optionally substituted C1-C 10 alkyl, an optionally substituted C1-C 10 haloalkyl, C1-C 10 hydroxyalkyl, an optionally substituted C2-C 10 alkenyl, an optionally substituted C2-C 10 alkynyl, an optionally substituted C3-C8 cycloalkyl or an optionally substituted 3- to 8-membered heterocyclic alkyl; or

[0539] R 43 and R 44One, together with the atom to which it is attached, forms an optionally substituted 5- to 8-membered heterocycloalkyl;

[0540] p 12 is from 1 to 4;

[0541] q3 is 0 or 1; and

[0542] wherein formula (13-A) is attached to the linker at ring J or R 41 at this point.

[0543] In some embodiments, R 41 is optionally substituted C1-C6 alkyl or optionally substituted C3-C8 cycloalkyl. In some embodiments, R 41 is -C(O)R 41a . In some embodiments, R 41 is -C(O)CH3 or -C(O)CH2CH3. In some embodiments, R 41 is -C(O)-NR 41a R 41b .

[0544] In some embodiments, R 41a is optionally substituted C1-C 10 alkyl. In some embodiments, R 41a is optionally substituted C3-C8 cycloalkyl.

[0545] In some embodiments, R 41b is optionally substituted C1-C 10 alkyl. In some embodiments, R 41b is optionally substituted C3-C8 cycloalkyl.

[0546] In some embodiments, R 42 is optionally substituted C1-C 10 alkyl or optionally substituted C1-C 10 haloalkyl. In some embodiments, R 42 is optionally substituted C3-C8 cycloalkyl or optionally substituted 3- to 8-membered heterocycloalkyl. In some embodiments, R 42 is optionally substituted 3- to 8-membered heterocycloalkyl ring.

[0547] In some embodiments, R 43 is optionally substituted C1-C 10 alkyl. In some embodiments, R 43 is hydrogen.

[0548] In some embodiments, each R 44Independently is halogen, -OH, -CN, -NO2, -NH2, optionally substituted C1-C 10 alkyl, optionally substituted C1-C 10 haloalkyl, C1-C 10 hydroxyalkyl, optionally substituted C2-C 10 alkenyl, optionally substituted C2-C 10 alkynyl, optionally substituted C3-C8-cycloalkyl or optionally substituted 3- to 8-membered heterocycle. In some embodiments, each R 44 is independently halogen or C1-C 10 haloalkyl.

[0549] In some embodiments, one of R 43 and R 44 forms, together with the atom to which they are attached, an optionally substituted 5- to 8-membered heterocycloalkyl. In some embodiments, one of R 43 and R 44 forms, together with the atom to which they are attached, a 5-, 6-, 7- or 8-membered heterocycloalkyl.

[0550] In some embodiments, p 12 is 3 or 4. In some embodiments, p 12 is 2. In some embodiments, p 12 is 1.

[0551] In some embodiments, q3 is 1. In some embodiments, q3 is 0.

[0552] In some embodiments, ring J is an optionally substituted 5-membered heteroaryl. In some embodiments, ring J is absent.

[0553] In some embodiments, formula (13-A) is attached to the linker at ring J. In some embodiments, formula (13-A) is attached to the linker at R 41 .

[0554] In some embodiments, the second terminus comprises a structure of formula (13-B) or a pharmaceutically acceptable salt thereof:

[0555]

[0556]

[0557] In some embodiments, the second terminus comprises a structure of formula (13-C1) or (13-C2) or a pharmaceutically acceptable salt thereof:

[0558]

[0559] In some embodiments, the second terminus comprises a structure of formula (13-D1) or (13-D2) or a pharmaceutically acceptable salt thereof:

[0560]

[0561] In some embodiments, the second terminus comprises a structure of formula (14-A) or a pharmaceutically acceptable salt thereof:

[0562]

[0563] Wherein:

[0564] Ring K is a 5- to 6-membered heteroalkyl;

[0565] A 5 is absent, or is CH2, -NH-, or -O-;

[0566] L 4 is an alkylene or heteroalkylene;

[0567] Each R 45 is independently halogen, -OH, -CN, -NO2, -NH2, optionally substituted C1-C 10 alkyl, optionally

[0568] substituted C1-C 10 haloalkyl, C1-C 10 hydroxyalkyl, optionally substituted C2-C 10 alkenyl, optionally substituted C2-C 10 alkynyl, optionally substituted C3-C8 cycloalkyl or optionally substituted 3- to 8-membered hetero

[0569] cycloalkyl;

[0570] Each R 46 is independently hydrogen, halogen, -OH, -CN, -NO2, -NH2, optionally substituted C1-C 10 alkyl, any

[0571] optionally substituted C1-C 10 haloalkyl, optionally substituted C1-C 10 heteroalkyl, optionally substituted C1-C 10 hydroxyalkyl, optionally substituted C2-C 10 alkenyl, optionally substituted C2-C 10 alkynyl, optionally substituted C3-C8 cycloalkyl or optionally substituted 3- to 8-membered heteroalkyl;

[0572] R 47 is optionally substituted C1-C10 alkyl, -C(O)R 47a or -C(O)-NR 47a R 47b , wherein

[0573] R 47a and R 47b are each independently optionally substituted C1-C 10 alkyl or optionally substituted C3-C8 cycloalkyl;

[0574] q4 is 2 to 3; or

[0575] q5 is 0 to 2; and

[0576] wherein formula (14-A) is attached to the linker through ring K or through one of the Rs 45 one.

[0577] In some embodiments, A 5 is absent. In some embodiments, A 5 is -NH- or -O-. In some embodiments, A 5 is -NH-. In some embodiments, A 5 is -O-.

[0578] In some embodiments, L 4 is alkylene. In some embodiments, L 4 is C1-C5 alkylene. In some embodiments, L 4 is heteroalkylene. In some embodiments, L 4 is C1-C4 heteroalkylene-. In some embodiments, L 4 is -O-CH2- or -O-CH2CH2-.

[0579] In some embodiments, each R 45 is independently halogen, -OH, -CN, -NO2, -NH2, optionally substituted C1-C 10 alkyl, optionally substituted C1-C 10 haloalkyl, C1-C 10 hydroxyalkyl, optionally substituted C2-C 10 alkenyl or optionally substituted C2-C 10 alkynyl. In some embodiments, each R 45 is independently optionally substituted C1-C 10 alkyl or optionally substituted C1-C 10 hydroxyalkyl. In some embodiments, each R 45 is independently C1-C 10 hydroxyalkyl. In some embodiments, each R 45Independently, it is -OCH3 or -OCH2CH3.

[0580] In some embodiments, each R 46 Independently, it is hydrogen, halogen, -OH, -CN, -NO2, -NH2, optionally substituted C1-C 10 alkyl, optionally substituted C1-C 10 haloalkyl, optionally substituted C1-C 10 heteroalkyl or optionally substituted C1-C 10 hydroxyalkyl. In some embodiments, each R 46 Independently, it is -OCH3. In some embodiments, each R 46 Independently, it is hydrogen.

[0581] In some embodiments, R 47 is optionally substituted C1-C 10 alkyl. In some embodiments, R 47 is -C(O)R 47a In some embodiments, R 47 is -C(O)CH3 or -C(O)CH2CH3. In some embodiments, -C(O)-NR 47a R 47b .

[0582] In some embodiments, R 47a is optionally substituted C1-C 10 alkyl. In some embodiments, R 47a is optionally substituted C3-C8 cycloalkyl.

[0583] In some embodiments, R 47b is optionally substituted C1-C 10 alkyl. In some embodiments, R 47b is optionally substituted C3-C8 cycloalkyl.

[0584] In some embodiments, ring K is a 6-membered heteroalkyl.

[0585] In some embodiments, q4 is 3. In some embodiments, q4 is 2.

[0586] In some embodiments, q5 is 2. In some embodiments, q5 is 1. In some embodiments, q5 is 0.

[0587] In some embodiments, formula (14-A) is connected to the linker through ring K. In some embodiments, formula (14-A) is connected to the linker through one of R 45 .

[0588] In some embodiments, the second terminus comprises a structure having formula (14-B) or (14-C) or a pharmaceutically acceptable salt thereof:

[0589]

[0590] In some embodiments, the second terminus comprises a structure having formula (15-A) or a pharmaceutically acceptable salt thereof:

[0591]

[0592] wherein:

[0593] Ring L is aryl or heteroaryl;

[0594] Each R 48 is hydrogen, halogen, -OH, -CN, -NO2, -NH2, C1-C 10 alkyl, C1-C 10 haloalkyl or C1-C 10 hydroxyalkyl;

[0595] R 49 and R 50 are each independently hydrogen, optionally substituted C1-C 10 alkyl, optionally substituted C1-C 10 haloalkyl, optionally substituted C2-C 10 alkenyl or optionally substituted C2-C 10 alkynyl;

[0596] R 51 is hydrogen, halo, -OH, -CN, -NO2, -NH2, optionally substituted C1-C 10 alkyl, optionally substituted C1-C 10 haloalkyl, optionally substituted C1-C 10 hydroxyalkyl, optionally substituted C2-C 10 alkenyl or optionally substituted C2-C 10 alkynyl;

[0597] R 52 is hydrogen, halogen, -OH, -CN, -NO2, -NH2, optionally substituted C1-C 10 alkyl, optionally substituted C1-C 10 haloalkyl or optionally substituted C1-C 10 hydroxyalkyl; and

[0598] p7 is from 1 to 4.

[0599] In some embodiments, ring L is an aryl group. In some embodiments, the aryl group is a phenyl group. In some embodiments, ring L is a heteroaryl group. In some embodiments, ring L is a bicyclic heteroaryl group containing 1, 2, or 3 heteroatoms selected from N and O.

[0600] In some embodiments, the second terminus comprises a structure of formula (15-B) or a pharmaceutically acceptable salt thereof:

[0601]

[0602] Wherein:

[0603] Each R 48 is hydrogen, halogen, -OH, -CN, -NO2, -NH2, C1-C 10 alkyl, C1-C 10 haloalkyl, or C1-C 10 hydroxyalkyl;

[0604] R 49 and R 50 are each independently hydrogen, optionally substituted C1-C 10 alkyl, optionally substituted C1-C 10 haloalkyl, optionally substituted C2-C 10 alkenyl, or optionally substituted C2-C 10 alkynyl;

[0605] R 51 is hydrogen, halo, -OH, -CN, -NO2, -NH2, optionally substituted C1-C 10 alkyl, optionally substituted C1-C 10 haloalkyl, optionally substituted C1-C 10 hydroxyalkyl, optionally substituted C2-C 10 alkenyl, or optionally substituted C2-C 10 alkynyl; and

[0606] p7 is from 1 to 4.

[0607] In some embodiments, the second terminus comprises a structure of formula (15-C) or a pharmaceutically acceptable salt thereof:

[0608]

[0609] Wherein:

[0610] X is CR 48 or N;

[0611] Each R 48 is hydrogen, halogen, -OH, -CN, -NO2, -NH2, C1-C10 alkyl, C1-C 10 haloalkyl or C1-C 10 hydroxyalkyl;

[0612] R 49 and R 50 are each independently hydrogen, optionally substituted C1-C 10 alkyl, optionally substituted C1-C 10 haloalkyl, optionally substituted C2-C 10 alkenyl or optionally substituted C2-C 10 alkynyl;

[0613] R 51 is hydrogen, halo, -OH, -CN, -NO2, -NH2, optionally substituted C1-C 10 alkyl, optionally substituted C1-C 10 haloalkyl, optionally substituted C1-C 10 hydroxyalkyl, optionally substituted C2-C 10 alkenyl or optionally substituted C2-C 10 alkynyl;

[0614] R 53 is hydrogen or optionally substituted C1-C 10 alkyl; and

[0615] p7 is from 1 to 3.

[0616] In some embodiments, each R 48 is hydrogen, halogen, -OH, -CN, -NO2, -NH2, C1-C 10 alkyl, C1-C 10 haloalkyl or optionally substituted C1-C 10 hydroxyalkyl.

[0617] In some embodiments, R 49 is hydrogen, optionally substituted C1-C 10 alkyl, optionally substituted C1-C 10 haloalkyl, optionally substituted C2-C 10 alkenyl or optionally substituted C2-C 10 alkynyl. In some embodiments, R 49 is optionally substituted C1-C 10 alkyl. In some embodiments, R 49 is methyl, ethyl, isopropyl or tert-butyl. In some embodiments, R 49 is hydrogen.

[0618] In some embodiments, R 50is hydrogen, optionally substituted C1-C 10 alkyl, optionally substituted C1-C 10 haloalkyl, optionally substituted C2-C 10 alkenyl or optionally substituted C2-C 10 alkynyl. In some embodiments, R 50 is optionally substituted C1-C 10 alkyl or optionally substituted C2-C 10 alkenyl. In some embodiments, R 50 is hydrogen.

[0619] In some embodiments, R 51 is hydrogen, halogen, -OH, -CN, -NO2, -NH2, optionally substituted C1-C 10 alkyl, optionally substituted C1-C 10 haloalkyl, optionally substituted C1-C 10 hydroxyalkyl, optionally substituted C2-C 10 alkenyl or optionally substituted C2-C 10 alkynyl.

[0620] In some embodiments, R 52 is hydrogen, halogen, -OH, -CN, -NO2, -NH2, optionally substituted C1-C 10 alkyl, optionally substituted C1-C 10 haloalkyl or optionally substituted C1-C 10 hydroxyalkyl. In some embodiments, R 52 is hydrogen.

[0621] In some embodiments, R 53 is hydrogen or optionally substituted C1-C 10 alkyl. In some embodiments, R 53 is optionally substituted C1-C 10 alkyl. In some embodiments, R 53 is methyl, ethyl, isopropyl or tert-butyl. In some embodiments, R 53 is hydrogen.

[0622] In some embodiments, p7 is 4. In some embodiments, p7 is 3. In some embodiments, p7 is 2. In some embodiments, p7 is 1.

[0623] In some embodiments, the second terminus comprises a structure having formula (15-D1), (15-D2) or (15-D3) or a pharmaceutically acceptable salt thereof:

[0624]

[0625] In some embodiments, the second terminus comprises a structure having formula (15-E1), (15-E2), or (15-E3) or a pharmaceutically acceptable salt thereof:

[0626]

[0627] In some embodiments, the second terminus comprises a structure having formula (16-A) or a pharmaceutically acceptable salt thereof:

[0628]

[0629] Wherein:

[0630] B 5 is -O-, -NH-, or S;

[0631] B 6 is N or CH;

[0632] R 54 is optionally substituted aryl or optionally substituted heteroaryl;

[0633] Each R 55 is independently hydrogen, -OH, -CN, -NO2, -NH2, optionally substituted C1-C 10 alkyl, optionally substituted C1-C 10 haloalkyl, optionally substituted C1-C 10 heteroalkyl, or optionally substituted C1-C 10 hydroxyalkyl;

[0634] R 56 is hydrogen, optionally substituted C1-C 10 alkyl, optionally substituted C1-C 10 haloalkyl, or optionally substituted C1-C 10 hydroxyalkyl;

[0635] R 57 is halogen, -OH, -CN, -NO2, -NH2, optionally substituted C1-C 10 alkyl, optionally substituted C1-C 10 haloalkyl, or optionally substituted C1-C 10 hydroxyalkyl;

[0636] p9 is 1 to 3; and

[0637] q7 is 0 to 2.

[0638] In some embodiments, B 5 is -O- or -S-. In some embodiments, B5 is -O-. In some embodiments, B 5 is -S-.

[0639] In some embodiments, B 6 is N. In some embodiments, B 6 is CH.

[0640] In some embodiments, R 54 is optionally substituted aryl. In some embodiments, R 54 is optionally substituted with one or more halogens, -CN, -NH2, -OH, C1-C 10 alkyl, C1-C 10 haloalkyl or C1-C 10 hydroxyalkyl substituted phenyl.

[0641] In some embodiments, each R 55 is independently halogen, -OH, -CN, -NH2, optionally substituted C1-C 10 alkyl, optionally substituted C1-C 10 haloalkyl, optionally substituted C1-C 10 heteroalkyl or optionally substituted C1-C 10 hydroxyalkyl.

[0642] In some embodiments, R 56 is optionally substituted C1-C 10 alkyl, optionally substituted C1-C 10 haloalkyl or optionally substituted C1-C 10 hydroxyalkyl. In some embodiments, R 56 is optionally substituted C1-C 10 alkyl.

[0643] In some embodiments, R 57 is halogen, -OH, -CN, -NO2, -NH2 or optionally substituted C1-C 10 alkyl.

[0644] In some embodiments, p9 is 3. In some embodiments, p9 is 2. In some embodiments, p9 is 1.

[0645] In some embodiments, q7 is 2. In some embodiments, q7 is 1. In some embodiments, q7 is 0.

[0646] In some embodiments, the second terminus comprises a structure of formula (16-B) or a pharmaceutically acceptable salt thereof:

[0647]

[0648] In some embodiments, the second terminus comprises a structure of formula (17-A) or a pharmaceutically acceptable salt thereof:

[0649]

[0650] Wherein:

[0651] Ring M is an optionally substituted aryl or an optionally substituted heteroaryl;

[0652] Ring N is absent or is a 4- to 8-membered heterocycloalkyl;

[0653] A 6 is -O-, -NH-, or -CH2-;

[0654] Each R 58 is independently halogen, -OH, -CN, -NO2, -NH2, C1-C 10 alkyl, C1-C 10 haloalkyl, or C1-C 10 hydroxyalkyl;

[0655] R 59 is hydrogen, -OH, -NH2, C1-C 10 alkyl, C1-C 10 haloalkyl, C1-C 10 hydroxyalkyl, or -NH-C1-C 10 alkyl;

[0656] R 60 is hydrogen or an optionally substituted C1-C 10 alkyl; and

[0657] p 10 is 1 to 4; and

[0658] wherein formula (17-A) is attached to the linker through R 59 connected to the linker.

[0659] In some embodiments, Ring M is an aryl optionally substituted with one or more halogen, CN, NH2, OH, C1-C 10 alkyl, C1-C 10 haloalkyl, or C1-C 10 hydroxyalkyl. In some embodiments, Ring M is phenyl. In some embodiments, Ring M is a 6-membered heteroaryl optionally substituted with one or more halogen, CN, NH2, OH, C1-C 10 alkyl, C1-C 10 haloalkyl, or C1-C 10 hydroxyalkyl. In some embodiments, Ring M is an optionally substituted pyridine.

[0660] In some embodiments, ring N is a 4- to 8-membered heterocycloalkyl. In some embodiments, ring N is a 4-membered heterocycloalkyl. In some embodiments, ring N is a 5-membered heterocycloalkyl. In some embodiments, ring N is a 6-membered heterocycloalkyl. In some embodiments, ring N is absent.

[0661] In some embodiments, A 6 is -O- or -NH-. In some embodiments, A 6 is -CH2-.

[0662] In some embodiments, each R 58 is independently -OH, -NH2, C1-C 10 alkyl, C1-C 10 haloalkyl or C1-C 10 hydroxyalkyl. In some embodiments, each R 58 is independently C1-C 10 alkyl or C1-C 10 hydroxyalkyl. In some embodiments, each R 58 is independently C1-C 10 hydroxyalkyl.

[0663] In some embodiments, R 59 is -OH, -NH2, C1-C 10 hydroxyalkyl or -NH-C1-C 10 alkyl. In some embodiments, R 59 is hydrogen.

[0664] In some embodiments, R 60 is optionally substituted C1-C 10 alkyl. In some embodiments, R 60 is methyl. In some embodiments, R 60 is hydrogen.

[0665] In some embodiments, p 10 is 3 or 4. In some embodiments, p 10 is 2. In some embodiments, p 10 is 1.

[0666] In some embodiments, the second terminus comprises a structure of formula (17-B) or a pharmaceutically acceptable salt thereof:

[0667]

[0668] In some embodiments, the second terminus comprises a structure of formula (17-C) or a pharmaceutically acceptable salt thereof:

[0669]

[0670] In some embodiments, the second terminus is selected from the group consisting of:

[0671]

[0672]

[0673] or a pharmaceutically acceptable salt thereof.

[0674] oligomeric linker

[0675] The length of the oligomeric linker depends on the type of regulatory protein and also on the target gene. In some embodiments, the linker has a length of less than about 50 angstroms. In some embodiments, the linker has a length of about 20 to 30 angstroms. In some embodiments, the oligomeric linker comprises between 5 and 50 backbone atoms.

[0676] In some embodiments, the oligomeric linker comprises a polymer having 2 to 50 spacer moieties, wherein

[0677] each spacer moiety is independently selected from the group consisting of: -((CR 1b R 1b ) x -O) y -, -((CR 1b R 1b ) x -NR 1a ) y -, -((CR 1b R 1b ) x -CH=CH-(CR 1b R 1b ) x -O) y -, optionally substituted C1-C 12 alkyl, optionally substituted C2-C 10 alkenyl, optionally substituted C2-C 10 alkynyl, optionally substituted C6-C 10 arylene, optionally substituted C3-C7 cycloalkylene, optionally substituted 5- to 10-membered heteroarylene, optionally substituted 4- to 10-membered heterocycloalkylene, amino acid residue, -O-, -C(O)NR 1a -, -NR 1a C(O)-, -C(O)-, -NR 1a -, -C(O)O-, -S-, -S(O)-, -S(O)2-, -S(O)2NR 1a -, -NR 1aS(O)2- and -P(O)OH- and any combination thereof; wherein

[0678] each x is independently 2 to 4;

[0679] each y is independently 1 to 10;

[0680] each R 1a is independently hydrogen or optionally substituted C1-C6 alkyl; and

[0681] each R 1b is independently selected from hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted alkoxy, optionally substituted amino, carboxyl, carboxyl ester, acyl, acyloxy, acylamino, aminoacyl, optionally substituted alkylamide, sulfonyl, optionally substituted thioalkoxy, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted cycloalkyl and optionally substituted heterocyclic group.

[0682] In some embodiments, the oligomeric linker comprises a polymer having 2 to 50 spacer moieties, wherein each spacer moiety is independently selected from the group consisting of optionally substituted C1-C 12 alkyl, -((CH2) x -O) y -, -((CH2) x -NH) y -, -O-, -C(O)NH-, -NH- and combinations thereof.

[0683] In various aspects, a composition comprising a double-stranded DNA binding moiety for treating genetic diseases is suitable for topical administration. In some embodiments, the composition is suitable for administration to the human eye. In some embodiments, the composition further comprises an excipient. In some embodiments, the excipient includes an isotonicity regulator (such as sodium chloride), a buffer, a stabilizer, an antioxidant, a thickening agent, a solubility enhancer, a penetration enhancer and / or a preservative. In some embodiments, the pH of the composition is about 5 to about 8. In some embodiments, the viscosity of the composition at about 20 °C is about 1 to about 50,000 cps.

[0684] In all aspects, administration of a composition comprising a double-stranded DNA binding moiety for treating genetic diseases to the eye results in minimal systemic exposure. In some embodiments, topical ocular administration (e.g., ophthalmic administration) of the composition results in penetration of the double-stranded DNA binding moiety into the internal cells and tissues of the eye. In some embodiments, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 95% or at least about 99% of the double-stranded DNA binding moiety administered to the eye can penetrate the cells and tissues inside the eye.

[0685] Aqueous solution dose-dose uniformity

[0686] Typical ophthalmic aqueous solutions, emulsions or suspensions are packaged in eye drop bottles and administered as drops. For example, a single administration (i.e., single dose) of an ophthalmic aqueous solution, emulsion or suspension comprises instilling one drop, two drops, three drops or more drops into the patient's eye. In some embodiments, one dose of the ophthalmic aqueous solution, emulsion or suspension described herein is one drop of the aqueous solution, emulsion or suspension composition from an eye drop bottle.

[0687] In some cases, the ophthalmic aqueous compositions described herein can provide dose-to-dose uniform concentrations. In some cases, there is no significant change in the drug content from one dose to another in the dose-to-dose uniform concentration. In some cases, the dose-to-dose uniform concentration provides a consistent drug content from one dose to another.

[0688] In some embodiments, the dose-to-dose ophthalmic pharmaceutical concentration variation of the composition is less than 50%. In some embodiments, the dose-to-dose ophthalmic pharmaceutical concentration variation of the composition is less than 40%. In some embodiments, the dose-to-dose ophthalmic pharmaceutical concentration variation of the composition is less than 30%. In some embodiments, the dose-to-dose ophthalmic pharmaceutical concentration variation of the composition is less than 20%. In some embodiments, the dose-to-dose ophthalmic pharmaceutical concentration variation of the composition is less than 10%. In some embodiments, the dose-to-dose ophthalmic pharmaceutical concentration variation of the composition is less than 5%.

[0689] In some embodiments, the dose-to-dose ophthalmic pharmaceutical concentration variation is based on 10 consecutive doses. In some embodiments, the dose-to-dose ophthalmic pharmaceutical concentration variation is based on 8 consecutive doses. In some embodiments, the dose-to-dose ophthalmic pharmaceutical concentration variation is based on 5 consecutive doses. In some embodiments, the dose-to-dose ophthalmic pharmaceutical concentration variation is based on 3 consecutive doses. In some embodiments, the dose-to-dose ophthalmic pharmaceutical concentration variation is based on 2 consecutive doses.

[0690] A non-sedimenting composition should not require shaking to disperse the drug evenly. "No-shake" compositions may be advantageous compared to compositions that require shaking, simply because the shaking behavior of the patient is a major source of variability in the drug dosage administered. It has been reported that, despite clear labeling instructions to shake, patients generally do not or forget to shake compositions that require shaking before administering the dose. On the other hand, even for those patients who do shake the product, it is generally not possible to determine whether the shaking intensity and / or duration is sufficient to homogenize the product. In some embodiments, the ophthalmic gel compositions and ophthalmic ointment compositions described herein are "no-shake" formulations that maintain the dose-to-dose uniformity described herein. In some embodiments, the ophthalmic gel compositions and ophthalmic ointment compositions described herein require shaking to maintain the dose-to-dose uniformity described herein.

[0691] To evaluate dose-to-dose uniformity, the dropper bottles or tubes containing the ophthalmic aqueous composition, ophthalmic gel composition, or ophthalmic ointment composition were placed upright until 12 hours before the test began. To simulate the recommended doses of these products, a predetermined number of drops or strips were dispensed from each commercially available bottle or tube at predetermined time intervals, or until there was no product left in the bottle or tube. All drops and strips were dispensed into tared glass vials, capped, and stored at room temperature until analysis. The concentration of the double-stranded DNA-binding moiety in the expressed droplets was determined using a reverse-phase HPLC method.

[0692] Aqueous solution viscosity

[0693] In some embodiments, the composition has a Brookfield RVDV viscosity of about 1 to about 50,000 centipoise (cps) at about 20 °C and a shear rate of 1 s -1 . In some embodiments, the composition has a Brookfield RVDV viscosity of about 100 to about 40,000 cps at about 20 °C and a shear rate of 1 s -1 . In some embodiments, the composition has a Brookfield RVDV viscosity of about 500 to about 30,000 cps at about 20 °C and a shear rate of 1 s -1 . In some embodiments, the composition has a Brookfield RVDV viscosity of about 1000 to about 20,000 cps at about 20 °C and a shear rate of 1 s -1 . In some embodiments, the composition has a Brookfield RVDV viscosity of about 2000 to about 10,000 cps at about 20 °C and a shear rate of 1 s -1 . In some embodiments, the composition has a Brookfield RVDV viscosity of about 4000 to about 8000 cps at about 20 °C and a shear rate of 1 s -1 .

[0694] In some embodiments, the aqueous composition contains a thickening agent sufficient to provide a viscosity between about 500 and 50,000 cps, between about 750 and 50,000 cps, between about 1000 and 50,000 cps, between about 1000 and 40,000 cps, between about 2000 and 30,000 cps, between about 3000 and 20,000 cps, between about 4000 and 10,000 cps, or between about 5000 and 8000 cps.

[0695] In some embodiments, the aqueous composition comprises a viscosity reducing agent. In some embodiments, the viscosity reducing agent comprises caffeine. In some embodiments, the aqueous composition comprises a suitable concentration of caffeine suitable to achieve the desired viscosity. In some embodiments, the composition comprises about 0.5%, about 1%, about 1.5%, about 2%, about 2.5%, about 3%, about 3.5%, about 4%, about 4.5%, about 5%, or about 10% caffeine.

[0696] In some embodiments, the compositions described herein are low viscosity compositions at body temperature. In some embodiments, the low viscosity composition contains from about 1% to about 10% thickening agent (e.g., a gelling component such as a polyethylene glycol-oxypropylene copolymer). In some embodiments, the low viscosity composition contains from about 2% to about 10% thickening agent (e.g., a gelling component such as a polyethylene glycol-oxypropylene copolymer). In some embodiments, the low viscosity composition contains from about 5% to about 10% thickening agent (e.g., a gelling component such as a polyethylene glycol-oxypropylene copolymer). In some embodiments, the low viscosity composition is substantially free of thickening agent (e.g., a gelling component such as a polyethylene glycol-oxypropylene copolymer). In some embodiments, the low viscosity ophthalmic pharmaceutical composition described herein provides an apparent viscosity of from about 100 cps to about 10,000 cps. In some embodiments, the low viscosity ophthalmic pharmaceutical composition described herein provides an apparent viscosity of from about 500 cps to about 10,000 cps. In some embodiments, the low viscosity ophthalmic pharmaceutical composition described herein provides an apparent viscosity of from about 1000 cps to about 10,000 cps.

[0697] Osmotic pressure

[0698] In some embodiments, the compositions disclosed herein are formulated so as not to disrupt the ionic balance of the eye. In some embodiments, the compositions disclosed herein have the same or substantially the same ionic balance as the eye. In some embodiments, the compositions disclosed herein do not disrupt the ionic balance of the eye.

[0699] As used herein, "actual osmotic pressure / osmolality" or "deliverable osmotic pressure / osmolality" means the osmotic pressure / osmolality of a composition determined by measuring the osmotic pressure / osmolality of the ophthalmic agent and all excipients except gelling and / or thickening agents (such as polyethylene glycol-oxypropylene copolymers, carboxymethylcellulose, etc.). The actual osmotic pressure of the compositions disclosed herein is measured by a suitable method, such as the freezing point depression method described in Viegas et al., Int. J. Pharm., 1998, 160, 157-162. In some cases, the actual osmotic pressure of the compositions disclosed herein is measured by vapor pressure osmometry (e.g., vapor pressure lowering method), which allows the determination of the osmotic pressure of the composition at a higher temperature. In some cases, the vapor pressure lowering method can allow the determination of the osmotic pressure of a composition containing a gelling agent (e.g., a thermoreversible polymer) at a higher temperature, where the gelling agent is in the form of a gel.

[0700] In some embodiments, the osmotic pressure of the target site of action (e.g., the eye) is approximately the same as the osmotic pressure of the compositions described herein. In some embodiments, the deliverable osmotic pressure of the compositions described herein is from about 150 mOsm / L to about 500 mOsm / L, from about 250 mOsm / L to about 500 mOsm / L, from about 250 mOsm / L to about 350 mOsm / L, from about 280 mOsm / L to about 370 mOsm / L, or from about 250 mOsm / L to about 320 mOsm / L.

[0701] The actual osmolality of the compositions disclosed herein is from about 100 mOsm / kg to about 1000 mOsm / kg, from about 200 mOsm / kg to about 800 mOsm / kg, from about 250 mOsm / kg to about 500 mOsm / kg, or from about 250 mOsm / kg to about 320 mOsm / kg or from about 250 mOsm / kg to about 350 mOsm / kg or from about 280 mOsm / kg to about 320 mOsm / kg. In some embodiments, the actual osmotic pressure of the compositions described herein is from about 100 mOsm / L to about 1000 mOsm / L, from about 200 mOsm / L to about 800 mOsm / L, from about 250 mOsm / L to about 500 mOsm / L, from about 250 mOsm / L to about 350 mOsm / L, from about 250 mOsm / L to about 320 mOsm / L, or from about 280 mOsm / L to about 320 mOsm / L.

[0702] In some embodiments, suitable tonicity modifiers include, but are not limited to, any pharmaceutically acceptable sugar, salt, or any combination or mixture thereof, such as, but not limited to, dextrose, glycerin, mannitol, sorbitol, sodium chloride, and other electrolytes. In some cases, the tonicity modifier is selected from sodium chloride, sodium nitrate, sodium sulfate, sodium bisulfate, potassium chloride, calcium chloride, magnesium chloride, zinc chloride, potassium acetate, sodium acetate, sodium bicarbonate, sodium carbonate, sodium thiosulfate, magnesium sulfate, disodium hydrogen phosphate, sodium dihydrogen phosphate, potassium dihydrogen phosphate, dextrose, mannitol, sorbitol, dextrose, sucrose, urea, propylene glycol, glycerin, or combinations thereof.

[0703] In some embodiments, the compositions described herein include an amount of one or more salts sufficient to bring the osmolarity of the composition to an acceptable range. Such salts include salts containing sodium, potassium, or ammonium cations and chloride, citrate, ascorbate, borate, phosphate, bicarbonate, sulfate, thiosulfate, or bisulfite anions; suitable salts include sodium chloride, potassium chloride, sodium thiosulfate, sodium bisulfite, and ammonium sulfate.

[0704] Sterility

[0705] In some embodiments, the composition is sterilized. Embodiments disclosed herein include the means and processes for sterilization of pharmaceutical compositions for human use disclosed herein. The aim is to provide safe pharmaceutical products that are relatively free from microbial contamination. The U.S. Food and Drug Administration provides regulatory guidance in the publication "Guidance for Industry: Sterile Drug Products Produced by Aseptic Processing", available at: http: / / www.fda.gov / cder / guidance / 5882fnl.htm, which is incorporated herein by reference in its entirety.

[0706] As used herein, sterilization means a process for destroying or removing microorganisms present in the product or package. Any suitable method available for sterilization of the object and composition is used. Available methods for microbial inactivation include, but are not limited to, the application of extreme heat, lethal chemicals, or γ-radiation. In some embodiments, the process of preparing an ophthalmic composition includes subjecting the composition to a sterilization method selected from heat sterilization, chemical sterilization, radiation sterilization, or filtration sterilization. The method used depends largely on the nature of the equipment or composition to be sterilized. Chapter 40 of Remington: The Science and Practice of Pharmacy, published by Lippincott, Williams & Wilkins, describes in detail many sterilization methods and is incorporated herein by reference on this subject.

[0707] Filtration sterilization

[0708] Filtration sterilization is a method used to remove but not destroy microorganisms in a solution. Membrane filters are used to filter heat-sensitive solutions. Such filters are thin, strong, homogeneous polymers of mixed cellulose esters (MCE), polyvinylidene fluoride (PVF; also known as PVDF), or polytetrafluoroethylene (PTFE), with pore sizes ranging from 0.1 to 0.22 μm. Different filter membranes can be optionally used to filter solutions with various characteristics. For example, PVF and PTFE membranes are very suitable for filtering organic solvents, while aqueous solutions are filtered through PVF or MCE membranes. Filter devices can be used on many scales, from single-point disposable filters attached to syringes to commercial-scale filters for manufacturing plants. The membrane filters are sterilized by autoclaving or chemical sterilization. The validation of the membrane filtration system follows a standardized protocol (Microbiological Evaluation of Filters for Sterilizing Liquids, Volume 4, Issue 3 Washington, D.C: Health Industry Manufacturers Association, 1981) and involves challenging the membrane filter with a known quantity (about 10 7 / cm 2 ) of extremely small microorganisms, such as Brevundimonas diminuta (ATCC 19146).

[0709] Optionally, a pharmaceutical composition can be sterilized by passing it through a membrane filter. Compositions containing nanoparticles (U.S. Patent No. 6,139,870) or multilamellar vesicles (Richard et al., International Journal of Pharmaceutics (2006), 312(1-2): 144-50) can be sterilized by filtration through a 0.22 μm filter without destroying their tissue structure.

[0710] In some embodiments, the methods disclosed herein include sterilizing a composition (or its components) by filtration sterilization. In an ophthalmic gel composition comprising a thermosetting polymer, a peristaltic pump is used to perform the filtration at a temperature below (e.g., about 5°C) the gel temperature (Tgel) of the composition described herein and a viscosity that allows filtration within a reasonable time (e.g., below 100 cps of the theoretical value).

[0711] Accordingly, the present disclosure provides methods for sterilizing the compositions provided herein to prevent degradation of polymeric components (such as thermosetting agents and / or other tackifiers) and / or ophthalmic agents during the sterilization process. In some embodiments, degradation of an ophthalmic agent (such as a double-stranded DNA binding moiety) is reduced or eliminated by using a buffer component in a specific pH range and a specific ratio of tackifier in the composition. In some embodiments, selection of an appropriate tackifier or thermosetting polymer allows for sterilization of the compositions described herein by filtration. In some embodiments, use of an appropriate thermosetting polymer or other tackifier in combination with a specific pH range of the composition allows for high-temperature sterilization of the described composition with substantially no degradation of the therapeutic agent or polymer excipient. An advantage of the sterilization methods provided herein is that, in certain cases, the composition is terminally sterilized via autoclaving without loss of the ophthalmic agent and / or excipient and / or tackifier during the sterilization process and is substantially free of microorganisms and / or pyrogens.

[0712] Sterilization by radiation

[0713] One advantage of sterilization by radiation is the ability to disinfect a wide variety of product types without thermal degradation or other damage. The radiation commonly used is β-radiation, or alternatively γ-radiation from a 60 Co source. The penetrating ability of γ-radiation allows its use in the sterilization of many product types, including solutions, compositions, and heterogeneous mixtures. The bactericidal action of radiation results from the interaction of γ-radiation with biological macromolecules. This interaction produces charged particle species and free radicals. Subsequent chemical reactions, such as rearrangement and crosslinking processes, lead to the loss of normal function of these biological macromolecules. The compositions described herein are also optionally sterilized using β-radiation.

[0714] Sterilization by heating

[0715] Sterilization can be carried out using a number of methods by applying high heat. One method is to use an autoclave with saturated steam. In this method, saturated steam at a temperature of at least 121 °C is allowed to contact the object to be sterilized. Heat transfer is either directly to the microorganisms on the object to be sterilized or indirectly to the microorganisms by heating the bulk of the aqueous solution to be sterilized. This method is widely practiced because it allows flexibility, safety, and economy during the sterilization process.

[0716] Microorganisms

[0717] In some embodiments, the composition is substantially free of microorganisms. Acceptable bioburden or sterility levels are based on applicable standards that define a therapeutically acceptable composition, including but not limited to United States Pharmacopeia chapter <1111> and below. For example, acceptable sterility (e.g., bioburden) levels include about 10 colony forming units (cfu) per gram of composition, about 50 cfu per gram of composition, about 100 cfu per gram of composition, about 500 cfu per gram of composition, or about 1000 cfu per gram of composition. In some embodiments, acceptable bioburden levels or sterilization of the composition include less than 10 cfu / mL, less than 50 cfu / mL, less than 500 cfu / mL, or less than 1000 cfu / mL of microbial agents. Additionally, acceptable bioburden levels or sterilization include exclusion of designated objectionable microbial agents. By way of example, designated objectionable microbial agents include, but are not limited to, Escherichia coli (E. coli), Salmonella species, Pseudomonas aeruginosa (P. aeruginosa), and / or other specific microbial agents.

[0718] An important component of the sterilization assurance quality control, quality assurance, and validation processes is the method of sterility testing. Sterility testing is performed by two methods (by way of example only). The first is direct inoculation, where a sample of the composition to be tested is added to a growth medium and incubated for up to 21 days. Turbidity of the growth medium indicates contamination. Disadvantages of this method include a smaller sampling size for large amounts of material, which reduces sensitivity, and detection of microbial growth based on visual observation. An alternative method is membrane filtration sterility testing. In this method, a quantity of the product is passed through a small membrane filter paper. The filter paper is then placed in a medium to promote microbial growth. With sampling of the entire large quantity of product, this method has greater sensitivity. The commercially available Millipore Steritest sterility testing system is optionally used to determine by membrane filtration sterility testing. For filtration testing of creams or ointments, the Steritest filtration system number TLHVSL210 is used. For filtration testing of emulsions or viscous products, the Steritest filtration system numbers TLAREM210 or TDAREM210 are used. For filtration testing of prefilled syringes, the Steritest filtration system number TTHASY210 is used. For filtration testing of materials dispensed as aerosols or foams, the Steritest filtration system number TTHVA210 is used. For filtration testing of soluble powders in ampoules or vials, the Steritest filtration system numbers TTHADA210 or TTHADV210 are used.

[0719] Tests for Escherichia coli and Salmonella include using lactose broth incubated at 30 to 35 °C for 24 to 72 hours, incubation in MacConkey and / or EMB agar for 18 to 24 hours, and / or using Rappaport medium. Tests for detecting Pseudomonas aeruginosa include using NAC agar. Chapter <62> of the United States Pharmacopeia further enumerates test procedures for specifying objectionable microorganisms.

[0720] In certain embodiments, the compositions described herein have less than about 60 colony forming units (cfu), less than about 50 colony forming units, less than about 40 colony forming units, or less than about 30 colony forming units of microbial agent per gram of formulation. In certain embodiments, the compositions described herein are formulated to be isotonic with the eye.

[0721] Endotoxin

[0722] Another aspect of the sterilization process is the removal of by-products from killed microorganisms. The process of pyrogen removal removes pyrogens from a sample. Pyrogens are endotoxins or exotoxins that induce an immune response. An example of an endotoxin is the lipopolysaccharide (LPS) molecule found in the cell wall of Gram-negative bacteria. Although there are sterilization procedures such as autoclaving or treatment with ethylene oxide that kill bacteria, LPS residues can induce a pro-inflammatory immune response such as septic shock. Due to the large variation in the molecular size of endotoxins, the presence of endotoxins is expressed as "endotoxin units" (EU). One EU is equivalent to 100 picograms of E. coli LPS. In some cases, humans have responded to as little as 5 EU / kg of body weight. Bioburden (e.g., microbial limit) and / or sterility (e.g., endotoxin level) are expressed in any units recognized in the art. In certain embodiments, the compositions described herein contain a lower endotoxin level (e.g., <4 EU / kg of subject body weight) compared to a conventionally acceptable endotoxin level (e.g., 5 EU / kg of subject body weight). In one embodiment, the composition has less than about 5 EU / kg of subject body weight. In one embodiment, the composition has less than about 4 EU / kg of subject body weight. In one embodiment, the composition has less than about 3 EU / kg of subject body weight. In one embodiment, the composition has less than about 2 EU / kg of subject body weight.

[0723] In one embodiment, the composition has less than about 5 EU / kg of the composition. In other embodiments, the composition has less than about 4 EU / kg of the composition. In further embodiments, the composition has less than about 3 EU / kg of the composition. In one embodiment, the composition has less than about 2 EU / kg of the composition. In other embodiments, the composition has less than about 1 EU / kg of the composition. In further embodiments, the composition has less than about 0.2 EU / kg of the composition. In certain embodiments, the composition described herein contains from about 1 to about 5 EU / mL of the composition. In certain embodiments, the composition described herein contains from about 2 to about 5 EU / mL of the composition, from about 3 to about 5 EU / mL of the composition, or from about 4 to about 5 EU / mL of the composition.

[0724] In certain embodiments, compared to a conventionally acceptable endotoxin level (e.g., 0.5 EU / mL of the composition), the composition described herein contains a lower endotoxin level (e.g., <0.5 EU / mL of the composition). In one embodiment, the composition has less than about 0.5 EU / mL of the composition. In other embodiments, the composition has less than about 0.4 EU / mL of the composition. In further embodiments, the composition has less than about 0.2 EU / mL of the composition.

[0725] Pyrogen testing is performed by several methods (by way of example only). Suitable tests for sterilization include those described in the United States Pharmacopeia (USP) <71> Sterility Tests (23rd Edition, 1995). Both the rabbit pyrogen test and the Limulus amebocyte lysate test are described in USP Chapters <85> and <151> (USP23 / NF 18, Biological Tests, The United States Pharmacopeial Convention, Rockville, Maryland, 1995). Alternative pyrogen assays have been developed based on monocyte activation - cytokine assays. A homogeneous cell line has been developed for quality control applications and demonstrated the ability to detect pyrogenicity in samples passing the rabbit pyrogen test and the Limulus amebocyte lysate test (Taktak et al., J. Pharm. Pharmacol. (1990), 43:578 - 82). In further embodiments, the composition is depyrogenated. In further embodiments, the process of manufacturing the composition includes testing the pyrogenicity of the composition. In certain embodiments, the composition described herein is substantially free of pyrogens.

[0726] Ophthalmic gel composition

[0727] Gels have been defined in various ways. For example, the United States Pharmacopeia defines a gel as a suspension composed of small inorganic particles or a semi-solid system composed of large organic molecules penetrated by a liquid. Gels include single-phase or two-phase systems. Single-phase gels are composed of organic macromolecules uniformly distributed throughout the liquid, i.e., there is no distinct boundary between the dispersed macromolecules and the liquid. Some single-phase gels are prepared from synthetic macromolecules (such as carbomer) or natural gums (such as tragacanth). In some embodiments, single-phase gels are typically aqueous, but gels made from alcohols and oils will also be used. Two-phase gels consist of a network of small discrete particles.

[0728] In some embodiments, gels are also classified as hydrophobic or hydrophilic. In certain embodiments, non-limiting examples of the matrix of hydrophobic gels include liquid paraffin containing polyethylene, or fatty oils gelled with colloidal silica, or aluminum or zinc soaps. In contrast, non-limiting examples of the matrix of hydrophilic gels include water, glycerol, or propylene glycol gelled with a suitable gelling agent (such as tragacanth, starch, cellulose derivatives, carboxyvinyl polymer, and magnesium aluminum silicate). In certain embodiments, the rheology of the compositions disclosed herein is pseudoplastic, plastic, thixotropic, or dilatant.

[0729] In some embodiments, the composition is an ophthalmic gel, and an ophthalmically acceptable carrier therein includes water and at least one thickening agent. In some embodiments, the thickening agent is selected from cellulose-based polymers, polyethylene glycol-polypropylene oxide triblock polymers, dextran-based polymers, polyvinyl alcohol, dextran, polyvinylpyrrolidone, polyalkenyl alcohol, chitosan, collagen, gelatin, hyaluronic acid, or a combination thereof.

[0730] In some embodiments, the composition contains a viscosity reducing agent. In some embodiments, the viscosity reducing agent contains caffeine. In some embodiments, the composition contains an appropriate concentration of caffeine to achieve the desired viscosity. In some embodiments, the composition contains about 0.5%, about 1%, about 1.5%, about 2%, about 2.5%, about 3%, about 3.5%, about 4%, about 4.5%, about 5%, or about 10% of caffeine.

[0731] In some embodiments, the ophthalmic gel composition described herein is semi-solid or in a gel state (e.g., at room temperature) before topical administration. For example, suitable thickening agents for such gels include (by way of example only) gelling agents and suspending agents. In one embodiment, the viscosity enhancing composition does not include a buffer. In other embodiments, the viscosity enhancing composition includes a pharmaceutically acceptable buffer. If necessary, sodium chloride or other tonicity agents are optionally used to adjust the tonicity.

[0732] By way of example only, ophthalmically acceptable viscosifiers include hydroxypropyl methylcellulose, hydroxyethyl cellulose, polyvinylpyrrolidone, carboxymethyl cellulose, polyvinyl alcohol, sodium chondroitin sulfate, or sodium hyaluronate. Other thickeners compatible with the targeted ocular location include, but are not limited to, gum arabic (acacia), agar, magnesium aluminum silicate, sodium alginate, sodium stearate, fucus, bentonite, carbomer, carrageenan, Carbopol, xanthan gum, cellulose, microcrystalline cellulose (MCC), carob gum, chitosan, carboxymethyl chitosan, chondrus, dextrose, furcellaran, gelatin, ghatti gum, guar gum, lithium montmorillonite, lactose, sucrose, maltodextrin, mannitol, sorbitol, honey, corn starch, wheat starch, rice starch, potato starch, gelatin, sterculia gum, xanthan gum, tragacanth gum, ethyl cellulose, ethylhydroxyethyl cellulose, ethylmethyl cellulose, methyl cellulose, hydroxyethyl cellulose, hydroxyethylmethyl cellulose, hydroxypropyl cellulose, poly(hydroxyethyl methacrylate), oxidized gelatin, pectin, polypeptone, polyvinylpyrrolidone, acrylate, methyl vinyl ether / maleic anhydride copolymer (PVM / MA), poly(methoxyethyl methacrylate), poly(methoxyethoxyethyl methacrylate), hydroxypropyl cellulose, hydroxypropyl methylcellulose (HPMC), sodium carboxymethyl cellulose (CMC), silica, polyvinylpyrrolidone (PVP: polyvinylpyrrolidone), (dextrose, maltodextrin, and sucralose) or combinations thereof. In certain embodiments, the thickening excipient is a combination of MCC and CMC. In another embodiment, the thickener is a combination of carboxymethyl chitosan or chitosan and alginate. The combination of chitosan and alginate with the ophthalmic agent disclosed herein functions as a controlled release composition that limits the diffusion of the ophthalmic agent from the composition. Additionally, the combination of carboxymethyl chitosan and alginate optionally helps increase the permeability of the agent in the eye.

[0733] In some embodiments, the present disclosure provides an enhanced viscosity composition comprising an ophthalmic agent at about 0.1 mM to about 100 mM, a pharmaceutically acceptable viscosity agent, and water for injection, wherein the concentration of the viscosity agent in the water is sufficient to provide an enhanced viscosity composition having a final viscosity of about 100 to about 100,000 cps. In certain embodiments, the viscosity of the gel is in the range of about 100 to about 50,000 cps, about 100 cps to about 1,000 cps, about 500 cps to about 1500 cps, about 1000 cps to about 3000 cps, about 2000 cps to about 8,000 cps, about 4,000 cps to about 50,000 cps, about 10,000 cps to about 500,000 cps, or about 15,000 cps to about 1,000,000 cps. In other embodiments, when a more viscous medium is desired, the biocompatible gel comprises at least about 35%, at least about 45%, at least about 55%, at least about 65%, at least about 70%, at least about 75%, or at least about 80% (by weight) of the ophthalmic agent. In high concentration samples, the biocompatible enhanced viscosity composition comprises at least about 25%, at least about 35%, at least about 45%, at least about 55%, at least about 65%, at least about 75%, at least about 85%, at least about 90%, or at least about 95% or more (by weight) of the ophthalmic agent.

[0734] In one embodiment, a pharmaceutically acceptable enhanced viscosity ophthalmically acceptable composition comprises at least one ophthalmic agent and at least one gelling agent. Suitable gelling agents used in the preparation of the gel composition include, but are not limited to, cellulose, cellulose derivatives, cellulose ethers (such as carboxymethyl cellulose, ethyl cellulose, hydroxyethyl cellulose, hydroxymethyl cellulose, hydroxypropyl methyl cellulose, hydroxypropyl cellulose, methyl cellulose), guar gum, xanthan gum, tamarind gum, alginates (such as alginic acid), silicates, starches, tragacanth, carboxyvinyl polymers, carrageenans, paraffin, petrolatum, and any combination or mixture thereof. In some other embodiments, hydroxypropyl methyl cellulose is used as the gelling agent. In certain embodiments, the thickening agent described herein is also used as the gelling agent for the gel composition presented herein.

[0735] In some embodiments, the ophthalmic gel composition described herein is an in-situ gel composition. In some cases, the in-situ gel composition is based on increasing the pre-corneal residence time of the composition, thereby improving ocular bioavailability, corneal adhesion, lysosomal interaction, and ion gelation, improving corneal absorption, thermogelation, or a combination thereof. In some cases, the in-situ gel formulation is activated by pH, temperature, ions, ultraviolet light, or solvent exchange.

[0736] In some cases, the ophthalmic gel composition comprises a double-stranded DNA binding moiety and one or more gelling agents. In some cases, the gelling agents include, but are not limited to, poloxamers (e.g., poloxamer 407), tetronics, ethyl(hydroxyethyl)cellulose, cellulose acetate phthalate (CAP), carbomers (e.g., Carbopol 1342P NF, Carbopol980NF), alginates (e.g., low acetyl gellan gum ), gellan gum, hyaluronic acid, pluronics (e.g., Pluronic F-127), chitosan, polyvinyl alcohol (PVA), polyvinylpyrrolidone (PVP), dextran, hydroxypropyl methylcellulose (HPMC), hydroxyethyl cellulose (HEC), methylcellulose (MC), thioglucan, polymethacrylic acid (PMMA), polyethylene glycol (PEG), pseudo-latex, xyloglucan, or combinations thereof.

[0737] In certain cases, the in-situ gel composition further comprises a penetration enhancer. In some cases, the penetration enhancers include surfactants (e.g., nonionic surfactants), benzalkonium chloride, EDTA, surfactant heteroglycosides, calcium chelators, hydroxypropyl β-cyclodextrin (HPβCD), bile salts, etc.

[0738] In some embodiments, other gel compositions are useful, depending on the particular ophthalmic agent, other agents or excipients / additives used, and are thus considered to be within the scope of the present disclosure. For example, other commercially available glycerol-based gels, glycerol-derived compounds, conjugated or crosslinked gels, matrices, hydrogels and polymers, as well as gelatin and its derivatives, alginates and alginate-based gels, and even various natural and synthetic hydrogels and hydrogel-derived compounds, are expected to be used in the ophthalmic agent compositions described herein. In some embodiments, ophthalmically acceptable gels include, but are not limited to, alginate hydrogels (ConvaTec, Princeton, NJ), HydroactiveGel(ConvaTec), (Johnson&Johnson Medical, Arlington, TX); (V)Acemannan Hydrogel(Carrington Laboratories, Inc., Irving, TX); glycerol gel Hydrogel(Swiss-American Products, Inc., Dallas, TX) and Sterile (Johnson & Johnson). In additional embodiments, the biodegradable biocompatible gel also represents a compound present in the ophthalmologically acceptable compositions disclosed and described herein.

[0739] In some embodiments, the thickening agent is a cellulose-based polymer selected from: cellulose gum, alkylcellulose, hydroxyalkylcellulose, hydroxyalkylalkylcellulose, carboxyalkylcellulose, or combinations thereof. In some embodiments, the thickening agent is hydroxyalkylalkylcellulose. In some embodiments, the thickening agent is hydroxypropylmethylcellulose.

[0740] In certain embodiments, the viscosity-enhancing composition is characterized by a phase change between room temperature and body temperature (including individuals with severe fever, e.g., up to about 42 °C). In some embodiments, the phase change occurs at 1 °C below body temperature, at 2 °C below body temperature, at 3 °C below body temperature, at 4 °C below body temperature, at 6 °C below body temperature, at 8 °C below body temperature, or at 10 °C below body temperature. In some embodiments, the phase change occurs at 15 °C below body temperature, below about 20 °C below body temperature, or below about 25 °C below body temperature. In specific embodiments, the gelling temperature (Tgel) of the compositions described herein is about 20 °C, about 25 °C, or about 30 °C. In certain embodiments, the gelling temperature (Tgel) of the compositions described herein is about 35 °C or about 40 °C. Body temperature in the definition includes the body temperature of a healthy or unhealthy individual, including fever (up to about 42 °C). In some embodiments, the pharmaceutical compositions described herein are liquid at about room temperature and are administered at room temperature or at about room temperature.

[0741] The copolymers polyoxypropylene and polyethylene glycol (e.g., polyethylene glycol-polyoxypropylene triblock copolymer) form a thermosetting gel, which is incorporated into an aqueous solution. These polymers have the ability to change from a liquid state to a gel state at temperatures close to body temperature, thus allowing the application of useful compositions to targeted ocular locations. The phase change from the liquid state to the gel state depends on the polymer concentration and the components in the solution.

[0742] In some embodiments, the amount of the thermosetting polymer in any of the compositions described herein is about 10%, about 15%, about 20%, about 25%, about 30%, about 35% or about 40% of the total amount of the composition. In some embodiments, the amount of the thermosetting polymer in any of the compositions described herein is about 10%, about 11%, about 12%, about 13%, about 14%, about 15%, about 16%, about 17%, about 18%, about 19%, about 20%, about 21%, about 22%, about 23%, about 24% or about 25% of the total amount of the composition. In some embodiments, the amount of the thermosetting polymer (e.g., Poloxamer 407) in any of the compositions described herein is about 7.5% of the total weight of the composition. In some embodiments, the amount of the thermosetting polymer (e.g., Poloxamer 407) in any of the compositions described herein is about 10% of the total weight of the composition. In some embodiments, the amount of the thermosetting polymer (e.g., Poloxamer 407) in any of the compositions described herein is about 11% of the total weight of the composition. In some embodiments, the amount of the thermosetting polymer (e.g., Poloxamer 407) in any of the compositions described herein is about 12% of the total weight of the composition. In some embodiments, the amount of the thermosetting polymer (e.g., Poloxamer 407) in any of the compositions described herein is about 13% of the total weight of the composition. In some embodiments, the amount of the thermosetting polymer (e.g., Poloxamer 407) in any of the compositions described herein is about 14% of the total weight of the composition. In some embodiments, the amount of the thermosetting polymer (e.g., Poloxamer 407) in any of the compositions described herein is about 15% of the total weight of the composition. In some embodiments, the amount of the thermosetting polymer (e.g., Poloxamer 407) in any of the compositions described herein is about 16% of the total weight of the composition. In some embodiments, the amount of the thermosetting polymer (e.g., Poloxamer 407) in any of the compositions described herein is about 17% of the total weight of the composition. In some embodiments, the amount of the thermosetting polymer (e.g., Poloxamer 407) in any of the compositions described herein is about 18% of the total weight of the composition. In some embodiments, the amount of the thermosetting polymer (e.g., Poloxamer 407) in any of the compositions described herein is about 19% of the total weight of the composition. In some embodiments, the amount of the thermosetting polymer (e.g., Poloxamer 407) in any of the compositions described herein is about 20% of the total weight of the composition. In some embodiments, the amount of the thermosetting polymer (e.g., Poloxamer 407) in any of the compositions described herein is about 21% of the total weight of the composition. In some embodiments, the amount of the thermosetting polymer (e.g., Poloxamer 407) in any of the compositions described herein is about 22% of the total weight of the composition.In some embodiments, the amount of the thermosetting polymer (e.g., Poloxamer 407) in any of the compositions described herein is about 23% of the total weight of the composition. In some embodiments, the amount of the thermosetting polymer (e.g., Poloxamer 407) in any of the compositions described herein is about 24% of the total weight of the composition. In some embodiments, the amount of the thermosetting polymer (e.g., Poloxamer 407) in any of the compositions described herein is about 25% of the total weight of the composition. In some embodiments, the amount of the thickening agent (e.g., gelling agent) in any of the compositions described herein is about 1%, about 5%, about 10%, or about 15% of the total weight of the composition. In some embodiments, the amount of the thickening agent (e.g., gelling agent) in any of the compositions described herein is about 0.5%, about 1%, about 1.5%, about 2%, about 2.5%, about 3%, about 3.5%, about 4%, about 4.5%, or about 5% of the total weight of the composition.

[0743] In alternative embodiments, the thermogel is a PEG-PLGA-PEG triblock copolymer (Jeong et al., Nature (1997), 388:860-2; Jeong et al., J. Control. Release (2000), 63:155-63; Jeong et al., Adv. Drug Delivery Rev. (2002), 54:37-51). The polymer exhibits solution-gel behavior at concentrations from about 5% w / w to about 40% w / w. Depending on the desired properties, the molar ratio of lactide / glycolide in the PLGA copolymer ranges from about 1:1 to about 20:1. The resulting copolymer is water-soluble and forms a free-flowing liquid at room temperature, but forms a hydrogel at body temperature. A commercially available PEG-PLGA-PEG triblock copolymer is RESOMERR GP t50106, manufactured by Boehringer Ingelheim. The material consists of a PLGA copolymer, 50:50 poly(DL-lactide-co-glycolide), 10% w / w of PEG, and has a molecular weight of about 6000.

[0744] Additional biodegradable thermoplastic polyesters include (Provided by Atrix Laboratories, Inc.) and / or disclosed in the following documents, such as U.S. Patent Nos. 5,324,519; 4,938,763; 5,702,716; 5,744,153; and 5,990,194; wherein suitable biodegradable thermoplastic polyesters are disclosed as thermoplastic polymers. Examples of suitable biodegradable thermoplastic polyesters include polylactide, polyglycolide, polycaprolactone, their copolymers, their terpolymers, and any combination thereof. In some such embodiments, the suitable biodegradable thermoplastic polyester is polylactide, polyglycolide, their copolymers, their terpolymers, or a combination thereof. In one embodiment, the biodegradable thermoplastic polyester is 50 / 50 poly(DL-lactide-co-glycolide) having carboxyl end groups; present at about 30 wt.% to about 40 wt.% of the composition; and having an average molecular weight of about 23,000 to about 45,000. Alternatively, in another embodiment, the biodegradable thermoplastic polyester is 75 / 25 poly(DL-lactide-co-glycolide) without carboxyl end groups; present at about 40 wt.% to about 50 wt.% of the composition; and having an average molecular weight of about 15,000 to about 24,000. In additional or alternative embodiments, depending on the polymerization method, the end groups of poly(DL-lactide-co-glycolide) are hydroxyl, carboxyl, or ester. The polycondensation of lactic acid or glycolic acid provides a polymer having terminal hydroxyl and carboxyl groups. The ring-opening polymerization of cyclic lactide or glycolide monomers with water, lactic acid, or glycolic acid provides a polymer having the same end groups. However, the ring-opening of cyclic monomers with a monofunctional alcohol (such as methanol, ethanol, or 1-dodecanol) provides a polymer having one hydroxyl group and one ester end group. The ring-opening polymerization of cyclic monomers with a diol (such as 1,6-hexanediol or polyethylene glycol) provides a polymer having only hydroxyl end groups.

[0745] Since the polymer system of the thermosetting gel dissolves more completely at reduced temperatures, a dissolution method including adding a desired amount of the polymer to a certain amount of water is used at reduced temperatures. Generally, after wetting the polymer by shaking, the mixture is capped and placed in a cold room or a thermostatic container (about 0 °C to 10 °C) to dissolve the polymer. The mixture is stirred or shaken to dissolve the thermosetting gel polymer more quickly. Subsequently, ophthalmic agents and various additives, such as buffers, salts, and preservatives, are added and dissolved. In some cases, if the agent is insoluble in water, it can be suspended. The pH is adjusted by adding a suitable buffer.

[0746] Ophthalmic ointment composition

[0747] An ointment is a homogeneous, viscous, semi-solid preparation, most commonly an oleaginous, greasy one (e.g., 80% oil - 20% water), with a relatively high viscosity, intended for external application to the skin or mucous membranes. The water number of an ointment defines the maximum amount of water it contains. They are used as emollients or to apply active ingredients to the skin for protective, therapeutic, or prophylactic purposes and require a certain degree of absorption. Ointments are applied locally to various body surfaces. These include the skin and mucous membranes of the eye (ophthalmic ointments), vulva, anus, and nose.

[0748] The vehicle of an ointment is called the ointment base. The choice of base depends on the clinical indication of the ointment. Different types of ointment bases are hydrocarbon bases such as paraffin wax, soft paraffin, microcrystalline wax, and ceresin wax; absorbent bases such as wool fat, beeswax; water-soluble bases such as polyethylene glycol 200, 300, 400; emulsifying bases such as emulsifying wax, cetrimide; vegetable oils such as olive oil, coconut oil, sesame oil, almond oil, and peanut oil.

[0749] Ointments are formulated using hydrophobic, hydrophilic, or water-emulsifying bases to provide preparations that are immiscible, miscible, or emulsifiable with skin secretions. In some embodiments, they are also derived from hydrocarbon (fatty), absorbent, water-removable, or water-soluble bases. The active pharmaceutical agent is dispersed in the base and then dispensed after the drug has penetrated to the target site (e.g., membrane, skin, etc.).

[0750] The present disclosure recognizes that it is sometimes difficult to incorporate low concentrations of a drug into an ointment and have dose - dose uniformity sufficient to effectively treat a condition or disease. In some embodiments, poly(ethylene glycol), polyethoxylated castor oil ( EL), an alcohol having 12 to 20 carbon atoms, or a mixture of two or more of the foregoing components are effective excipients for dispersing and / or dissolving an effective amount of an ophthalmic drug. The resulting ointment is well tolerated by skin and ocular tissues.

[0751] The present disclosure further recognizes that when the composition is topically applied to the ocular surface, particularly the sclera of the patient, an ophthalmic drug (such as a double-stranded DNA-binding moiety) incorporated in the ointment composition described herein targets the choroid and / or retina. In some embodiments, an ophthalmic ointment composition comprises an ophthalmic drug, an ointment base, and an agent for dispersing and / or dissolving the drug in the ointment base, which is selected from poly(ethylene glycol), polyethoxylated castor oil, an alcohol having 12 to 20 carbon atoms, and a mixture of two or more of the foregoing components.

[0752] In some embodiments, the ointment base comprises an ophthalmologically acceptable oil and fat base, such as natural waxes, for example white and yellow beeswax, carnauba wax, wool wax (lanolin), purified lanolin, anhydrous lanolin; petroleum waxes, such as paraffin wax, microcrystalline wax; hydrocarbons, such as liquid paraffin, white and yellow soft paraffin, white petrolatum, yellow petrolatum; or combinations thereof.

[0753] The above-mentioned petroleum and fat bases are described in more detail, for example, in the British Pharmacopoeia 2001 edition or the European Pharmacopoeia, 3rd edition.

[0754] In some embodiments, based on the total weight of the composition, the amount of the ointment base present is from about 50% to about 95%, preferably 70% to 90% (by weight).

[0755] Preferred ointment bases include a combination of one or more natural waxes (such as those described above, preferably wool wax (lanolin)) and one or more hydrocarbons (such as those described above, preferably soft paraffin or petrolatum), more preferably in combination with liquid paraffin.

[0756] Particular examples of the above-mentioned ointment bases comprise, for example, 5 to 17 parts by weight of lanolin, and 50 to 65 parts by weight of white petrolatum, and 20 to 30 parts by weight of liquid paraffin.

[0757] In some embodiments, the agent for dispersing and / or dissolving the ophthalmic drug in the ointment base is selected from poly(ethylene glycol), polyethoxylated castor oil, alcohols having 12 to 20 carbon atoms, and mixtures of two or more of said components. The agent is preferably used in an amount of 1% to 20%, more preferably 1% to 10% (by weight) of the total semi-solid composition.

[0758] Alcohols having 12 to 20 carbon atoms specifically include stearyl alcohol (C 18 H 37 OH), cetyl alcohol (C 16 H 33 OH) and mixtures thereof. Preferred is so-called cetearyl alcohol, consisting essentially of stearyl alcohol and cetyl alcohol and preferably containing not less than 40% (by weight) of stearyl alcohol and at least 90% (by weight) of the total amount of stearyl alcohol and cetyl alcohol, and a solid alcohol mixture, and a composition containing not less than 80% (by weight) of cetyl alcohol and an emulsifier, in particular sodium cetearyl sulfate and / or sodium lauryl sulfate, preferably in an amount of not less than 7% (by weight) of the emulsifier.

[0759] Polyethoxylated castor oil is the reaction product of natural or hydrogenated castor oil and ethylene glycol. In some cases, such products are obtained in a known manner, for example by the reaction of natural or hydrogenated castor oil or its fractions with ethylene oxide, for example in a molar ratio of about 1:30 to about 1:60, and optionally removing the free polyethylene glycol component from the product, for example according to the methods disclosed in German Auslegeschriften 1,182,388 and 1,518,819. Particularly suitable and preferred is the commercially available product under the trade name EL, which has a molecular weight (by vapor osmometry) = about 1630, saponification value = about 65 to 70, acid value = about 2, iodine value = about 28 to 32 and nD 25 = about 1.471. Also suitable for use in this category is, for example, HCO-60, a reaction product of hydrogenated castor oil and ethylene oxide, which has the following characteristics: acid value = about 0.3; saponification value = about 47.4; hydroxyl value = about 42.5. pH (5%) = about 4.6; color value = about 40; melting point = about 36.0 °C; freezing point = about 32.4 °C; H2O content (%, KF) = about 0.03.

[0760] According to the present disclosure, in some embodiments, poly(ethylene glycol) is used as an agent for dispersing and / or dissolving ophthalmic drugs in an ointment matrix. Suitable poly(ethylene glycols) are generally mixtures of polymeric compounds of the general formula H—(OCH2—CH2) n OH, where the exponent n generally ranges from 4 to 230, and the average molecular weight is generally from about 200 to about 10,000. In some embodiments, n is a number from about 6 to about 22, and the average molecular weight is between about 300 and about 1000. In some embodiments, n ranges from about 6 to about 13, and the average molecular weight is from about 300 to about 600. In some embodiments, the value of n is from about 8.5 to about 9, and the relative molecular weight is about 400. Suitable poly(ethylene glycols) are readily commercially available, for example, poly(ethylene glycols) with an average molecular weight of about 200, 300, 400, 600, 1000, 1500, 2000, 3000, 4000, 6000, 8000 and 10,000.

[0761] In some embodiments, poly(ethylene glycol), especially of the type described in the above paragraph, is used in an amount of 1% to 10%, or 1% to 5% (by weight) of the total semi-solid composition.

[0762] In some embodiments, the composition comprises an agent for dispersing and / or dissolving a drug in an ointment matrix, the agent being selected from poly(ethylene glycol), polyethoxylated castor oil and preferably a mixture of said components.

[0763] Gel / ointment viscosity

[0764] In some embodiments, the composition has a Brookfield RVDV viscosity of about 10,000 to about 300,000 cps at about 20 °C and a shear rate of 1 s -1 In some embodiments, the composition has a Brookfield RVDV viscosity of about 15,000 to about 200,000 cps at about 20 °C and a shear rate of 1 s -1 In some embodiments, the composition has a Brookfield RVDV viscosity of about 50,000 to about 150,000 cps at about 20 °C and a shear rate of 1 s -1 In some embodiments, the composition has a Brookfield RVDV viscosity of about 70,000 to about 130,000 cps at about 20 °C and a shear rate of 1 s -1 In some embodiments, the composition has a Brookfield RVDV viscosity of about 90,000 to about 110,000 cps at about 20 °C and a shear rate of 1 s -1 .

[0765] In some embodiments, the ophthalmic gel composition contains a thickening agent sufficient to provide a viscosity between about 500 and 1,000,000 centipoise, between about 750 and 1,000,000 centipoise; between about 1000 and 1,000,000 centipoise; between about 1000 and 400,000 centipoise; between about 2000 and 100,000 centipoise; between about 3000 and 50,000 centipoise; between about 4000 and 25,000 centipoise; between about 5000 and 20,000 centipoise; or between about 6000 and 15,000 centipoise. In some embodiments, the ophthalmic gel composition contains a thickening agent sufficient to provide a viscosity of about 500,000 to 1,000,000 centipoise

[0766] In some embodiments, the compositions described herein are low-viscosity compositions at body temperature. In some embodiments, the low-viscosity composition contains from about 1% to about 10% thickening agent (e.g., gelling component such as polyethylene glycol-oxypropylene copolymer). In some embodiments, the low-viscosity composition contains from about 2% to about 10% thickening agent (e.g., gelling component such as polyethylene glycol-oxypropylene copolymer). In some embodiments, the low-viscosity composition contains from about 5% to about 10% thickening agent (e.g., gelling component such as polyethylene glycol-oxypropylene copolymer). In some embodiments, the low-viscosity composition is substantially free of thickening agent (e.g., gelling component such as polyethylene glycol-oxypropylene copolymer). In some embodiments, the low-viscosity ophthalmic pharmaceutical composition described herein provides an apparent viscosity of from about 100 cps to about 10,000 cps. In some embodiments, the low-viscosity ophthalmic pharmaceutical composition described herein provides an apparent viscosity of from about 500 cps to about 10,000 cps. In some embodiments, the low-viscosity ophthalmic pharmaceutical composition described herein provides an apparent viscosity of from about 1000 cps to about 10,000 cps.

[0767] In some embodiments, the compositions described herein are viscous compositions at body temperature. In some embodiments, the viscous composition contains from about 10% to about 25% thickening agent (e.g., gelling component such as polyethylene glycol-oxypropylene copolymer). In some embodiments, the viscous composition contains from about 14% to about 22% thickening agent (e.g., gelling component such as polyethylene glycol-oxypropylene copolymer). In some embodiments, the viscous composition contains from about 15% to about 21% thickening agent (e.g., gelling component such as polyethylene glycol-oxypropylene copolymer). In some embodiments, the viscous composition described herein provides an apparent viscosity of from about 100,000 cps to about 1,000,000 cps. In some embodiments, the viscous composition described herein provides an apparent viscosity of from about 150,000 cps to about 500,000 cps. In some embodiments, the viscous composition described herein provides an apparent viscosity of from about 250,000 cps to about 500,000 cps. In some such embodiments, the viscous composition is a liquid at room temperature and a gel between room temperature and body temperature (including individuals with severe fever, e.g., up to about 42 °C). In some embodiments, the viscous composition is administered as a monotherapy to treat the ophthalmic diseases or disorders described herein.

[0768] In some embodiments, the viscosity of the gel compositions described herein is measured by any of the described methods. For example, in some embodiments, an LVDV-II + CP cone and plate viscometer and a CPE-40 type conical rotor are used to calculate the viscosity of the gel compositions described herein. In other embodiments, a Brookfield (rotor and measuring cup) viscometer is used to calculate the viscosity of the gel compositions described herein. In some embodiments, the viscosity ranges described herein are measured at room temperature. In other embodiments, the viscosity ranges described herein are measured at body temperature (e.g., at the average body temperature of a healthy person).

[0769] Gel / ointment dose-dose uniformity

[0770] Typical ophthalmic gels are packaged in eye drop bottles and administered as drops. For example, a single administration (i.e., single dose) of an ophthalmic gel includes dropping one, two, three or more drops into a patient's eye. Additionally, typical ophthalmic ointments are packaged in tubes or other squeezable containers with a dispensing nozzle through which a strip of ointment is delivered. For example, a single administration (i.e., single dose) of an ophthalmic ointment includes administering a single strip or multiple strips to a patient's eye. In some embodiments, one dose of the ophthalmic gel described herein is one drop of the gel composition in an eye drop bottle. In some embodiments, one dose of the ophthalmic ointment is one strip of the ointment composition dispensed through the nozzle of a dispensing tube.

[0771] In some cases, the ophthalmic gel compositions described herein provide dose-to-dose uniform concentration. In some cases, there is no significant change in the drug content from one dose to another in the dose-to-dose uniform concentration. In some cases, the dose-to-dose uniform concentration provides a consistent drug content from one dose to another.

[0772] In some cases, the ophthalmic ointment compositions described herein provide dose-to-dose uniform concentration. In some cases, there is no significant change in the drug content from one dose to another in the dose-to-dose uniform concentration. In some cases, the dose-to-dose uniform concentration provides a consistent drug content from one dose to another.

[0773] In some embodiments, the change in the dose-to-dose ophthalmic pharmaceutical concentration of the composition is less than 50%. In some embodiments, the change in the dose-to-dose ophthalmic pharmaceutical concentration of the composition is less than 40%. In some embodiments, the change in the dose-to-dose ophthalmic pharmaceutical concentration of the composition is less than 30%. In some embodiments, the change in the dose-to-dose ophthalmic pharmaceutical concentration of the composition is less than 20%. In some embodiments, the change in the dose-to-dose ophthalmic pharmaceutical concentration of the composition is less than 10%. In some embodiments, the change in the dose-to-dose ophthalmic pharmaceutical concentration of the composition is less than 5%.

[0774] In some embodiments, the dose-to-dose ophthalmic agent concentration variation is based on 10 consecutive doses. In some embodiments, the dose-to-dose ophthalmic agent concentration variation is based on 8 consecutive doses. In some embodiments, the dose-to-dose ophthalmic agent concentration variation is based on 5 consecutive doses. In some embodiments, the dose-to-dose ophthalmic agent concentration variation is based on 3 consecutive doses. In some embodiments, the dose-to-dose ophthalmic agent concentration variation is based on 2 consecutive doses.

[0775] The precipitate-free composition should not require shaking to disperse the drug evenly. The "non-shaking" composition may be advantageous compared to compositions that require shaking simply because the patient's shaking behavior is a major source of variability in the drug dosage administered. It has been reported that patients generally do not or forget to shake the composition before administering the dose, even though it is clearly labeled to indicate shaking on the label. On the other hand, even for those patients who do shake the product, it is generally not possible to determine whether the shaking intensity and / or duration is sufficient to homogenize the product. In some embodiments, the ophthalmic gel compositions and ophthalmic ointment compositions described herein are "non-shaking" compositions that maintain the dose-to-dose uniformity described herein.

[0776] To evaluate dose-to-dose uniformity, the dropper bottle or tube containing the ophthalmic aqueous composition, ophthalmic gel composition, or ophthalmic ointment composition is placed upright until 12 hours before the start of the test. To simulate the recommended doses of these products, a predetermined number of drops or strips are dispensed from each commercially available bottle or tube at predetermined time intervals, or until there is no product left in the bottle or tube. All drops and strips are dispensed into tared glass vials, capped, and stored at room temperature until analysis. The concentration of the double-stranded DNA-binding moiety in the expressed droplets is determined using a reverse-phase HPLC method.

[0777] Treatment method

[0778] In one aspect, the present disclosure provides methods of treating an eye disorder in an individual in need thereof. In some embodiments, the method comprises administering a double-stranded deoxyribonucleic acid (DNA) binding moiety to the eye. In some embodiments, systemic exposure to the double-stranded DNA binding moiety is minimal. In some embodiments, the double-stranded DNA binding moiety binds to a repeat sequence. In some embodiments, the repeat sequence does not contain multiple copies of GAA. In some embodiments, the repeat sequence comprises a repeat of at least four nucleotides. In some embodiments, the repeat sequence comprises multiple copies of CTG, TGC, CAG, GGC, TCCAA, GGG GCC, CGG, GCC, GCG, GCT, GCA, GAC, AAGGG, ATTCT, TGGAA, GGCCTG, CCCCCGCCCCGCG, or CCTG. In some embodiments, the repeat sequence comprises multiple copies of CTG or CAG. In some embodiments, the repeat sequence is not a triplet repeat. In some embodiments, the double-stranded DNA binding moiety is suitable for treating a genetic disease. In some embodiments, the genetic disease is not Friedreich's ataxia. In some embodiments, the double-stranded DNA binding moiety is a polymer that does not contain nucleotides or amino acids. In some embodiments, the double-stranded DNA binding moiety binds to the minor groove of double-stranded DNA. In some embodiments, the double-stranded DNA binding moiety is not an antibiotic.

[0779] In aspects of the treatment methods provided herein, the double-stranded DNA binding moiety binds to a sequence (such as a repeat sequence) adjacent to or within a gene. In some embodiments, the double-stranded DNA binding moiety regulates the expression of the gene. In some embodiments, the double-stranded DNA binding moiety increases the expression of the gene. In some embodiments, the double-stranded DNA binding moiety decreases the expression of the gene. In some embodiments, the gene is transcription factor 4 (TCF4).

[0780] In aspects of the treatment methods provided herein, the double-stranded DNA binding moiety binds non-covalently to double-stranded DNA. In some embodiments, the double-stranded DNA binding moiety binds to double-stranded DNA using electrostatic forces (such as hydrogen bonds or van der Waals forces). In some embodiments, the double-stranded DNA binding moiety does not contain polynucleotides. In some embodiments, the double-stranded DNA binding moiety does not contain polypeptides.

[0781] In aspects of the therapeutic methods provided herein, the molecular weight of the double-stranded DNA-binding moiety is less than about 15 kDa, less than about 12.5 kDa, less than about 10 kDa, less than about 9.5 kDa, less than about 9 kDa, less than about 8.5 kDa, less than about 8 kDa, less than about 7.5 kDa, less than about 7 kDa, less than about 6.5 kDa, less than about 6 kDa, less than about 5.5 kDa, less than about 5 kDa, less than about 4.5 kDa, less than about 4 kDa, less than about 3.5 kDa, less than about 3 kDa, less than about 2.5 kDa, less than about 2 kDa or less than about 1.5 kDa. In some embodiments, the molecular weight of the double-stranded DNA-binding moiety is less than about 10 kDa, about 7.5 kDa, about 5 kDa or about 2.5 kDa.

[0782] In some embodiments, the double-stranded DNA-binding moiety comprises a polyamide. In some embodiments, the double-stranded DNA-binding moiety further comprises a transcriptional regulator-binding moiety. In some embodiments, the double-stranded DNA-binding moiety is linked to the transcriptional regulator-binding moiety via a linker.

[0783] In aspects of the therapeutic methods provided herein, the individual is a mammal. In some embodiments, the individual is a mouse, rat, cat, dog, rabbit, horse, cow, pig or human. In some embodiments, the individual is a mammal.

[0784] In aspects of the therapeutic methods provided herein, the double-stranded DNA-binding moiety is formulated in a composition comprising an excipient suitable for ocular administration. In some embodiments, the excipient includes an isotonicity regulator (such as sodium chloride), buffer, stabilizer, antioxidant, thickening agent, solubility enhancer, permeation enhancer and / or preservative. In some embodiments, the pH of the composition is from about 5 to about 8. In some embodiments, the viscosity of the composition at about 20 °C is from about 1 to about 50,000 cps.

[0785] In various aspects, administration of the composition to the eye results in minimal systemic exposure. In some embodiments, topical ocular administration (e.g., ophthalmic administration) of the composition results in penetration of the double-stranded DNA-binding moiety into the inner cells and tissues of the eye. In some embodiments, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 95% or at least about 99% of the double-stranded DNA-binding moiety administered to the eye can penetrate the cells and tissues inside the eye.

[0786] In another aspect, the present disclosure provides methods of treating eye disorders in an individual in need thereof, the method comprising administering a double-stranded DNA-binding moiety, wherein the double-stranded DNA-binding moiety is a polymer that does not contain nucleotides or amino acids, and wherein the composition is suitable for administration to the human eye. In some embodiments, the double-stranded DNA-binding moiety does not bind to a repeat sequence comprising multiple copies of GAA. In some embodiments, the double-stranded DNA-binding moiety binds to a repeat sequence comprising multiple copies of CTG, TGC, CAG, GGC, TCCAA, GGG GCC, CGG, GCC, GCG, GCT, GCA, GAC, AAGGG, ATTCT, TGGAA, GGCCTG, CCCCCGCCCCGCG or CCTG. In some embodiments, the repeat sequence comprises multiple copies of CTG or CAG. In some embodiments, the repeat sequence comprises at least four nucleotides. In some embodiments, the repeat sequence is not a triplet repeat.

[0787] In the context of a method of treatment with a double-stranded DNA-binding moiety that does not contain nucleotides or amino acids, the disorder is a genetic disease. In some embodiments, the disorder is not Friedreich's ataxia. In some embodiments, the double-stranded DNA-binding moiety is not an antibiotic. In some embodiments, the double-stranded DNA-binding moiety binds to the minor groove of double-stranded DNA.

[0788] In the context of the methods of treatment provided herein, the double-stranded DNA-binding moiety that does not contain nucleotides or amino acids binds to sequences (such as repeat sequences) adjacent to or within a gene. In some embodiments, the double-stranded DNA-binding moiety regulates gene expression. In some embodiments, the double-stranded DNA-binding moiety increases gene expression. In some embodiments, the double-stranded DNA-binding moiety decreases gene expression. In some embodiments, the gene is transcription factor 4 (TCF4).

[0789] In various aspects of the methods of treatment provided herein, the double-stranded DNA-binding moiety that does not contain nucleotides or amino acids binds non-covalently to double-stranded DNA. In some embodiments, the double-stranded DNA-binding moiety binds to double-stranded DNA using electrostatic forces (such as hydrogen bonds or van der Waals forces). In some embodiments, the double-stranded DNA-binding moiety does not contain polynucleotides. In some embodiments, the double-stranded DNA-binding moiety does not contain polypeptides.

[0790] In aspects of the therapeutic methods provided herein, the molecular weight of the double-stranded DNA-binding moiety that does not contain nucleotides or amino acids is less than about 15 kDa, less than about 12.5 kDa, less than about 10 kDa, less than about 9.5 kDa, less than about 9 kDa, less than about 8.5 kDa, less than about 8 kDa, less than about 7.5 kDa, less than about 7 kDa, less than about 6.5 kDa, less than about 6 kDa, less than about 5.5 kDa, less than about 5 kDa, less than about 4.5 kDa, less than about 4 kDa, less than about 3.5 kDa, less than about 3 kDa, less than about 2.5 kDa, less than about 2 kDa, or less than about 1.5 kDa. In some embodiments, the molecular weight of the double-stranded DNA-binding moiety is less than about 10 kDa, about 7.5 kDa, about 5 kDa, or about 2.5 kDa.

[0791] In some embodiments, the double-stranded DNA-binding moiety that does not contain nucleotides or amino acids comprises a polyamide. In some embodiments, the double-stranded DNA-binding moiety further comprises a transcriptional regulator-binding moiety. In some embodiments, the double-stranded DNA-binding moiety is linked to the transcriptional regulator-binding moiety via a linker.

[0792] In aspects of the therapeutic method, the double-stranded DNA-binding moiety does not contain the nucleotides or amino acids provided herein, and the individual is a mammal. In some embodiments, the individual is a mouse, rat, cat, dog, rabbit, horse, cow, pig, or human. In some embodiments, the individual is a mammal.

[0793] In aspects of the therapeutic method provided herein, the double-stranded DNA-binding moiety that does not contain nucleotides or amino acids is formulated in a composition comprising an excipient suitable for ocular administration. In some embodiments, the excipient includes an isotonicity regulator (such as sodium chloride), a buffer, a stabilizer, an antioxidant, a thickening agent, a solubility enhancer, a permeation enhancer, and / or a preservative. In some embodiments, the pH of the composition is from about 5 to about 8. In some embodiments, the viscosity of the composition at about 20 °C is from about 1 to about 50,000 cps.

[0794] In various aspects, administration of a composition comprising a double-stranded DNA-binding moiety that does not contain nucleotides or amino acids to the eye results in minimal systemic exposure. In some embodiments, topical ocular administration (e.g., ophthalmic administration) of the composition results in penetration of the double-stranded DNA-binding moiety into the internal cells and tissues of the eye. In some embodiments, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or at least about 99% of the double-stranded DNA-binding moiety administered to the eye can penetrate the cells and tissues inside the eye.

[0795] In another aspect, the present disclosure provides methods for treating an eye disorder in an individual in need thereof, the method comprising administering a double-stranded DNA-binding moiety, wherein the eye disorder is a genetic disease. In some embodiments, the genetic disease is not Friedreich's ataxia. In some embodiments, the double-stranded DNA-binding moiety is not an antibiotic. In some embodiments, the double-stranded DNA-binding moiety does not comprise nucleotides or amino acids.

[0796] In some embodiments of the methods for treating an eye disorder, wherein the eye disorder is a genetic disease, the double-stranded DNA-binding moiety does not bind to a repeat sequence comprising multiple copies of GAA. In some embodiments, the double-stranded DNA-binding moiety binds to a repeat sequence comprising multiple copies of CTG, TGC, CAG, GGC, TCCAA, GGG GCC, CGG, GCC, GCG, GCT, GCA, GAC, AAGGG, ATTCT, TGGAA, GGCCTG, CCCCCGCCCCGCG, or CCTG. In some embodiments, the repeat sequence comprises multiple copies of CTG or CAG. In some embodiments, the repeat sequence comprises at least four nucleotides. In some embodiments, the repeat sequence is not a triplet repeat. In some embodiments, the double-stranded DNA-binding moiety binds to the minor groove of double-stranded DNA.

[0797] In an aspect of the methods for treating an eye disorder that is a genetic disease provided herein, the double-stranded DNA-binding moiety binds to a sequence (such as a repeat sequence) adjacent to or within a gene. In some embodiments, the double-stranded DNA-binding moiety regulates the expression of a gene. In some embodiments, the double-stranded DNA-binding moiety increases the expression of a gene. In some embodiments, the double-stranded DNA-binding moiety decreases the expression of a gene. In some embodiments, the gene is transcription factor 4 (TCF4).

[0798] In an aspect of the methods for treating an eye disorder that is a genetic disease provided herein, the double-stranded DNA-binding moiety binds non-covalently to double-stranded DNA. In some embodiments, the double-stranded DNA-binding moiety binds to double-stranded DNA using electrostatic forces (such as hydrogen bonds or van der Waals forces). In some embodiments, the double-stranded DNA-binding moiety does not comprise polynucleotides. In some embodiments, the double-stranded DNA-binding moiety does not comprise polypeptides.

[0799] Provided herein are aspects of methods of treating ophthalmic disorders of genetic diseases, wherein the double-stranded DNA binding moiety has a molecular weight of less than about 15 kDa, less than about 12.5 kDa, less than about 10 kDa, less than about 9.5 kDa, less than about 9 kDa, less than about 8.5 kDa, less than about 8 kDa, less than about 7.5 kDa, less than about 7 kDa, less than about 6.5 kDa, less than about 6 kDa, less than about 5.5 kDa, less than about 5 kDa, less than about 4.5 kDa, less than about 4 kDa, less than about 3.5 kDa, less than about 3 kDa, less than about 2.5 kDa, less than about 2 kDa or less than about 1.5 kDa. In some embodiments, the double-stranded DNA binding moiety has a molecular weight of less than about 10 kDa, about 7.5 kDa, about 5 kDa or about 2.5 kDa.

[0800] In some embodiments of methods of treating ophthalmic disorders of genetic diseases, the double-stranded DNA binding moiety comprises a polyamide. In some embodiments, the double-stranded DNA binding moiety further comprises a transcriptional regulator binding moiety. In some embodiments, the double-stranded DNA binding moiety is linked to the transcriptional regulator binding moiety via a linker.

[0801] Provided herein are aspects of methods of treating ophthalmic disorders of genetic diseases, wherein the individual is a mammal. In some embodiments, the individual is a mouse, rat, cat, dog, rabbit, horse, cow, pig or human. In some embodiments, the individual is a mammal.

[0802] Provided herein are aspects of methods of treating ophthalmic disorders of genetic diseases, wherein the double-stranded DNA binding moiety is formulated in a composition comprising an excipient suitable for ocular administration. In some embodiments, the excipient includes an isotonicity regulator (such as sodium chloride), buffer, stabilizer, antioxidant, thickening agent, solubility, permeation enhancer and / or preservative. In some embodiments, the pH of the composition is from about 5 to about 8. In some embodiments, the viscosity of the composition at about 20 °C is from about 1 to about 50,000 cps.

[0803] In various aspects of methods of treating ophthalmic disorders of genetic diseases, administration of the composition to the eye results in minimal systemic exposure. In some embodiments, topical ocular administration (e.g., ophthalmic administration) of the composition results in penetration of the double-stranded DNA binding moiety into the inner cells and tissues of the eye. In some embodiments, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 95% or at least about 99% of the topically administered double-stranded DNA binding moiety can penetrate the inner cells and tissues of the eye.

[0804] In various aspects of the treatment methods provided herein, tissue biodistribution can be evaluated in single-dose and multi-dose ophthalmic administration studies that can be conducted in experimental models such as rats, rabbits, and dogs. In some cases, after ophthalmic administration, the levels of the double-stranded DNA-binding agent in ocular tissues consistently show an exposure gradient, with the highest concentration of the double-stranded DNA-binding agent measured in the outer eye structures closest to the administration site, gradually decreasing to the innermost corneal endothelium, which is the target tissue for the double-stranded DNA-binding agent.

[0805] In some cases, after repeated dosing for about 14 days (e.g., 1 to 28 days, 7 to 28 days, 14 to 28 days, 21 to 28 days, 1 to 21 days, 7 to 21 days, 14 to 21 days, 1 to 14 days, 7 to 14 days, or 1 to 7 days (once or twice a day)) up to 24 hours (e.g., 2 to 24 hours, 6 to 24 hours, 12 to 24 hours, 18 to 24 hours, 2 to 18 hours, 6 to 18 hours, 12 to 18 hours, 2 to 12 hours, 6 to 12 hours, or 2 to 6 hours), the double-stranded DNA-binding agent can be detected in the corneal endothelium at a concentration equal to or exceeding 3 to 20 nM (e.g., at least 3 nM, at least 6 nM, at least 9 nM, at least 12 nM, at least 15 nM, at least 18 nM, at least 20 nM, or more), and this concentration is predicted to alleviate symptoms such as the number of nuclear foci and instances of splicing abnormalities in corneal endothelial cells (CECs) of some diseased individuals. In one embodiment, after the final ophthalmic administration, e.g., within 10 minutes, 20 minutes, 30 minutes, 40 minutes, 50 minutes, 1 hour, 1.2 hours, 1.4 hours, 1.6 hours, 1.8 hours, etc., the endothelial concentration is up to 10-fold, 20-fold, 30-fold, 40-fold, 50-fold, 60-fold, 80-fold, 100-fold, or 120-fold more than the lowest or highest predicted effective concentration. In some cases, effective concentrations can be achieved when the double-stranded DNA-binding agent is administered once a day over several days, weeks, or months using a clinically relevant dosing paradigm. In some cases, effective concentrations can be achieved when the double-stranded DNA-binding agent is administered twice a day over several days, weeks, or months using a clinically relevant dosing paradigm. In some cases, the results of distribution studies using aqueous formulations (such as acidified aqueous formulations) indicate that ophthalmic administration results in a concentration of the double-stranded DNA-binding agent in the target endothelium that exceeds the predicted biologically relevant concentration at about 24 hours after dosing.

[0806] In various aspects of the treatment methods provided herein, aqueous formulations (such as acidified aqueous formulations) optionally contain mannitol, such as selected from 1% to 10% mannitol, for example 2% to 10%, 2% to 8%, 2% to 6%, 2% to 4%, 4% to 10%, 4% to 8%, 4% to 6%, 6% to 10%, 6% to 8% or 8% to 10% as an ophthalmic formulation of a double-stranded DNA binder. In some cases, in GLP studies in rabbits and dogs for about 4 weeks (such as 1 to 4 weeks, 2 to 4 weeks or 1 to 2 weeks), an ocular dosing regimen (e.g., four drops of 50 μL each, such as 1, 2, 3, 4, 5 or 6 drops, 10 μL, 20 μL, 30 μL, 40 μL, 50 μL, 60 μL, 70 μL, 80 μL, 90 μL or 100 μL, once, twice or three times a day) and concentrations (0.6% and 1%, such as 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9% or 2%) were used. The double-stranded DNA binder formulated in this way was well tolerated locally after about 7 days of daily administration (such as 1 to 14 days, 1 to 10 days, 1 to 8 days, 1 to 6 days, 1 to 4 days, 1 to 2 days, 2 to 14 days, 2 to 10 days, 2 to 8 days, 2 to 6 days, 2 to 4 days, 4 to 14 days, 4 to 10 days, 4 to 8 days, 4 to 6 days, 6 to 14 days, 6 to 10 days, 6 to 8 days, 8 to 14 days, 8 to 10 days or 10 to 14 days). In some cases, the ocular target tissue levels were equivalent to those observed in earlier studies using acidified water alone. In some cases, the highest exposure of the double-stranded DNA binder was observed in the corneal endothelium within the first hours after topical instillation, such as 20 minutes, 40 minutes, 1 hour after treatment. In the next 2 to 24 hours (such as 2 hours, 4 hours, 6 hours, 8 hours, 10 hours, 12 hours, 14 hours, 16 hours, 18 hours, 20 hours, 22 hours or 24 hours), the level of the double-stranded DNA binder in the corneal endothelium then decreased to about 3 to 40 nM (such as 3 nM, 6 nM, 8 nM, 10 nM, 12 nM, 14 nM, 16 nM, 18 nM, 20 nM, 24 nM, 28 nM, 32 nM, 36 nM or 40 nM), indicating the diffusion and / or transport of the double-stranded DNA binder in the cornea. In some cases, the double-stranded DNA binder was detected in the aqueous humor up to 12 to 48 hours after the last dose, for example 12 hours, 16 hours, 20 hours, 24 hours, 28 hours, 32 hours, 36 hours, 40 hours, 44 hours or 48 hours after the last dose.In some cases, the total volume of aqueous humor in a study animal (such as a rabbit) is replenished approximately every 1 to 4 hours (such as 1 hour, 2 hours, 3 hours, or 4 hours), so at later time points, the level of double-stranded DNA-binding agent measurable in the aqueous humor indicates release of the double-stranded DNA-binding agent from the cornea into the posterior chamber of the eye.

[0807]

[0808]

[0809]

[0810]

[0811] Examples

[0812] The following examples are given to illustrate various embodiments of the invention and are not meant to limit the invention in any way. The present examples and the methods described herein currently represent preferred embodiments, are exemplary, and are not intended to limit the scope of the invention. Those skilled in the art will envision variations and other uses that fall within the spirit of the invention as defined by the scope of the claims.

[0813] Example 1: Determination of the solubility of a compound in water

[0814] To determine the solubility of a candidate compound, in-situ salt formation was used with an aqueous hydrochloric acid solution to determine the solubility of the compound at a concentration of 10 mg / ml. First, 0.5 ml of sterile water was added to 10 mg of the dry compound, vortexed, and briefly centrifuged. Next, 0.5 equivalents of 0.025 M HCl was added to the mixture, and then water was added to bring the total volume to 1 ml. The mixture was continuously stirred for 0.5 to 24 hours. If a clear solution was obtained, the pH value was determined using a microelectrode. If the solution was not clear, additional water and / or HCl was added, and the mixture was continuously stirred and vortexed until it was determined that the compound was either completely dissolved or insoluble. The minimum concentration tested was 1 mg / ml. The pH value was evaluated at the end of the experiment. The experimental results are shown in Table 2

[0815]

[0816]

[0817] Example 2: Determination of compound activity using an in vitro focus assay

[0818] Sixteen hours before compound treatment, immortalized Fuchs corneal endothelial dystrophy cells (F35T) were seeded at 5000 cells per well in a 96-well black-sided clear-bottom plate. The compound was dissolved in DMSO to 1 mM and diluted in F35T cell medium to a final concentration of 3000 to 1 nM. The diluted compound was added to the F35T cells and incubated for 48 hours.

[0819] After treatment, the cells were fixed in 4% paraformaldehyde for 20 minutes, washed twice with PBS, and permeabilized with 70% ethanol at -20 °C for 16 hours. After permeabilization, the cells were washed twice with PBS and then hydrated with 30% formamide plus 2x SSC buffer for 10 minutes. Nuclei foci were labeled by fluorescence in situ hybridization with (CAG)10-Cy3 oligonucleotide. Images were collected and automated image analysis was performed on a BioTEK Cytation 5. The experimental data are provided in Table 3.

[0820]

[0821] Example 3: Evaluation of ocular penetration of a compound formulated in water

[0822] As previously described, freshly enucleated bovine eyes (Nebraska Scientific) were treated with the compounds (Luschmann et al., “Developing an in situ nanosuspension: a novel approach towards the efficient administration of poorly soluble drugs at the anterior eye”, European Journal of Pharmaceutical Sciences (2013)). Individual eyes were rinsed with 10 mL of sterile 0.9% saline and then placed at the bottom of a 600 mL glass beaker (Pyrex). A borosilicate glass powder funnel (Eisco Labs) with a diameter of 26 mm was gently placed over the eye such that the cornea was centered in the cylindrical opening, creating a sealed dosing chamber in which the solution was administered. The eyes were equilibrated in sterile 0.9% saline at room temperature for 20 minutes and then the saline was poured off. 500 μL of a 0.1% to 0.3% GeneTAC compound was applied to the cornea, followed by 167 μL of sterile water to mimic a 30 μL eye drop instilled into 10 μL of tear fluid. The compound was incubated at room temperature for 30 minutes and then the funnel was removed. Next, the eyes were washed twice with 10 mL of a 5.2% mannitol aqueous solution to remove residual compound. Aqueous humor was recovered using a 28-gauge needle and frozen in liquid nitrogen. The cornea was then surgically excised and the Descemet's membrane and corneal endothelium (DM / CE) were removed with fine forceps under a dissecting microscope and frozen in liquid nitrogen. Compound abundance was quantified in aqueous humor, anterior cornea, and DM / CE using liquid chromatography mass spectrometry. The data are shown in Table 4 below. The data are also shown in Figure 1 which shows that compound 201 reaches concentrations in the corneal epithelium and stroma and in the Descemet's membrane and endothelium that exceed the IC50 for preventing focus formation and exceed the IC50 for preventing aberrant splicing events.

[0823]

[0824] While the preferred embodiments of the present invention have been shown and described herein, it will be apparent to those skilled in the art that such embodiments are provided by way of example only. Many variations, changes, and substitutions will now occur to those skilled in the art without departing from the invention. It should be understood that various alternatives to the embodiments described herein may be employed. The following claims are intended to define the scope of the invention and thereby cover the methods and structures within the scope of these claims and their equivalents.

Claims

1. A composition comprising a double-stranded DNA-binding moiety that binds to double-stranded deoxyribonucleic acid (DNA) in a sequence-specific manner, wherein the double-stranded DNA-binding moiety does not bind to a repetitive sequence comprising multiple copies of GAA, and wherein the composition is suitable for administration to the human eye.

2. A composition comprising a double-stranded DNA-binding moiety that is suitable for treating a genetic disease, wherein the composition is suitable for administration to the human eye.

3. A composition comprising a double-stranded DNA-binding moiety that binds to a repetitive sequence comprising at least four nucleotides, wherein the composition is suitable for administration to the human eye.

4. A composition comprising a double-stranded DNA-binding moiety that binds to a repetitive sequence comprising multiple copies of: CTG, TGC, CAG, GGC, TCCAA, GGGGCC, CGG, GCC, GCG, ATTCT, TGGAA, GGCCTG, CCCCGCCCCGCG or CCTG, wherein the composition is suitable for administration to the human eye.

5. A composition comprising a double-stranded DNA-binding moiety, wherein the double-stranded DNA-binding moiety is a polymer that does not contain nucleotides or amino acids, and wherein the composition is suitable for administration to the human eye.

6. The composition according to any one of claims 1 to 5, wherein the double-stranded DNA-binding moiety binds to a repetitive sequence comprising multiple copies of CTG or CAG.

7. The composition according to any one of claims 1 to 6, wherein the double-stranded DNA-binding moiety binds to the minor groove of double-stranded DNA.

8. The composition according to any one of claims 1 to 7, wherein the double-stranded DNA-binding moiety binds to a repetitive sequence adjacent to or within a gene.

9. The composition according to claim 8, wherein the double-stranded DNA-binding moiety regulates the expression of the gene.

10. The composition according to claim 8 or claim 9, wherein the gene is transcription factor 4 (TCF4).

11. The composition according to any one of claims 1 to 10, wherein the double-stranded DNA-binding moiety binds non-covalently.

12. The composition according to any one of claims 1 to 4 or 6 to 11, wherein the double-stranded DNA-binding moiety does not contain a polynucleotide.

13. The composition according to any one of claims 1 to 4 or 6 to 12, wherein the double-stranded DNA-binding moiety does not contain a polypeptide.

14. The composition according to any one of claims 1 to 13, wherein the molecular weight of the double-stranded DNA-binding moiety is less than about 10 kDa, about 7.5 kDa, about 5 kDa or about 2.5 kDa.

15. The composition according to any one of claims 1 to 14, wherein the double-stranded DNA-binding moiety comprises a polyamide.

16. The composition according to any one of claims 1 to 15, wherein the double-stranded DNA-binding moiety further comprises a transcription regulator-binding moiety.

17. The composition according to claim 16, wherein the double-stranded DNA binding moiety is linked to the transcriptional regulator binding moiety via a linker.

18. The composition according to any one of claims 1 to 17, wherein administration of the composition to the eye results in minimal systemic exposure.

19. The composition according to any one of claims 1 to 18, which further comprises an excipient.

20. The composition according to any one of claims 1 to 19, wherein the pH of the ophthalmic formulation composition is from about 5 to about 8.

21. The composition according to any one of claims 1 to 20, wherein the composition has a viscosity of from about 1 to about 50,000 cps at about 20 °C.

22. A method of treating an eye disorder in a human subject in need thereof, the method comprising administering a double-stranded DNA binding moiety that binds double-stranded deoxyribonucleic acid (DNA) in a sequence-specific manner, wherein the double-stranded DNA binding moiety does not bind to a repetitive sequence comprising multiple copies of GAA.

23. A method of treating an eye disorder in a human subject in need thereof, the method comprising administering a double-stranded DNA binding moiety, wherein the eye disorder is a genetic disease.

24. A method of treating an eye disorder in a human subject in need thereof, the method comprising administering a double-stranded DNA binding moiety to the eye of the subject, wherein administration results in minimal systemic exposure of the double-stranded DNA binding moiety, and wherein the double-stranded DNA binding moiety binds to a repetitive sequence comprising multiple copies of: CTG, TGC, CAG, GGC, TCCAA, GGG GCC, CGG, GCC, GCG, GCT, GCA, GAC, AAGGG, ATTCT, TGGAA, GG CCTG, CCCCCGCCCCGCG or CCTG.

25. A method of treating an eye disorder in a human subject in need thereof, the method comprising administering a double-stranded DNA binding moiety that binds to a repetitive sequence comprising at least four nucleotides.

26. A method of treating an eye disorder in a human subject in need thereof, the method comprising administering a double-stranded DNA binding moiety, wherein the double-stranded DNA binding moiety is a polymer that does not contain nucleotides or amino acids.

27. The method according to any one of claims 22 to 26, wherein the double-stranded DNA binding moiety binds to a repetitive sequence comprising multiple copies of CTG or CAG.

28. The method according to any one of claims 22 to 27, wherein the double-stranded DNA binding moiety binds to the minor groove of double-stranded DNA.

29. The method according to any one of claims 22 to 28, wherein the double-stranded DNA binding moiety binds to a repetitive sequence adjacent to or within a gene.

30. The method according to claim 29, wherein the double-stranded DNA binding moiety regulates the expression of the gene.

31. The method according to claim 29 or claim 30, wherein the gene is transcription factor 4 (TCF4).

32. The method according to any one of claims 22 to 31, wherein the double-stranded DNA binding moiety binds non-covalently.

33. The method according to any one of claims 22 to 25 or 27 to 32, wherein the double-stranded DNA binding moiety does not comprise a polynucleotide.

34. The method according to any one of claims 22 to 25 or 27 to 33, wherein the double-stranded DNA binding moiety does not comprise a polypeptide.

35. The method according to any one of claims 22 to 34, wherein the molecular weight of the double-stranded DNA binding moiety is less than about 10 kDa, about 7.5 kDa, about 5 kDa or about 2.5 kDa.

36. The method according to any one of claims 22 to 35, wherein the double-stranded DNA binding moiety comprises a polyamide.

37. The method according to any one of claims 22 to 36, wherein the double-stranded DNA binding moiety further comprises a transcriptional regulator binding moiety.

38. The method according to claim 37, wherein the double-stranded DNA binding moiety is linked to the transcriptional regulator binding moiety via a linker.

39. The method according to claim 22 or 25 to 38, wherein the double-stranded DNA binding moiety is formulated in an excipient suitable for ocular administration.

40. The method according to any one of claims 22 to 39, wherein the double-stranded DNA binding moiety is in a medium having a pH of about 5 to about 8.

41. The method according to any one of claims 22 to 40, wherein the double-stranded DNA binding moiety is in a medium having a viscosity of about 1 to about 50,000 cps at about 20 °C.

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