Methods and compositions for the treatment of ocular diseases

Abstract

Compositions and methods of use for ocular administration are provided herein.

Description

[Technical Field]

[0001] cross reference This application claims the benefit of U.S. Application No. 63 / 380,332, filed October 20, 2022, which is incorporated herein by reference in its entirety. [Background technology]

[0002] Drug delivery to the human eye is an important area of ​​pharmaceutical development. The anatomy and physiology of the eye present challenges in getting drugs to their site of action and maintaining therapeutic efficacy over time. Summary of the Invention

[0003] In one aspect, the present invention provides a composition comprising a double-stranded deoxyribonucleic acid (DNA) binding moiety that binds to double-stranded DNA in a sequence-specific manner.In some embodiments, the double-stranded DNA binding moiety does not bind to the repeat sequence that contains multiple copies of GAA.In some embodiments, the composition is suitable for administration to human eyes.

[0004] In another aspect, provided herein is a composition comprising a double-stranded DNA binding moiety suitable for treating a genetic disease, wherein the composition is suitable for administration to the human eye.

[0005] In a further aspect, provided herein is a composition comprising a double-stranded DNA binding moiety that binds to a repeat sequence comprising at least four nucleotides, wherein the composition is suitable for administration to the human eye.

[0006] A composition comprising a double-stranded DNA binding moiety that binds to a repeat sequence containing 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.

[0007] 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 the composition is suitable for administration to the human eye.

[0008] In various aspects of the compositions provided herein, in some embodiments, the double-stranded DNA-binding moiety binds to a repeat sequence containing 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 regulates expression of a gene. 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 double-stranded DNA-binding moiety has a molecular weight of less than about 10 kDa, less than about 7.5 kDa, less than about 5 kDa, or less than 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 transcription modulator-binding moiety. In some embodiments, the double-stranded DNA-binding moiety is connected to the transcription modulator-binding moiety by 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.

[0009] In another aspect, provided herein is a method for treating an eye disorder in a human individual in need thereof, the method comprising administering a double-stranded deoxyribonucleic acid (DNA) binding moiety that binds to double-stranded DNA in a sequence-specific manner. In some embodiments, the double-stranded DNA binding moiety does not bind to a repetitive sequence that contains multiple copies of GAA.

[0010] In another aspect, provided herein is a method of treating an ocular disorder in a human individual in need thereof, said method comprising administering a double-stranded DNA binding moiety, wherein the ocular disorder is a genetic disease.

[0011] In a further aspect, provided herein is a method of treating an ocular disorder in a human individual in need thereof, said method comprising administering to the eye of the individual a double-stranded DNA-binding moiety, wherein the administration results in minimal systemic exposure to the double-stranded DNA-binding moiety, and wherein the double-stranded DNA-binding moiety binds to a repeat sequence comprising multiple copies of CTG, TGC, CAG, GGC, TCCAA, GGGGCC, CGG, GCC, GCG, GCT, GCA, GAC, AAGGG, ATTCT, TGGAA, GGCCTG, CCCCGCCCCGCG, or CCTG.

[0012] In another aspect, provided herein is a method of treating an ocular disorder in a human individual in need thereof, the method comprising administering a double-stranded DNA-binding moiety that binds to a repeat sequence comprising at least four nucleotides.

[0013] In a further aspect, provided herein is a method of treating an ocular disorder in a human 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.

[0014] In various aspects of the methods herein, in some embodiments, the double-stranded DNA-binding moiety binds to a repeat sequence containing 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 regulates expression of a gene. 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 double-stranded DNA-binding moiety has a molecular weight of less than about 10 kDa, less than about 7.5 kDa, less than about 5 kDa, or less than 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 transcription modulator-binding moiety. In some embodiments, the double-stranded DNA-binding moiety is connected to the transcription modulator-binding moiety by a linker. In some embodiments, the double-stranded DNA-binding moiety is formulated in an excipient suitable for ocular administration. 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.

[0015] Incorporation by Reference All publications, patents, and patent applications mentioned in this specification are herein incorporated 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 explanation of the drawings]

[0016] A better understanding of the features and advantages of the present invention will be obtained by reference to the following detailed description that sets forth illustrative embodiments, in which the principles of the invention are utilized, and the accompanying drawings of which: [Figure 1] 1 shows the ex vivo bovine ocular distribution of Compound 201. DETAILED DESCRIPTION OF THE INVENTION

[0017] In the treatment of eye-affecting diseases, the treatment of eye-affected cells and tissues is required, but systemic administration and exposure are not necessary or desirable.Therefore, in many cases, it is beneficial to directly administer eye-affected treatment to eye, and minimize the exposure to the rest of the body.This feature of eye-affected treatment can sometimes avoid off-target effects, reduce the immune response to pharmaceutical compositions, and ensure that the maximum dose reaches eye-targeted cells and tissues, thereby increasing therapeutic efficacy.

[0018] 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. Direct administration of such compositions to the eye with minimal systemic exposure has not previously been demonstrated, and therefore, the prior art does not demonstrate ocular administration of these compositions.

[0019] Provided herein are compositions for administration to the eye, for example, ophthalmic compositions containing a drug formulated for administration to the eye, and methods for treating eye diseases using such compositions. In some embodiments, compositions are provided that contain a drug that acts by regulating gene expression by binding to double-stranded deoxyribonucleic acid (DNA). In some embodiments, such compositions contain a drug that regulates genes adjacent to a specific DNA sequence by binding to that DNA sequence and recruiting transcriptional regulators, such as transcriptional activators or repressors, thereby regulating the expression of one or more genes. In further embodiments, administering such compositions to the eye allows the therapeutic agent to remain in the eye, minimizing exposure to the rest of the body.

[0020] Double-stranded deoxyribonucleic acid binding composition Provided herein are 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 include multiple copies of GAA. In some embodiments, the repetitive sequence includes repeats of at least four nucleotides. In some embodiments, the repetitive sequence includes 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 includes 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 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 include 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 a human eye. In some embodiments, the double-stranded DNA binding moiety binds to the minor groove of double-stranded DNA.

[0021] In aspects of the compositions 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 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).

[0022] In the embodiments of the compositions provided herein, the double-stranded DNA binding moiety binds to double-stranded DNA non-covalently.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.

[0023] In embodiments of the compositions provided herein, 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, less than about 7.5 kDa, less than about 5 kDa, or less than about 2.5 kDa.

[0024] 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 connected to the transcriptional modulator-binding moiety by a linker.

[0025] In various aspects, the composition is formulated to be suitable for topical administration. In some embodiments, the composition further comprises an excipient. In some embodiments, the excipient comprises an isotonicity adjusting agent, e.g., sodium chloride, a buffer, a stabilizer, an antioxidant, a viscosity enhancing agent, a solubilizing agent, a penetration enhancer, and / or a preservative. In some embodiments, the 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 centipoise (cps) at about 20°C.

[0026] In various embodiments, administration of the composition to the eye results in minimal systemic exposure. In some embodiments, local administration of the composition to the eye (e.g., ocular administration) results in the 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 penetrates the internal cells and tissues of the eye.

[0027] 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 the composition is suitable for administration to the human eye.

[0028] In some embodiments of the nucleotide- or amino acid-free composition, 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, 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 repetitive sequence comprises repeats of at least four nucleotides. In some embodiments, the repetitive sequence does not comprise GAA. In some embodiments, the double-stranded DNA-binding moiety binds to the minor groove of double-stranded DNA.

[0029] In the embodiments of the compositions provided herein that do not contain nucleotides or amino acids, 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.

[0030] In embodiments of the compositions provided herein that do not contain nucleotides or amino acids, 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 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).

[0031] In the embodiments of the compositions provided herein that do not contain nucleotides or amino acids, the double-stranded DNA binding moiety binds to double-stranded DNA non-covalently. 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.

[0032] In embodiments of the compositions provided herein that do not contain nucleotides or amino acids, 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, less than about 7.5 kDa, less than about 5 kDa, or less than about 2.5 kDa.

[0033] 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 modulator-binding moiety. In some embodiments, the double-stranded DNA-binding moiety is connected to the transcriptional modulator-binding moiety by a linker.

[0034] In various embodiments, the composition is suitable for topical administration without containing nucleotides or amino acids. In some embodiments, the composition further comprises an excipient. In some embodiments, the excipient comprises an isotonicity adjuster, e.g., sodium chloride, a buffer, a stabilizer, and an antioxidant, a viscosity enhancer, a solubilizer, a penetration enhancer, and / or a preservative. In some embodiments, the 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.

[0035] In various embodiments, administration of a composition that does not contain nucleotides or amino acids to the eye results in minimal systemic exposure.In some embodiments, local administration of a composition to the eye (e.g., ocular administration) results in the penetration of double-stranded DNA-binding moieties 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 moieties administered to the eye penetrate into the internal cells and tissues of the eye.

[0036] In a further aspect, a composition is provided that comprises a double-stranded DNA-binding moiety 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 not an antibiotic.

[0037] In some embodiments of a composition suitable for treating a genetic disease, the composition comprises a double-stranded DNA-binding moiety that binds to a repetitive sequence. In some embodiments, the repetitive sequence does not include multiple copies of GAA. In some embodiments, the repetitive sequence includes 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 includes multiple copies of CTG or CAG. In some embodiments, the repetitive sequence includes repeats of at least four nucleotides. In some embodiments, the double-stranded DNA-binding moiety binds to the minor groove of double-stranded DNA.

[0038] In an embodiment of the composition suitable for treating a genetic disease provided herein, 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).

[0039] In the embodiments of the compositions suitable for treating genetic diseases provided herein, the double-stranded DNA binding moiety binds to double-stranded DNA non-covalently.In some embodiments, the double-stranded DNA binding moiety binds to double-stranded DNA using electrostatic force such as hydrogen bond or van der Waals force.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.

[0040] In embodiments of the compositions suitable for treating genetic diseases provided herein, 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, less than about 7.5 kDa, less than about 5 kDa, or less than about 2.5 kDa.

[0041] In another embodiment of the compositions suitable for treating a genetic disease provided herein, the composition comprises a double-stranded DNA-binding moiety, and the double-stranded DNA-binding moiety is a polymer that does not contain nucleotides or amino acids.

[0042] In some embodiments of the composition suitable for treating a genetic disease, 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 connected to the transcriptional modulator-binding moiety by a linker.

[0043] DNA binding part In embodiments of the compositions suitable for treating a genetic disease provided herein, the double-stranded DNA binding moiety is represented by the formula (A-1):

[0044] [ka] or a pharmaceutically acceptable salt thereof, During the ceremony, X 1 , X 2 , X 3 , X 4 , X 5 , X 6 , X 7, and X 8 each independently represents O or NR 2 and Y 1 , Y 2 , Y 3 , Y 4 , Y 5 , Y 6 , Y 7 , and Y 8 are each independently CH or N; 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 , A.A. 1-10 , -Z B -P(O)(OR 1e )2, -Z B -(CH2) p3 -P(O)(OR 1e )2, or -Z B -(CH2) p3 -OP(O)(OR 1e )2, R 1e each independently represents hydrogen, optionally substituted C-C 20 Alkyl, optionally substituted C-C 20 Heteroalkyl, or PEG 1-20 and R 1f each independently represents hydrogen, optionally substituted C-C 20 Alkyl, optionally substituted C-C 20 Heteroalkyl, optionally substituted 5-membered heteroaryl, PEG 1-20 , or AA 1-10 and Each AA is independently a naturally occurring amino acid, Z Bis N or O, p3 is 1 to 10, W 2 is hydrogen, optionally substituted C-C 20 Alkyl, or optionally substituted C-C 20 heteroalkyl; or W 2 Ha-L 1 -ZR 4 and L 1 is alkylene or heteroalkylene, Z is absent, —C(O)—, or —C(═NH)—; R 4 is C1-C6 alkyl, -OR 4b , or -NR 4a R 4b and R 4a is hydrogen, optionally substituted C1-C 20 Alkyl, or optionally substituted C-C 20 is heteroalkyl, R 4b is optionally substituted C1-C 20 Alkyl, optionally substituted C-C 20 Alkenyl, optionally substituted C-C 20 Alkynyl, optionally substituted C-C 20 Aminoalkyl, optionally substituted C-C 20 Haloalkyl, optionally substituted C-C 20 Heteroalkyl, optionally substituted C-C 20 hydroxyalkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted 4-8 membered heterocycloalkyl, optionally substituted phenyl, or optionally substituted 5-10 membered heteroaryl; or R 4a and R 4b together with the nitrogen to which they are attached form a partially or fully unsaturated, optionally substituted 4- to 8-membered heterocycloalkyl; R Wis hydrogen or optionally substituted C-C 20 alkyl, or W 2 and R W together with the nitrogen to which they are attached form a partially or fully unsaturated, optionally substituted 4- to 8-membered heterocycloalkyl; R 2 each independently represents hydrogen, optionally substituted C-C 20 Alkyl, optionally substituted C-C 20 Alkenyl, optionally substituted C-C 20 Alkynyl, optionally substituted C-C 20 Heteroalkyl, optionally substituted C-C 20 Heteroalkenyl, optionally substituted C-C 50 Heteroalkynyl, optionally substituted C-C 20 Hydroxyalkyl, optionally substituted C-C 20 Aminoalkyl, optionally substituted C-C 20 haloalkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted 3-8 membered heterocycloalkyl, or optionally substituted PEG 1-20 and R 3 each independently represents hydrogen, halogen, acetyl, amino, amido, hydroxy, optionally substituted C-C 20 Alkyl, optionally substituted C-C 20 Haloalkyl, optionally substituted C-C 20 alkylamino, or optionally substituted C-C 20 hydroxyalkyl, or The Two R's 3 together with the atom to which they are attached form a C3-C6 cycloalkyl or a 4- to 6-membered heterocycloalkyl; p1 is 3 or 4, n1 and n2 each independently represent 0 or 1; n3 is 0, 1, or 2, m1 is 0, 1, 2, or 3; n0 is 0 or 1, and both n0 and m1 are not 0.

[0045] In some embodiments of Formula (A-1), n0 is 1. In some embodiments of Formula (A-1), n0 is 0.

[0046] In some embodiments of Formula (A-1), n2 is 1. In some embodiments of Formula (A-1), n2 is 0.

[0047] 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.

[0048] In some embodiments of Formula (A-1), p1 is 3. In some embodiments of Formula (A-1), p1 is 4.

[0049] In some embodiments of formula (A-1), X 1 , X 2 , X 3 , X 4 , X 5 , X 6 , X 7 , and X 8 are each independently NR 2 is.

[0050] In some embodiments of Formula (A-1), R 2 each independently represents hydrogen or an optionally substituted C-C 20 In some embodiments of Formula (A-1), R is an alkyl, each of which is optionally substituted with one or more amino, amido, azido, cyano, ester, oxo (=O), urea, optionally substituted aryl, or optionally substituted 5-10 membered heteroaryl. 2 each independently represents an optionally substituted C-C 10In some embodiments of Formula (A-1), R 2 are each independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, pentyl, or hexyl. 2 are each methyl. In some embodiments of Formula (A-1), R 2 are hydrogen atoms.

[0051] In some embodiments of Formula (A-1), R 3 are each independently hydrogen, amino, or amido. In some embodiments of Formula (A-1), R 3 are each independently amino. In some embodiments of Formula (A-1), R 3 are each independently an amide. In some embodiments of Formula (A-1), R 3 are hydrogen atoms.

[0052] In some embodiments of formula (A-1), two R 3 together with the atoms to which they are attached form a C3-C6 cycloalkyl or a 4- to 6-membered heterocycloalkyl. In some embodiments of Formula (A-1), two R 3 together with the atoms to which they are attached form a C3-C6 cycloalkyl. In some embodiments of formula (A-1), two R 3 together with the atom to which they are attached form a 4- to 6-membered heterocycloalkyl.

[0053] In some embodiments of formula (A-1), W 2 Ha-L 1 -ZR 4 is.

[0054] In some embodiments of Formula (A-1), R W is hydrogen.

[0055] In various embodiments of the compositions suitable for treating a genetic disease provided herein, the double-stranded DNA binding moiety is represented by formula (A-2):

[0056] [ka] or a pharmaceutically acceptable salt thereof, During the ceremony, W 1 is hydrogen or -N=C(N(R 1e )2)2 and R 1e are each independently hydrogen or C1-C3 alkyl; Y 1 , Y 2 , Y 3 , Y 4 , Y 5 , Y 6 , Y 7 , and Y 8 are each independently N or CH; L 1 is C1-C 20 Alkylene or C2-C 20 is heteroalkylene, Z is absent, —C(O)—, or —C(═NH)—; R 4 is C1-C6 alkyl, -OR 4b , or -NR 4a R 4b and R 4a is hydrogen, optionally substituted C1-C 20 Alkyl, or optionally substituted C-C 20 is heteroalkyl, R 4b is optionally substituted C1-C 20 Alkyl, optionally substituted C-C 20 Alkenyl, optionally substituted C-C 20 Alkynyl, optionally substituted C-C 20 Aminoalkyl, optionally substituted C-C 20 Haloalkyl, optionally substituted C-C 20Heteroalkyl, optionally substituted C-C 20 hydroxyalkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted 4-8 membered heterocycloalkyl, optionally substituted phenyl, or optionally substituted 5-10 membered heteroaryl; or R 4a and R 4b together with the nitrogen to which they are attached form a partially or fully unsaturated, optionally substituted 4- to 8-membered heterocycloalkyl; R 2a , R 2b , R 2c , R 2d , R 2e , R 2f , R 2g , and R 2h each independently represents hydrogen, optionally substituted C-C 20 Alkyl, optionally substituted C-C 20 Alkenyl, optionally substituted C-C 20 Alkynyl, optionally substituted C-C 20 Heteroalkyl, optionally substituted C-C 20 Heteroalkenyl, optionally substituted C-C 50 Heteroalkynyl, optionally substituted C-C 20 Hydroxyalkyl, optionally substituted C-C 20 Aminoalkyl, optionally substituted C-C 20 haloalkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted 3-8 membered heterocycloalkyl, or optionally substituted PEG 1-20 and each of these is one or more R X is optionally replaced by R 3a and R 3b are each independently hydrogen, halogen, or C1-C 10 Alkyl, -OR 3e , -NR 3c R 3d , or -NHC(O)R 3e and R 3c and R 3d are each independently hydrogen, alkyl, or PEG; R 3e is alkyl, PEG, cycloalkyl, heterocycloalkyl, or phenyl; Or two R's 3a or two R's 3b together with the carbon atoms to which they are attached form a C3-C6 cycloalkyl or a 4- to 6-membered heterocycloalkyl; R X are each independently -CN, -OH, or -OR Xa , -N3, -NR Xa R Xb , -CO(O)R Xc , -C(O)OR Xc , -C(O)NR Xa R Xb , -NHC(O)R Xc , -NHC(O)OR Xc , -OC(O)NR Xa R Xb or an optionally substituted 5-10 membered heteroaryl; R Xa and R Xb are each independently hydrogen, alkyl, or PEG; R Xc is alkyl, PEG, cycloalkyl, heterocycloalkyl, or phenyl, and n1 and m1 each independently represent 0 or 1.

[0057] In some embodiments of formula (A-1) or (A-2), Y 2 , Y 4 , Y 7 , and Y 8 are each independently N. In some embodiments of formula (A-1) or (A-2), Y 1 , Y 3 , and Y 6 are each independently CH.

[0058] In various embodiments of the compositions suitable for treating a genetic disease provided herein, the double-stranded DNA binding moiety is represented by formula (A-3):

[0059] [ka] or a pharmaceutically acceptable salt thereof, During the ceremony, W 1 is hydrogen or -N=C(N(R 1e )2)2 and R 1e are each independently hydrogen or C1-C3 alkyl; Y 5 are CH or N, L 1 is C1-C 20 Alkylene or C2-C 20 is heteroalkylene, Z is absent, —C(O)—, or —C(═NH)—; R 4 is C1-C6 alkyl, -OR 4b , or -NR 4a R 4b and R 4a is hydrogen, optionally substituted C1-C 20 Alkyl, or optionally substituted C-C 20 is heteroalkyl, R 4b is optionally substituted C1-C 20 Alkyl, optionally substituted C-C 20 Alkenyl, optionally substituted C-C 20 Alkynyl, optionally substituted C-C 20 Aminoalkyl, optionally substituted C-C 20 Haloalkyl, optionally substituted C-C 20 Heteroalkyl, optionally substituted C-C 20hydroxyalkyl, optionally substituted C3-C7 cycloalkyl, optionally substituted 4-8 membered heterocycloalkyl, optionally substituted phenyl, or optionally substituted 5-10 membered heteroaryl; or R 4a and R 4b together with the nitrogen to which they are attached form a partially or fully unsaturated, optionally substituted 4- to 8-membered heterocycloalkyl; R 2a , R 2b , R 2c , R 2d , R 2e , R 2f , R 2g , and R 2h each independently represents hydrogen, optionally substituted C-C 20 Alkyl, optionally substituted C-C 20 Alkenyl, optionally substituted C-C 20 Alkynyl, optionally substituted C-C 20 Heteroalkyl, optionally substituted C-C 20 Heteroalkenyl, optionally substituted C-C 50 Heteroalkynyl, optionally substituted C-C 20 Hydroxyalkyl, optionally substituted C-C 20 Aminoalkyl, optionally substituted C-C 20 haloalkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted 3-8 membered heterocycloalkyl, or optionally substituted PEG 1-20 and each of these is one or more R X is optionally replaced by R 3a and R 3b are each independently hydrogen, halogen, or C1-C 10 Alkyl, -OR 3e , -NR 3c R 3d , or -NHC(O)R 3e and R 3c and R 3dare each independently hydrogen, alkyl, or PEG; R 3e is alkyl, PEG, cycloalkyl, heterocycloalkyl, or phenyl; Or two R's 3a or two R's 3b together with the carbon atoms to which they are attached form a C3-C6 cycloalkyl or a 4- to 6-membered heterocycloalkyl; R X are each independently -CN, -OH, or -OR Xa , -N3, -NR Xa R Xb , -CO(O)R Xc , -C(O)OR Xc , -C(O)NR Xa R Xb , -NHC(O)R Xc , -NHC(O)OR Xc , -OC(O)NR Xa R Xb or an optionally substituted 5-10 membered heteroaryl; R Xa and R Xb are each independently hydrogen, alkyl, or PEG; R Xc is alkyl, PEG, cycloalkyl, heterocycloalkyl, or phenyl, and n1 and m1 each independently represent 0 or 1.

[0060] In various embodiments of the compositions suitable for treating a genetic disease provided herein, the double-stranded DNA binding moiety is represented by formula (A-4):

[0061] [ka] or a pharmaceutically acceptable salt thereof, During the ceremony, W 1 is hydrogen or -N=C(N(R 1e )2)2 and R 1eare each independently hydrogen or C1-C3 alkyl; Y 1 , Y 2 , Y 3 , Y 4 , Y 5 , Y 6 , Y 7 , and Y 8 are each independently N or CH; L V is C1-C 20 Alkylene, C2-C 20 Heteroalkylene, or AA 1-10 wherein each AA is independently a naturally occurring amino acid; V is absent, optionally substituted C3-C8 cycloalkyl, optionally substituted 4-8 membered heterocycloalkyl, optionally substituted phenyl, or optionally substituted 5-10 membered heteroaryl; R 2a , R 2b , R 2c , R 2d , R 2e , R 2f , R 2g , and R 2h each independently represents hydrogen, optionally substituted C-C 20 Alkyl, optionally substituted C-C 20 Alkenyl, optionally substituted C-C 20 Alkynyl, optionally substituted C-C 20 Heteroalkyl, optionally substituted C-C 20 Heteroalkenyl, optionally substituted C-C 50 Heteroalkynyl, optionally substituted C-C 20 Hydroxyalkyl, optionally substituted C-C 20 Aminoalkyl, optionally substituted C-C 20 haloalkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted 3-8 membered heterocycloalkyl, or optionally substituted PEG 1-20 and each of these is one or more R Xis optionally replaced by R 3a and R 3b are each independently hydrogen, halogen, or C1-C 10 Alkyl, -OR 3e , -NR 3c R 3d , or -NHC(O)R 3e and R 3c and R 3d are each independently hydrogen, alkyl, or PEG; R 3e is alkyl, PEG, cycloalkyl, heterocycloalkyl, or phenyl; Or two R's 3a or two R's 3b together with the carbon atoms to which they are attached form a C3-C6 cycloalkyl or a 4- to 6-membered heterocycloalkyl; R X are each independently -CN, -OH, or -OR Xa , -N3, -NR Xa R Xb , -CO(O)R Xc , -C(O)OR Xc , -C(O)NR Xa R Xb , -NHC(O)R Xc , -NHC(O)OR Xc , -OC(O)NR Xa R Xb or an optionally substituted 5-10 membered heteroaryl; R Xa and R Xb are each independently hydrogen, alkyl, or PEG; R Xc is alkyl, PEG, cycloalkyl, heterocycloalkyl, or phenyl; n1 and m1 each independently represent 0 or 1; xv is between 0 and 10.

[0062] In some embodiments of formula (A-4), Y 2 , Y 4, Y 7 , and Y 8 are each independently N. In some embodiments of formula (A-4), Y 1 , Y 3 , and Y 6 are each independently CH.

[0063] In some embodiments of formula (A-4), L V is C1-C 20 Alkylene or C2-C 20 In some embodiments of formula (A-4), L is heteroalkylene. V is C1-C 20 In some embodiments of formula (A-4), L is alkylene. V is C1-C 10 In some embodiments of formula (A-4), L is alkylene. V is C2-C 20 In some embodiments of formula (A-4), L is heteroalkylene. V is C2-C 10 In some embodiments of formula (A-4), L is heteroalkylene. V is PEG 1-10 is.

[0064] In some embodiments of Formula (A-4), xv is 0 to 8, 0 to 6, 0 to 4, or 0 to 2. In some embodiments of Formula (A-4), xv is 0 to 8. In some embodiments of Formula (A-4), xv is 0 to 4. In some embodiments of Formula (A-4), xv is 0, 1, 2, or 3.

[0065] In some embodiments, V is an optionally substituted C3-C8 cycloalkyl or an optionally substituted 4-8 membered heterocycloalkyl. In some embodiments, V is an optionally substituted C3-C8 cycloalkyl. In some embodiments, V is an optionally substituted 4-8 membered heterocycloalkyl. In some embodiments, V is an optionally substituted 6 membered heterocycloalkyl. In some embodiments, V is an optionally substituted phenyl or an optionally substituted 5-10 membered heteroaryl. In some embodiments, V is an optionally substituted phenyl. In some embodiments, V is an optionally substituted 5-10 membered heteroaryl. In some embodiments, V is absent.

[0066] In some embodiments of formula (A-4), V is a group represented by formula (C-1):

[0067] [ka] or a pharmaceutically acceptable salt thereof, During the ceremony, B 1 -CR 60 R 60 -, -O-, -NR 60 -, -S(O)-, -S(O)2-, or -S-, or B 1 teeth

[0068] [ka] and R 60 each independently represents hydrogen, optionally substituted C-C 20 Alkyl, optionally substituted C-C 20 Alkenyl, optionally substituted C-C 20 Alkynyl, optionally substituted C-C 20 Heteroalkyl, -C(O)OR 60a , or -C(O)R60a and R 60a is hydrogen, optionally substituted C-C 20 Alkyl, optionally substituted C-C 10 Haloalkyl, optionally substituted PEG 1-20 , optionally substituted C3-C6 cycloalkyl, optionally substituted 3-6 membered heterocycloalkyl, or optionally substituted phenyl; or The Two R's 60 together with the nitrogen atom to which they are attached form an optionally substituted 4- to 8-membered heterocycloalkyl; Ring P is an optionally substituted C3-C6 cycloalkyl, an optionally substituted 4-6 membered heterocycloalkyl, an optionally substituted phenyl, or an optionally substituted 5-10 membered heteroaryl; L b is absent, C1-C4 alkylene, C2-C4 alkenylene, or C2-C4 alkynylene; a1 and a2 are each independently 0, 1, or 2.

[0069] In some embodiments of formula (C-1), B 1 Ha-CR 60 R 60 -, -O-, -NR 60 In some embodiments of formula (C-1), B is -, -S(O)-, -S(O)2-, or -S-. 1 teeth

[0070] [ka] is.

[0071] In some embodiments of Formula (C-1), R 60 each independently represents hydrogen, optionally substituted C-C 20 Alkyl, optionally substituted C-C 20 Alkenyl, optionally substituted C-C 20Alkynyl, or optionally substituted C-C 20 In some embodiments of Formula (C-1), R 60 are each independently -C(O)OR 60a or -C(O)R 60a In some embodiments of formula (C-1), R 60 are hydrogen atoms.

[0072] In some embodiments of formula (C-1), two R 60 taken together with the nitrogen atom to which they are attached form an optionally substituted 4- to 8-membered heterocycloalkyl.

[0073] In some embodiments of formula (A-4), V is a group of formula (C-2):

[0074] [ka] or a pharmaceutically acceptable salt thereof, During the ceremony, Ring P is an optionally substituted C3-C6 cycloalkyl, an optionally substituted 4-6 membered heterocycloalkyl, an optionally substituted phenyl, or an optionally substituted 5-10 membered heteroaryl; L b is absent, C1-C4 alkylene, C2-C4 alkenylene, or C2-C4 alkynylene; B 1’ is CH or N, a1 and a2 are each independently 0, 1, or 2.

[0075] In some embodiments of Formula (C-2), ring P is an optionally substituted C3-C6 cycloalkyl or an optionally substituted 4-6 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 4-6 membered heterocycloalkyl. In some embodiments of Formula (C-2), ring P is an optionally substituted 6 membered heterocycloalkyl.

[0076] 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 independently 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.

[0077] In some embodiments of formula (A-4), V is a group of formula (C-3):

[0078] [ka] or a pharmaceutically acceptable salt thereof, During the ceremony, B 1’ and B 2 are each independently CH or N; B 3 -CR 61 R 61 -, -O-, -S-, -S(O)-, -S(O)2-, or -NR 61 - and R 61 each independently represents hydrogen, optionally substituted C-C 20 Alkyl, optionally substituted C-C 20 Alkenyl, optionally substituted C-C 20 Alkynyl, optionally substituted C-C20 Heteroalkyl, -C(O)OR 61a , or -C(O)R 61a and R 61a is hydrogen, optionally substituted C1-C 20 Alkyl, optionally substituted C-C 10 Haloalkyl, optionally substituted PEG 1-20 , optionally substituted C3-C6 cycloalkyl, optionally substituted 3- to 6-membered heterocycloalkyl, or optionally substituted phenyl; L b is absent, C1-C4 alkylene, C2-C4 alkenylene, or C2-C4 alkynylene.

[0079] 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.

[0080] 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 does not exist.

[0081] In some embodiments of formula (A-4), V is a group of formula (C-4):

[0082] [ka] or a pharmaceutically acceptable salt thereof, During the ceremony, B 2 is CH or N, B 3 -CR 61 R 61 -, -O-, -S-, -S(O)-, -S(O)2-, or -NR 61 - and R 61 each independently represents hydrogen, optionally substituted C-C 20 Alkyl, optionally substituted C-C 20 Alkenyl, optionally substituted C-C 20 Alkynyl, optionally substituted C-C 20 Heteroalkyl, -C(O)OR 61a , or -C(O)R 61a and R 61a is hydrogen, optionally substituted C1-C 20 Alkyl, optionally substituted C-C 10 Haloalkyl, optionally substituted PEG 1-20 , optionally substituted C3-C6 cycloalkyl, optionally substituted 3- to 6-membered heterocycloalkyl, or optionally substituted phenyl; R 62a is hydrogen, optionally substituted C1-C 20 Alkylene, or optionally substituted PEG 1-20 and R 62 are each independently hydrogen or C1-C3 alkyl; s2 is 1, 2, or 3.

[0083] 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.

[0084] In some embodiments of formula (C-3) or (C-4), B 3 Ha-CR 61 R 61 -, -O-, or -NR 61In some embodiments of formula (C-3) or (C-4), B 3 Ha-CR 61 R 61 In some embodiments of formula (C-3) or (C-4), B 3 In some embodiments of formula (C-3) or (C-4), B 3 Ha-NR 61 In some embodiments of formula (C-3) or (C-4), B 3 is -S-, -S(O)-, or -S(O)2-.

[0085] In some embodiments of formula (C-3) or (C-4), R 61 each independently represents hydrogen, optionally substituted C-C 20 Alkyl, optionally substituted C-C 20 Alkenyl, optionally substituted C-C 20 Alkynyl, or optionally substituted C-C 20 In some embodiments of formula (C-3) or (C-4), R 61 are each independently -C(O)OR 61a or -C(O)R 61a In some embodiments of formula (C-3) or (C-4), R 61 are hydrogen atoms.

[0086] In some embodiments of formula (C-3) or (C-4), R 61a is optionally substituted C1-C 20 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 an optionally substituted PEG 1-20 In some embodiments of formula (C-3) or (C-4), R 61a is hydrogen.

[0087] In some embodiments of formula (C-4), R 62 are each independently C1-C3 alkyl. In some embodiments of Formula (C-4), R 62 are hydrogen atoms.

[0088] In some embodiments of formula (C-4), R 62a is an optionally substituted C1-C 20 Alkylene or optionally substituted PEG 1-20 In some embodiments of formula (C-4), R 62a is hydrogen.

[0089] 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.

[0090] In some embodiments of formula (A-2), (A-3), or (A-4), R 2a , R 2b , R 2c , R 2d , R 2e , R 2f , R 2g , and R 2h each independently represents hydrogen, optionally substituted C-C 20 Alkyl, optionally substituted C-C 20 Heteroalkyl, optionally substituted C-C 20 Hydroxyalkyl, optionally substituted C-C 20 Aminoalkyl, optionally substituted C-C 20 haloalkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted 3-8 membered heterocycloalkyl, or optionally substituted PEG 1-20 and each of these is one or more R X In some embodiments of formula (A-2), (A-3), or (A-4), R 2a , R 2b , R 2c, R 2d , R 2e , R 2f , R 2g , and R 2h each independently represents hydrogen, optionally substituted C-C 20 Alkyl, optionally substituted C-C 20 Haloalkyl, optionally substituted C-C 20 heteroalkyl, optionally substituted C-C cycloalkyl, and optionally substituted 4- to 8-membered heterocycloalkyl, each of which may be substituted with one or more R X In some embodiments of formula (A-2), (A-3), or (A-4), R 2a , R 2b , R 2c , R 2d , R 2e , R 2f , R 2g , and R 2h each independently represents hydrogen, optionally substituted C-C 20 Alkyl, optionally substituted C-C 20 haloalkyl, optionally substituted C-C cycloalkyl, and optionally substituted 4- to 8-membered heterocycloalkyl, each of which may be substituted with one or more R X In some embodiments of formula (A-2), (A-3), or (A-4), R 2a , R 2b , R 2c , R 2d , R 2e , R 2f , R 2g , and R 2h each independently represents hydrogen or an optionally substituted C-C 20 alkyl, each of which may be one or more R X In some embodiments of formula (A-2), (A-3), or (A-4), R 2a , R 2b , R 2c , R 2d , R 2e , R 2f , R 2g, and R 2h are each independently an optionally substituted C3-C8 cycloalkyl or an optionally substituted 4-8 membered heterocycloalkyl. In some embodiments of Formula (A-2), (A-3), or (A-4), R 2a , R 2b , R 2c , R 2d , R 2e , R 2f , R 2g , and R 2h are each independently an optionally substituted C-C cycloalkyl. In some embodiments of formula (A-2), (A-3), or (A-4), R 2a , R 2b , R 2c , R 2d , R 2e , R 2f , R 2g , and R 2h are each independently an optionally substituted 4-8 membered heterocycloalkyl. In some embodiments of Formula (A-2), (A-3), or (A-4), R 2a , R 2b , R 2c , R 2d , R 2e , R 2f , R 2g , and R 2h each independently represents an optionally substituted C-C 20 In some embodiments of formula (A-2), (A-3), or (A-4), R 2a , R 2b , R 2c , R 2d , R 2e , R 2f , R 2g , and R 2h each independently represents an optionally substituted C-C 10 In some embodiments of formula (A-2), (A-3), or (A-4), R 2a , R 2b , R 2c , R 2d , R 2e , R2f , R 2g , and R 2h are each 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), R 2a , R 2b , R 2c , R 2d , R 2e , R 2f , R 2g , and R 2h and each is methyl. In some embodiments of formula (A-2), (A-3), or (A-4), R 2a , R 2b , R 2c , R 2d , R 2e , R 2f , R 2g , and R 2h are hydrogen atoms.

[0091] In some embodiments of formula (A-2), (A-3), or (A-4), R 3a and R 3b are each independently hydrogen, -NR 3c R 3d , or -NHC(O)R 3f In some embodiments of formula (A-2), (A-3), or (A-4), R 3a and R 3b are each independently hydrogen or -NR 3c R 3d In some embodiments of formula (A-2), (A-3), or (A-4), R 3a and R 3b are each independently hydrogen or -NH2. In some embodiments of formula (A-2), (A-3), or (A-4), R 3a and R 3b are hydrogen atoms.

[0092] In some embodiments of formula (A-2), (A-3), or (A-4), two R3a together with the carbon atoms to which they are attached form a C3-C6 cycloalkyl. In some embodiments of formula (A-2), (A-3), or (A-4), two R 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 R 3a together with the carbon atom to which they are attached form a cyclopropyl. In some embodiments of formula (A-2), (A-3), or (A-4), two R 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 R 3a together with the carbon atoms to which they are attached form cyclopentyl. In some embodiments of formula (A-2), (A-3), or (A-4), two R 3a together with the carbon atoms to which they are attached form a 4- to 6-membered heterocycloalkyl.

[0093] In some embodiments of formula (A-2), (A-3), or (A-4), two R 3b together with the carbon atoms to which they are attached form a C3-C6 cycloalkyl. In some embodiments of formula (A-2), (A-3), or (A-4), two R 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 R 3b together with the carbon atom to which they are attached form a cyclopropyl. In some embodiments of formula (A-2), (A-3), or (A-4), two R 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 R 3btogether with the carbon atoms to which they are attached form cyclopentyl. In some embodiments of formula (A-2), (A-3), or (A-4), two R 3b together with the carbon atoms to which they are attached form a 4- to 6-membered heterocycloalkyl.

[0094] In some embodiments of formula (A-1), (A-2), or (A-3), L 1 is C1-C 10 Alkylene or C2-C 10 In some embodiments of formula (A-1), (A-2), or (A-3), L is heteroalkylene. 1 is C1-C 10 In some embodiments, L is alkylene. 1 is C2-C 10 In some embodiments of Formula (A-1), (A-2), or (A-3), the heteroalkylene 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 Ha-(CH2CH2-O) y1 - and y1 is an integer ranging from 1 to 10. In some embodiments of formula (A-1), (A-2), or (A-3), the heteroalkylene is -(CH2) x3 N(R a )(CH2) X4 - and R a is hydrogen or optionally substituted C1-C6 alkyl, and x3 and x4 are each independently an integer ranging from 1 to 6.

[0095] In some embodiments of formula (A-1), (A-2), or (A-3), Z is —C(O)— and R 4 Ha-NR 4a R 4bIn some embodiments of formula (A-1), (A-2), or (A-3), Z is absent and R 4 HA-OR 4b In some embodiments of formula (A-1), (A-2), or (A-3), Z is absent and R 4 Ha-NR 4a R 4b In some embodiments of formula (A-1), (A-2), or (A-3), Z is —C(O)— and R 4 is an optionally substituted C1-C6 alkyl. In some embodiments of formula (A-1), (A-2), or (A-3), Z is absent and R 4 is an optionally substituted C1-C6 alkyl.

[0096] In some embodiments of formula (A-1), (A-2), or (A-3), R 4a is hydrogen, optionally substituted C-C 20 Alkyl, or optionally substituted C-C 20 In some embodiments of formula (A-1), (A-2), or (A-3), R 4a is an optionally substituted C1-C 20 Alkyl or optionally substituted C-C 20 In some embodiments of formula (A-1), (A-2), or (A-3), R 4a is optionally substituted C1-C 20 In some embodiments of formula (A-1), (A-2), or (A-3), R 4a is optionally substituted C1-C 20 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 an optionally substituted PEG 1-20 In some embodiments of formula (A-1), (A-2), or (A-3), R4a is hydrogen.

[0097] In some embodiments of formula (A-1), (A-2), or (A-3), R 4b is hydrogen, optionally substituted C-C 20 Alkyl, or optionally substituted C-C 20 In some embodiments of formula (A-1), (A-2), or (A-3), R 4b is an optionally substituted C1-C 20 Alkyl or optionally substituted C-C 20 In some embodiments of formula (A-1), (A-2), or (A-3), R 4b is optionally substituted C1-C 20 In some embodiments of formula (A-1), (A-2), or (A-3), R 4b is optionally substituted C1-C 20 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 is an optionally substituted PEG 1-20 In some embodiments of formula (A-1), (A-2), or (A-3), R 4b is hydrogen.

[0098] In some embodiments of formula (A-1), (A-2), or (A-3), R 4b is an optionally substituted C3-C8 cycloalkyl, an optionally substituted 4-8 membered heterocycloalkyl, an optionally substituted phenyl, or an optionally substituted 5-10 membered heteroaryl. In some embodiments of Formula (A-1), (A-2), or (A-3), R 4bis an optionally substituted C3-C8 cycloalkyl or an optionally substituted 4-8 membered heterocycloalkyl. In some embodiments of Formula (A-1), (A-2), or (A-3), R 4b is an optionally substituted C-C cycloalkyl. In some embodiments of formula (A-1), (A-2), or (A-3), R 4b is an optionally substituted 4-6 membered heterocycloalkyl.

[0099] 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 4-8 membered heterocycloalkyl that is partially 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.

[0100] In some embodiments of formula (A-2), (A-3), or (A-4), R x are each independently -CN, -OH, or -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), R x are each independently -CN, -OH, or -OR Xa , -N3, or -NR Xa R XbIn some embodiments of formula (A-2), (A-3), or (A-4), R x are each independently -CO(O)R Xc , -C(O)OR Xc , -C(O)NR Xa R Xb , or -NHC(O)R Xc is.

[0101] In some embodiments of formula (A-2), (A-3), or (A-4), R Xa and R Xb 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 Xa and R Xb are each independently, C1-C 20 In some embodiments of formula (A-2), (A-3), or (A-4), R Xa and R Xb are each independently PEG 1-20 In some embodiments of formula (A-2), (A-3), or (A-4), R Xa and R Xb are hydrogen atoms.

[0102] 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, 4-6 membered heterocycloalkyl, 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 In some embodiments of formula (A-2), (A-3), or (A-4), R Xcis PEG 1-20 is.

[0103] 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 In some embodiments of formula (A-2), (A-3), or (A-4), R 3c and R 3d are each independently hydrogen.

[0104] 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, 4-6 membered heterocycloalkyl, 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 is.

[0105] In some embodiments of formula (A-1), (A-2), (A-3), or (A-4), Y 5 are each independently N. In some embodiments of formula (A-1), (A-2), (A-3), or (A-4), Y 5 are each independently CH.

[0106] In some embodiments of formula (A-1), (A-2), (A-3), or (A-4), W 1 is -N=C(N(R 1e )2)2 and R 1eare each independently hydrogen or C1-C3 alkyl. In some embodiments of formula (A-1), (A-2), (A-3), or (A-4), W 1 is hydrogen.

[0107] 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.

[0108] 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.

[0109] In the embodiments of the compositions suitable for treating genetic diseases provided herein, the double-stranded DNA binding moiety is optionally linked to the second end comprising the moiety that regulates gene expression. In some embodiments, the double-stranded DNA binding moiety is linked to the second end comprising the moiety that regulates gene expression by a linker.

[0110] In some embodiments, the second end comprising the gene expression-modulating moiety is conjugated via a pyrrole on the double-stranded DNA-binding moiety. In some embodiments, the second end comprising the gene expression-modulating moiety is conjugated via an imidazole on the double-stranded DNA-binding moiety. In some embodiments, the second end comprising the gene expression-modulating moiety is conjugated via a β-alanine on the double-stranded DNA-binding moiety. In some embodiments, the second end comprising the gene expression-modulating moiety is conjugated via the C-terminus of the double-stranded DNA-binding moiety. In some embodiments, the second end comprising the gene expression-modulating moiety is conjugated via the N-terminus of the double-stranded DNA-binding moiety. In some embodiments, the second end comprising the gene expression-modulating moiety is conjugated via an amide on the double-stranded DNA-binding moiety.

[0111] In an embodiment of a composition suitable for treating a genetic disease provided herein, the double-stranded DNA-binding moiety of formula (A-1), (A-2), or (A-3) is optionally linked by an oligomeric linker to a second end comprising a moiety that regulates gene expression. In an embodiment of a composition suitable for treating a genetic disease provided herein, the double-stranded DNA-binding moiety of formula (A-1) is optionally linked to a second end comprising a moiety that regulates gene expression. In an embodiment of a composition suitable for treating a genetic disease provided herein, the double-stranded DNA-binding moiety of formula (A-2) is optionally linked to a second end comprising a moiety that regulates gene expression. In an embodiment of a composition suitable for treating a genetic disease provided herein, the double-stranded DNA-binding moiety of formula (A-3) is optionally linked to a second end comprising a moiety that regulates gene expression.

[0112] In some embodiments, the second end containing the gene expression modulating moiety is 1 In some embodiments, the second end containing the gene expression modulating moiety is conjugated with W 2 In some embodiments, the second end containing the gene expression-regulating moiety is conjugated with R2 In some embodiments, the second end containing the gene expression-regulating 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 containing the gene expression-regulating moiety is conjugated at one of R 3 In some embodiments, the second end containing the gene expression-regulating moiety is conjugated at one of R 3a In some embodiments, the second end containing the gene expression-regulating moiety is conjugated at one of R 3b In some embodiments, the second end containing the gene expression-regulating moiety is conjugated at one of R 4 In some embodiments, the second end containing the gene expression-regulating moiety is conjugated with R 4a or R 4b It is conjugated with

[0113] The part that regulates gene expression In some embodiments, the second end comprises a moiety that regulates gene expression.

[0114] In some embodiments, the second end comprises a bromodomain binding moiety.

[0115] In some embodiments, the second end comprises a bromodomain and a moiety capable of binding to a BET family member.

[0116] 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.

[0117] In some embodiments, the protein binding moiety binds to CBP / p300, PCAF (P300 / CBP associated factor), CECR2 (cat eye syndrome chromosomal region candidate 2), BRPF (bromodomain and PHD finger containing protein), ATAD2 / ATAD2B (chromatin remodeling protein), TRIM24 (tripartite motif containing 24), BAZ2 (bromodomain adjacent to zinc finger), TAF1 (TBP associated factor), BRD7 / 9, BPTF (bromodomain PHD finger transcription factor), SMARCA2 / 4, or PBRM1.

[0118] In some embodiments, the regulatory molecule is CBP / p300.

[0119] In some embodiments, the regulatory molecule is PCAF (P300 / CBP associated factor).

[0120] In some embodiments, the regulatory molecule is CECR2 (Cat Eye Syndrome Chromosomal Region Candidate 2).

[0121] In some embodiments, the regulatory molecule is BRPF (bromodomain and PHD finger-containing protein).

[0122] In some embodiments, the regulatory molecule is an ATAD2 or ATAD2B chromatin remodeling protein.

[0123] In some embodiments, the regulatory molecule is BAZ2 (zinc finger adjacent bromodomain).

[0124] In some embodiments, the regulatory molecule is TAF1 (TBP-associated factor).

[0125] In some embodiments, the regulatory molecule is TRIM24 (tri-element motif-containing 24).

[0126] In some embodiments, the regulatory molecule is BRD7 / 9.

[0127] In some embodiments, the regulatory molecule is BPTF (bromodomain PHD finger transcription factor).

[0128] In some embodiments, the regulatory molecule is SMARCA2 / 4.

[0129] In some embodiments, the regulatory molecule is PBRM1.

[0130] In some embodiments, the regulatory molecule regulates histone rearrangement.

[0131] In some embodiments, the regulatory molecule modulates glycosylation, phosphorylation, alkylation, or acylation of histones.

[0132] In some embodiments, the regulatory molecule is a transcription factor.

[0133] In some embodiments, the regulatory molecule is an RNA polymerase.

[0134] In some embodiments, the regulatory molecule is a moiety that regulates the activity of an RNA polymerase.

[0135] In some embodiments, the recruiting moiety binds to a regulatory molecule but does not inhibit the activity of the regulatory molecule. In some embodiments, the recruiting moiety binds to a regulatory molecule and inhibits the activity of the regulatory molecule. In some embodiments, the recruiting moiety binds to a regulatory molecule and increases the activity of the regulatory molecule.

[0136] In some embodiments, the recruiting moiety binds to an active site of the regulatory molecule. In certain embodiments, the recruiting moiety binds to a regulatory site of the regulatory molecule.

[0137] In some embodiments, the second end is represented by formula (2-A):

[0138] [ka] or a pharmaceutically acceptable salt thereof, During the ceremony, Ring A is an optionally substituted aryl or an optionally substituted 5-6 membered heteroaryl; Ring B is absent or is an optionally substituted 6-membered monocyclic aryl or heteroaryl; D is C or N, E is O or N, Y A is -NH- or -O-, R 5 is hydrogen or C1-C6 alkyl, R 6 is selected from hydrogen, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 haloalkyl, and optionally substituted C1-C6 hydroxyalkyl; R 7 is hydrogen, halogen, -NO2, -CN, optionally substituted aryl, optionally substituted C1-C 20 Alkyl, optionally substituted C-C 20 selected from heteroalkyl, optionally substituted C-C haloalkyl, and optionally substituted C-C hydroxyalkyl; Or, R 7 Ha-NR 7A R 7B and R 7A and R 7B each independently represents hydrogen, optionally substituted C-C 20Alkyl, or optionally substituted C-C 20 is heteroalkyl, x1 is an integer from 1 to 6.

[0139] In some embodiments, D is N and E is N. In some embodiments, D is C and E is O.

[0140] In some embodiments, the second end is represented by formula (2-B):

[0141] [ka] or a pharmaceutically acceptable salt thereof, During the ceremony, Ring A is an optionally substituted aryl or an optionally substituted 5-6 membered heteroaryl; Ring B is absent or is an optionally substituted 6-membered monocyclic aryl or heteroaryl; Y A is -NH- or -O-, R 5 is hydrogen or C1-C6 alkyl, R 6 is selected from hydrogen, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 haloalkyl, and optionally substituted C1-C6 hydroxyalkyl; R 7 is hydrogen, halogen, -NO2, -CN, optionally substituted aryl, optionally substituted C1-C 20 Alkyl, optionally substituted C-C 20 selected from heteroalkyl, optionally substituted C-C haloalkyl, and optionally substituted C-C hydroxyalkyl; Or, R 7 Ha-NR 7A R 7B and R 7A and R 7B each independently represents hydrogen, optionally substituted C-C20 Alkyl, or optionally substituted C-C 20 is heteroalkyl, x1 is an integer from 1 to 6.

[0142] In some embodiments, ring A is an optionally substituted aryl ring. In some embodiments, ring A is an optionally substituted phenyl. In some embodiments, ring A is an optionally substituted 5-membered heteroaryl. In some embodiments, ring A is an optionally substituted oxazolyl, an optionally substituted furanyl, or an optionally substituted thiophenyl.

[0143] In some embodiments, the second end has the formula (2-C):

[0144] [ka] or a pharmaceutically acceptable salt thereof, During the ceremony, R 8 and R 9 are each independently selected from hydrogen, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 haloalkyl, or optionally substituted C1-C6 hydroxyalkyl.

[0145] In some embodiments, R 8 and R 9 are each independently selected from optionally substituted C-C alkyl, C-C haloalkyl, or C-C hydroxyalkyl. 8 and R 9 are each independently selected from optionally substituted C1-C6 alkyl. In some embodiments, R 8 and R 9 are each independently methyl, ethyl, or propyl. In some embodiments, R 8 and R 9are each independently methyl. In some embodiments, R 8 and R 9 are each independently ethyl. In some embodiments, R 8 and R 9 are each independently propyl.

[0146] In some embodiments, the second end has formula (2-D):

[0147] [ka] or a pharmaceutically acceptable salt thereof, During the ceremony, R 10 is selected from hydrogen, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 haloalkyl, or optionally substituted C1-C6 hydroxyalkyl.

[0148] 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.

[0149] In some embodiments, R 7 is selected from hydrogen, halogen, optionally substituted C-C alkyl, C-C haloalkyl, or C-C hydroxyalkyl. 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.

[0150] In some embodiments, R 7 Ha-NR 7A R 7B and R 7A and R 7B are each independently hydrogen or optionally substituted C1-C6 alkyl.

[0151] In some embodiments, R 10 is selected from optionally substituted C1-C6 alkyl, optionally substituted C1-C6 haloalkyl, or optionally substituted C1-C6 hydroxyalkyl. In some embodiments, R 10 is selected from optionally substituted C1-C6 alkyl. In some embodiments, R 10 is methyl, ethyl, or propyl. In some embodiments, R 10 is methyl. In some embodiments, R 10 is optionally replaced by C 1-6 In some embodiments, R 10 is -OMe.

[0152] In some embodiments, R 6 is selected from optionally substituted C1-C6 alkyl, optionally substituted C1-C6 haloalkyl, or optionally substituted C1-C6 hydroxyalkyl. In some embodiments, R 6 is optionally substituted C1-C6 alkyl. 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.

[0153] In some embodiments, Y A In some embodiments, Y A is —O—. In some embodiments, Y A is NH and x1 is 1.

[0154] 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.

[0155] In some embodiments, Ring B is an optionally substituted 6-membered monocyclic aryl or heteroaryl, each of which is optionally substituted with alkyl, amino, halogen, hydroxy, hydroxyalkyl, or PEG. In some embodiments, Ring B is phenyl. In some embodiments, Ring B is a 6-membered monocyclic heteroaryl. In some embodiments, Ring B is pyridine or pyrimidine. In some embodiments, Ring B is absent.

[0156] In some embodiments, the second end is a group represented by formula (2-E), (2-F), or (2-G):

[0157] [ka] or a pharmaceutically acceptable salt thereof.

[0158] In some embodiments, the second end is represented by formula (3-A):

[0159] [ka] or a pharmaceutically acceptable salt thereof, During the ceremony, Y B is -CH2NH-, -CHO-, -NH-, or -O-, R 11A and R11B are each independently hydrogen or optionally substituted C1-C6 alkyl; R 12 is hydrogen, halogen, —OH, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 haloalkyl, or optionally substituted C1-C6 hydroxyalkyl; R 14 and R 15 are each independently hydrogen, halogen, —CN, —NO, optionally substituted C-C alkyl, optionally substituted C-C haloalkyl, or optionally substituted C-C hydroxyalkyl; Alternatively, R 14 Ha-NR A R B and R 16 is an optionally substituted C1-C6 alkyl, an optionally substituted C1-C6 heteroalkyl, an optionally substituted C2-C6 alkenyl, an optionally substituted C2-C6 alkynyl, a C1-C6 hydroxyalkyl, -SO2R A , or -NHSO2R A and R YA is hydrogen, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 haloalkyl, or optionally substituted 5-6 membered monocyclic aryl or heteroaryl; R A and R B are each 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 4-6 membered heterocycloalkyl, or optionally substituted 4-6 membered heteroalkyl; y1 is 1 to 3, The bond to the linker is R 14 or R YA It is in one of the following.

[0160] In some embodiments, the second end is represented by formula (3-B):

[0161] [ka] or a pharmaceutically acceptable salt thereof, During the ceremony, Ring C is absent or an optionally substituted 5- to 6-membered monocyclic aryl or heteroaryl, or a 4- to 8-membered heterocycle; Y B is -NH-, -CHNH-, -CHO-, or -O-, R 11A and R 11B are each independently hydrogen or optionally substituted C1-C6 alkyl; R 12 is hydrogen, optionally substituted C1-C6 alkyl, C(O)R A , or C(O)NR A R B and R A and R B are each independently hydrogen, optionally substituted C-C alkyl, or optionally substituted C-C heteroalkyl; R 13 is hydrogen, substituted aryl, substituted heteroaryl, or substituted oxydibenzene; y2 is an integer from 0 to 2.

[0162] In some embodiments, y2 is 0. In some embodiments, y2 is 1. In some embodiments, y2 is 2.

[0163] In some embodiments, R 13 is substituted aryl or substituted heteroaryl. In some embodiments, R 13 is hydrogen.

[0164] In some embodiments, R 13 is a substituted oxydibenzene.

[0165] In some embodiments, R 13 teeth,

[0166] [ka] wherein: R 14 and R 15 are each independently hydrogen, halogen, —CN, —NO, optionally substituted C-C alkyl, optionally substituted C-C haloalkyl, or optionally substituted C-C hydroxyalkyl; Alternatively, R 14 Ha-NR A R B and R 16 is a substituted C1-C6 alkyl, an optionally substituted C1-C6 heteroalkyl, an optionally substituted C2-C6 alkenyl, an optionally substituted C2-C6 alkynyl, an optionally substituted C1-C6 hydroxyalkyl, -SO2R A , or -NHSO2R A and R A and R B are each 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 heterocycloalkyl, or optionally substituted 5- to 6-membered heteroaryl; y1 is 1 to 3.

[0167] In some embodiments, the second end is represented by formula (3-C):

[0168] [ka] or a pharmaceutically acceptable salt thereof, During the ceremony, Ring C is absent or an optionally substituted 5- to 6-membered monocyclic aryl or heteroaryl, or a 4- to 8-membered heterocycle; Y B is -CH2NH-, -CHO-, -NH-, or -O-, R 11A and R 11B are each independently hydrogen or optionally substituted C1-C6 alkyl; R 12 is hydrogen, halogen, —OH, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 haloalkyl, or optionally substituted C1-C6 hydroxyalkyl; R 14 and R 15 are each independently hydrogen, halogen, —CN, —NO, optionally substituted C-C alkyl, optionally substituted C-C haloalkyl, or optionally substituted C-C hydroxyalkyl; Alternatively, R 14 Ha-NR A R B and R 16 is a substituted C1-C6 alkyl, an optionally substituted C1-C6 heteroalkyl, an optionally substituted C2-C6 alkenyl, an optionally substituted C2-C6 alkynyl, an optionally substituted C1-C6 hydroxyalkyl, -SO2R A , or -NHSO2R A and R A and R B are each 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 heterocycloalkyl, or optionally substituted 5- to 6-membered heteroaryl; y1 is an integer of 1 to 3.

[0169] In some embodiments, Y B In some embodiments, Y B is —CH 2 NH—. In some embodiments, Y B is —CH2O—. In some embodiments, Y B is -O-.

[0170] In some embodiments, Ring C is an optionally substituted 5- or 6-membered monocyclic aryl or heteroaryl, each of which 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, Ring C is oxazole or oxadiazole. In some embodiments, Ring C is thiazole or thiadiazole. In some embodiments, Ring C is absent.

[0171] In some embodiments, the second end has formula (3-D):

[0172] [ka] or a pharmaceutically acceptable salt thereof, During the ceremony, R 11Aand R 11B are each independently hydrogen or optionally substituted C1-C6 alkyl; R 12 is hydrogen, halogen, —OH, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 haloalkyl, optionally substituted C1-C6 hydroxyalkyl, C(O)R A , or C(O)NR A R B and R 15 are each independently hydrogen, halogen, —CN, —NO, optionally substituted C-C alkyl, optionally substituted C-C haloalkyl, or optionally substituted C-C hydroxyalkyl; R 16 is an optionally substituted C1-C6 alkyl, an optionally substituted C1-C6 heteroalkyl, an optionally substituted C2-C6 alkenyl, an optionally substituted C2-C6 alkynyl, an optionally substituted C 1-6 Hydroxyalkyl, -SO2R A , or -NHSO2R A and R A and R B are each 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 heterocycloalkyl, or optionally substituted 5- to 6-membered heteroaryl; y1 is an integer of 1 to 3.

[0173] In some embodiments, R 11A and R 11B are each independently an optionally substituted C1-C6 alkyl. In some embodiments, R 11A and R 11Bare each independently methyl, ethyl, propyl, or tert-butyl. 11A and R 11B are each independently methyl. In some embodiments, R 11A and R 11B are each independently hydrogen.

[0174] In some embodiments, R 11A is C1-C6 alkyl optionally substituted with haloalkyl or phosphorus hydroxide. In some embodiments, R 11A is C1-C6 alkyl substituted with -OP(O)(OH). 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.

[0175] In some embodiments, R 12 is an 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 and R A and R B are each independently hydrogen or optionally substituted C1-C6 alkyl.

[0176] In some embodiments, R 14 and R 15 are each independently hydrogen, —CN, or —NO. In some embodiments, R 14 and R 15are each independently halogen or optionally substituted C-C alkyl. In some embodiments, R 14 and R 15 are each independently Br, Cl, F, methyl, or ethyl. In some embodiments, R 14 and R 15 are each independently F or methyl.

[0177] In some embodiments, R 16 is an optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, or optionally substituted C1-C6 hydroxyalkyl, each of which is optionally substituted with amido, alkyl, alkynyl, azido, amino, halogen, haloalkyl, hydroxy, nitro, oxo (=O), phosphorus hydroxide, or PEG. In some embodiments, R 16 is optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, or optionally substituted C1-C6 hydroxyalkyl. In some embodiments, R 16 is C1-C6 alkyl or C1-C6 heteroalkyl, each of which is optionally substituted with -CN, -NH2, -N3, -OH, CF3, or -OP(O)(OH). In some embodiments, R 16 Ha-SO2R A and R A is C1-C6 alkyl. In some embodiments, R 16 is -SO2Et. In some embodiments, R 16 is -SOMe. In some embodiments, R 16 Ha-NHSO2R A and R A is C1-C6 alkyl. In some embodiments, R 16 is -NHSO2Et. In some embodiments, R 16 is -NHSO2Me.

[0178] In some embodiments, y1 is 1. In some embodiments, y1 is 2. In some embodiments, y1 is 3.

[0179] In some embodiments, the second end is represented by formula (3-E) or formula (3-F):

[0180] [ka] or a pharmaceutically acceptable salt thereof.

[0181] In some embodiments, the second end is represented by formula (3-G) or formula (3-H):

[0182] [ka] or a pharmaceutically acceptable salt thereof.

[0183] In some embodiments, the second end is represented by formula (4-A):

[0184] [ka] or a pharmaceutically acceptable salt thereof, During the ceremony, Ring D is absent, phenyl or 5- to 6-membered heteroaryl; X 9 and X 10 are each independently C or N, and X 9 or X 10 One of them is N, L 2 is absent or is an optionally substituted alkylene, -O-, or -NR D - and R D is hydrogen or optionally substituted C1-C3 alkyl; R 18is an optionally substituted 5-6 membered heteroaryl; R 19 is an optionally substituted C3-C8 cycloalkyl or an optionally substituted 4-7 membered heteroaryl; R 20 are each independently hydrogen, halogen, —CN, —NO, optionally substituted C-C alkyl, optionally substituted C-C haloalkyl, or optionally substituted C-C hydroxyalkyl; X3 is an integer from 1 to 3, y4 is an integer from 1 to 4, The bond to the linker is R 19 or R 20 It is in one of the following.

[0185] In some embodiments, the bond to the linker is R 19 In some embodiments, the bond to the linker is 20 It is one of the.

[0186] In some embodiments, the second end is represented by formula (4-B):

[0187] [ka] or a pharmaceutically acceptable salt thereof, During the ceremony, Ring D is absent, optionally substituted phenyl, or optionally substituted 5-6 membered heteroaryl; X 9 and X 10 are each independently C or N, and X 9 or X 10 One of them is N, L 2 is absent or is an optionally substituted alkylene, -O-, or -NR D - and R D is hydrogen or optionally substituted C1-C3 alkyl; R 18is an optionally substituted 5-6 membered heteroaryl; R 19 is an optionally substituted C3-C8 cycloalkyl or an optionally substituted 4-7 membered heteroaryl; x3 is an integer from 1 to 3.

[0188] In some embodiments, X 9 is N and X 10 is C. In some embodiments, X 9 is C and X 10 is N.

[0189] In some embodiments, the second end is represented by formula (4-C):

[0190] [ka] or a pharmaceutically acceptable salt thereof, During the ceremony, Ring D is absent, optionally substituted phenyl, or optionally substituted 5-6 membered heteroaryl; L 2 is absent or is an optionally substituted alkylene, -O-, or -NR D - and R D is hydrogen or optionally substituted C1-C3 alkyl; R 18 is an optionally substituted 5-6 membered heteroaryl; R 19 is an optionally substituted C3-C8 cycloalkyl or an optionally substituted 4-7 membered heteroaryl; x3 is an integer from 1 to 3.

[0191] In some embodiments, Ring D is an optionally substituted monocyclic 6-membered aryl or 5-6-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.

[0192] In some embodiments, R 19 is an optionally substituted C-C cycloalkyl. In some embodiments, R 19 is an optionally substituted 4-7 membered heteroaryl.

[0193] In some embodiments, the second end is represented by formula (4-D):

[0194] [ka] or a pharmaceutically acceptable salt thereof, During the ceremony, L 2 is an optionally substituted alkylene, -O-, or -NR D - and R D is hydrogen or optionally substituted C1-C3 alkyl; R 18 is an optionally substituted 5-6 membered heteroaryl; R 20 is hydrogen, halogen, —CN, —NO, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 haloalkyl, or optionally substituted C1-C6 hydroxyalkyl; X3 is an integer from 1 to 3, y4 is an integer from 1 to 4.

[0195] In some embodiments, L 2 is an optionally substituted alkylene. In some embodiments, L 2 is a C2-C4 alkylene optionally substituted with one or more C1-C3 alkyl.2 is absent. In some embodiments, L 2 Ha-NR D In some embodiments, L 2 is -NH-.

[0196] 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.

[0197] In some embodiments, R 20 is halogen, —CN, —NO 2 , optionally substituted C 1 -C 6 alkyl, optionally substituted C 1 -C 6 haloalkyl, or optionally substituted C 1 -C 6 hydroxyalkyl.

[0198] In some embodiments, x3 is 1. In some embodiments, x3 is 2. In some embodiments, x3 is 3.

[0199] 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.

[0200] In some embodiments, the second end is a group represented by formula (4-E), (4-F), or (4-G):

[0201] [ka] or a pharmaceutically acceptable salt thereof.

[0202] In some embodiments, the second end is represented by formula (5-A):

[0203] [ka] or a pharmaceutically acceptable salt thereof, During the ceremony, Ring E is absent or is an optionally substituted phenyl or an optionally substituted 5- to 6-membered heteroaryl; X 11 is CH or N, L 3 Ha-NR E -or-CR E R E - and R E are each independently hydrogen or optionally substituted C1-C3 alkyl; R 21 is C1-C6 alkyl or C3-C6 cycloalkyl, R 22 is halogen, CN, NO2, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 haloalkyl, or optionally substituted C1-C6 hydroxyalkyl.

[0204] 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 5-6 membered heteroaryl. In some embodiments, ring E is a 5 membered heteroaryl. In some embodiments, ring E is a 6 membered heteroaryl.

[0205] In some embodiments, X 11 is CH and L 3 Ha-NR E In some embodiments, X 11 is N and L 3 Ha-CR E R E -It is.

[0206] In some embodiments, R 21 is C1-C6 alkyl. In some embodiments, R 21 is methyl.

[0207] In some embodiments, R 22 is halogen, optionally substituted C-C alkyl, optionally substituted C-C haloalkyl, or optionally substituted C-C hydroxyalkyl. In some embodiments, R 22 is CN, F, Cl, Br, or methyl.

[0208] In some embodiments, the second end is represented by formula (5-B):

[0209] [ka] or a pharmaceutically acceptable salt thereof.

[0210] In some embodiments, the second end is represented by formula (6-A):

[0211] [ka] or a pharmaceutically acceptable salt thereof, During the ceremony, Ring G is an optionally substituted C3-C6 cycloalkyl or an optionally substituted 4-6 membered heterocycloalkyl; L 6 is -O-(optionally substituted alkylene); R 28 is an optionally substituted 5-6 membered heteroaryl; R 29 is an optionally substituted C1-C6 alkyl (C6-C 10 aryl) or optionally substituted C1-C6 alkyl (6-10 membered heteroaryl); R 30is an optionally substituted C1-C6 alkyl, an optionally substituted C1-C6 haloalkyl, or an optionally substituted C1-C6 hydroxyalkyl.

[0212] In some embodiments, the second end is represented by formula (7-A):

[0213] [ka] or a pharmaceutically acceptable salt thereof, During the ceremony, A 3 is -O-, -NH-, or -CH2-, Z 2 is CH or N, W is O or S; R 31 are each independently hydrogen, halogen, -OH, -CN, -NO2, -NH2, optionally substituted C1-C 10 Alkyl, optionally substituted C-C 10 Haloalkyl, optionally substituted C-C 10 Hydroxyalkyl, optionally substituted C-C 10 Alkenyl, optionally substituted C-C 10 alkynyl, optionally substituted C3-C8 cycloalkyl, or optionally substituted 3- to 8-membered heterocycloalkyl; Or two R's 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; R 32 is hydrogen or optionally substituted C1-C 10 is alkyl, R 33 is hydrogen, halogen, -OH, -CN, -NO2, -NH2, optionally substituted C1-C 10 Alkyl, optionally substituted C-C 10 Haloalkyl, or optionally substituted C-C10 is hydroxyalkyl, q6 is 0 to 4.

[0214] In some embodiments, the second end is represented by formula (7-B):

[0215] [ka] or a pharmaceutically acceptable salt thereof, During the ceremony, Ring F is an optionally substituted 5-6 membered heteroaryl; A 3 is -O-, -NH-, or -CH2-, Z 3 is CH or N, W is O or S; R 31 are hydrogen, halogen, -OH, -CN, -NO2, -NH2, optionally substituted C1-C 10 Alkyl, optionally substituted C-C 10 Haloalkyl, optionally substituted C-C 10 Hydroxyalkyl, optionally substituted C-C 10 Alkenyl, optionally substituted C-C 10 alkynyl, optionally substituted C3-C8-cycloalkyl, or optionally substituted 3- to 8-membered heterocycloalkyl, Or two R's 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; R 32 is hydrogen or optionally substituted C1-C 10 is alkyl, q6 is 1 to 4.

[0216] In some embodiments, A 3 is —O—. In some embodiments, A 3is -NH-. In some embodiments, A 3 is -CH2-.

[0217] In some embodiments, Z 2 is CH. In some embodiments, Z 2 is N.

[0218] In some embodiments, Z 3 is CH. In some embodiments, Z 3 is N.

[0219] In some embodiments, W is O. In some embodiments, W is S.

[0220] In some embodiments, ring F is an optionally substituted 5-membered heteroaryl. In some embodiments, ring F is an optionally substituted 6-membered heteroaryl.

[0221] In some embodiments, R 31 each independently represents an optionally substituted C-C 10 Alkyl, optionally substituted C-C 10 Haloalkyl, or optionally substituted C-C 10 In some embodiments, R 31 are each independently an optionally substituted C-C-cycloalkyl or an optionally substituted 3- to 8-membered heterocycloalkyl. 31 are each independently hydrogen, halogen, —OH, —CN, —NO, or —NH. In some embodiments, R 31 are hydrogen atoms.

[0222] In some embodiments, R 32 is an optionally substituted C1-C 10 In some embodiments, R 32is methyl. In some embodiments, R 32 is hydrogen.

[0223] In some embodiments, R 33 is hydrogen, halogen, —OH, —CN, —NO, or —NH. In some embodiments, R 33 is an optionally substituted C1-C 10 Alkyl, optionally substituted C-C 10 Haloalkyl, or optionally substituted C-C 10 It is a hydroxyalkyl.

[0224] In some embodiments, the second end is represented by formula (7-C):

[0225] [ka] or a pharmaceutically acceptable salt thereof.

[0226] In some embodiments, the second end is represented by formula (7-D):

[0227] [ka] or a pharmaceutically acceptable salt thereof.

[0228] In some embodiments, the second end is represented by formula (8-A):

[0229] [ka] or a pharmaceutically acceptable salt thereof, During the ceremony, Ring H is optionally substituted phenyl or optionally substituted 6-membered heteroaryl; or Ring H is

[0230] [ka] and Z A is absent or is an optionally substituted phenylformamide; X 12 is CH or N, R 34 is an optionally substituted phenyl or an optionally substituted 6-membered heteroaryl; R 34A is hydrogen, halogen, or optionally substituted C1-C3 alkyl; R 35 are independently halogen, optionally substituted C-C 10 alkyl, or optionally substituted 5-6 membered heteroaryl; The bond to the linker is R 35 , Z A , or in ring H.

[0231] 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

[0232] [ka] is.

[0233] 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.

[0234] In some embodiments, X 12 is CH. In some embodiments, X 12 is N.

[0235] In some embodiments, R 34is optionally substituted phenyl. In some embodiments, R 34 is an optionally substituted 6-membered heteroaryl.

[0236] In some embodiments, R 34A is hydrogen or halogen. In some embodiments, R 34A is an optionally substituted C1-C3 alkyl. In some embodiments, R 34A is methyl.

[0237] In some embodiments, formula (8-A) is R 35 In some embodiments, formula (8-A) is a bond to a linker at Z A In some embodiments, Formula (8-A) is attached to the linker at ring H.

[0238] In some embodiments, the second end is represented by formula (8-B) or formula (8-C):

[0239] [ka] or a pharmaceutically acceptable salt thereof.

[0240] In some embodiments, the second end is represented by formula (8-D):

[0241] [ka] or a pharmaceutically acceptable salt thereof.

[0242] In some embodiments, the second end is represented by formula (9-A):

[0243] [ka] or a pharmaceutically acceptable salt thereof.

[0244] In some embodiments, the second end is represented by formula (10-A) or formula (10-B):

[0245] [ka] or a pharmaceutically acceptable salt thereof.

[0246] In some embodiments, the second end is represented by formula (11-A):

[0247] [ka] or a pharmaceutically acceptable salt thereof.

[0248] In some embodiments, the second end is represented by formula (12-A):

[0249] [ka] or a pharmaceutically acceptable salt thereof, During the ceremony, A 4 Ha-CR 40 R 40 -or-NR 40 - and R 40 each independently represents hydrogen or an optionally substituted C-C 10 is alkyl, R 36 is an optionally substituted 5-6 membered heteroaryl; R 37 are each independently hydrogen, halogen, C1-C6 alkyl, or C1-C6 haloalkyl; R 38 is optionally substituted C1-C 10 alkyl, optionally substituted C3-C8 cycloalkyl, or optionally substituted 3- to 8-membered heterocycloalkyl; R 39are hydrogen, halogen, -OH, -CN, -NO2, -NH2, oxo (=O), =S, C1-C 10 Haloalkyl, or C1-C 10 is hydroxyalkyl, p 11 is 1 to 4, q1 and q2 each independently represent 0 to 2; The linker is R 38 or R 40 It is connected to formula (12-A) by either

[0250] In some embodiments, R 36 is an optionally substituted 5-membered heteroaryl. In some embodiments, R 36 is an optionally substituted oxazole, oxadiazole, thiazole, thiadiazole, pyrrole, or pyrazole. In some embodiments, R 36 is an optionally substituted oxazole.

[0251] In some embodiments, R 37 are each independently halogen, C-C alkyl, or C-C haloalkyl. 37 are each independently a halogen.

[0252] In some embodiments, R 38 is an optionally substituted C1-C 10 In some embodiments, R 38 is an optionally substituted C3-C8 cycloalkyl or an optionally substituted 3-8 membered heterocycloalkyl. In some embodiments, R 38 is a 3- to 8-membered heterocycloalkyl.

[0253] In some embodiments, R 39 are hydrogen, halogens, -OH, -CN, -NO2, -NH2, C1-C 10 Haloalkyl, or C1-C 10In some embodiments, R 39 is oxo or ═S. In some embodiments, R 39 is oxo. In some embodiments, R 39 is =S.

[0254] In some embodiments, A 4 Ha-NR 40 In some embodiments, A 4 is —NH. In some embodiments, A 6 is -NCH3. In some embodiments, A 4 Ha-CR 40 R 40 In some embodiments, A 4 is -CH2-.

[0255] In some embodiments, R 40 each independently represents an optionally substituted C-C 10 In some embodiments, R 40 are each independently hydrogen.

[0256] In some embodiments, p 11 is 3 or 4. In some embodiments, p 11 is 2. In some embodiments, p 11 is 1.

[0257] In some embodiments, q1 is 1 and q2 is 1. In some embodiments, q1 is 2 and q2 is 0.

[0258] In some embodiments, the linker is R 38 In some embodiments, the linker is linked to formula (12-A) via R 40 is connected to formula (12-A) via

[0259] In some embodiments, the second end is represented by formula (12-B) or formula (12-C):

[0260] [ka] or a pharmaceutically acceptable salt thereof.

[0261] In some embodiments, the second end is represented by formula (12-D) or formula (12-E):

[0262] [ka] or a pharmaceutically acceptable salt thereof.

[0263] In some embodiments, the second end is represented by formula (13-A):

[0264] [ka] or a pharmaceutically acceptable salt thereof, During the ceremony, Ring J is absent or is an optionally substituted 5-6 membered heteroaryl; R 41 is optionally substituted C1-C6 alkyl, optionally substituted C3-C8 cycloalkyl, -C(O)R 41a , -C(O)-, or -C(O)NR 41a R 41b and R 41a and R 41b each independently represents an optionally substituted C-C 10 alkyl or optionally substituted C3-C8 cycloalkyl; R 42 is an optionally substituted C1-C 10 Alkyl, optionally substituted C-C 10 haloalkyl, optionally substituted C3-C8 cycloalkyl, or optionally substituted 3- to 8-membered heterocycloalkyl; R 43 is hydrogen or optionally substituted C1-C 10 is alkyl, R 44 are each independently hydrogen, halogen, -OH, -CN, -NO2, -NH2, optionally substituted C1-C 10 Alkyl, optionally substituted C-C 10 Haloalkyl, C1-C 10 Hydroxyalkyl, optionally substituted C-C 10 Alkenyl, optionally substituted C-C 10 alkynyl, optionally substituted C3-C8-cycloalkyl, or optionally substituted 3- to 8-membered heterocycloalkyl; or R 43 and R 44 together with the atom to which they are attached form an optionally substituted 5- to 8-membered heterocycloalkyl; P 12 is 1 to 4, q3 is 0 or 1, Formula (13-A) is a ring J or R 41 is connected to the linker.

[0265] In some embodiments, R 41 is optionally substituted C1-C6 alkyl or optionally substituted C3-C8 cycloalkyl. In some embodiments, R 41 HA-C(O)R 41a In some embodiments, R 41 is —C(O)CH or —C(O)CHCH. In some embodiments, R 41 -C(O)-NR 41a R 41b is.

[0266] In some embodiments, R 41a is an optionally substituted C1-C 10 In some embodiments, R 41ais an optionally substituted C3-C8 cycloalkyl.

[0267] In some embodiments, R 41b is an optionally substituted C1-C 10 In some embodiments, R 41b is an optionally substituted C3-C8 cycloalkyl.

[0268] In some embodiments, R 42 is an optionally substituted C1-C 10 Alkyl or optionally substituted C-C 10 In some embodiments, R 42 is an optionally substituted C3-C8 cycloalkyl or an optionally substituted 3-8 membered heterocycloalkyl. In some embodiments, R 42 is an optionally substituted 3-8 membered heterocycloalkyl ring.

[0269] In some embodiments, R 43 is an optionally substituted C1-C 10 In some embodiments, R 43 is hydrogen.

[0270] In some embodiments, R 44 each independently represents a halogen, -OH, -CN, -NO2, -NH2, optionally substituted C1-C 10 Alkyl, optionally substituted C-C 10 Haloalkyl, C1-C 10 Hydroxyalkyl, optionally substituted C-C 10 Alkenyl, optionally substituted C-C 10 alkynyl, optionally substituted C3-C8-cycloalkyl, or an optionally substituted 3- to 8-membered heterocycle. 44 are each independently a halogen or C1-C 10 It is haloalkyl.

[0271] In some embodiments, R 43 and R 44 and one of R together with the atom to which they are attached form an optionally substituted 5-8 membered heterocycloalkyl. 43 and R 44 together with the atom to which they are attached form a 5-, 6-, 7-, or 8-membered heterocycloalkyl.

[0272] In some embodiments, p 12 is 3 or 4. In some embodiments, p 12 is 2. In some embodiments, p 12 is 1.

[0273] In some embodiments, q3 is 1. In some embodiments, q3 is 0.

[0274] In some embodiments, ring J is an optionally substituted 5-membered heteroaryl. In some embodiments, ring J is absent.

[0275] In some embodiments, Formula (13-A) is connected to a linker at ring J. In some embodiments, Formula (13-A) is connected to a linker at ring J. 41 is connected to the linker.

[0276] In some embodiments, the second end is represented by formula (13-B):

[0277] [ka] or a pharmaceutically acceptable salt thereof.

[0278] In some embodiments, the second end has the formula (13-C1) or (13-C2):

[0279] [ka] or a pharmaceutically acceptable salt thereof.

[0280] In some embodiments, the second end has formula (13-D1) or (13-D2):

[0281] [ka] or a pharmaceutically acceptable salt thereof.

[0282] In some embodiments, the second end is represented by formula (14-A):

[0283] [ka] or a pharmaceutically acceptable salt thereof, During the ceremony, Ring K is a 5- to 6-membered heterocycloalkyl; A 5 is absent, -CH2, -NH-, or -O-; L 4 is alkylene or heteroalkylene, R 45 each independently represents a halogen, -OH, -CN, -NO2, -NH2, optionally substituted C1-C 10 Alkyl, optionally substituted C-C 10 Haloalkyl, optionally substituted C-C 10 Hydroxyalkyl, optionally substituted C-C 10 Alkenyl, optionally substituted C-C 10 alkynyl, optionally substituted C3-C8-cycloalkyl, or optionally substituted 3- to 8-membered heterocycloalkyl, R 46 are each independently hydrogen, halogen, -OH, -CN, -NO2, -NH2, optionally substituted C1-C 10Alkyl, optionally substituted C-C 10 Haloalkyl, optionally substituted C-C 10 Heteroalkyl, optionally substituted C-C 10 Hydroxyalkyl, optionally substituted C-C 10 Alkenyl, optionally substituted C-C 10 alkynyl, optionally substituted C3-C8 cycloalkyl, or optionally substituted 3- to 8-membered heterocycloalkyl; R 47 is optionally substituted C1-C 10 Alkyl, -C(O)R 47a , or -C(O)-NR 47a R 47b and R 47a and R 47b each independently represents an optionally substituted C-C 10 alkyl or optionally substituted C3-C8 cycloalkyl; q4 is 2 to 3, or q5 is 0 to 2, Formula (14-A) can be formed via ring K or R 45 is connected to the linker via one of the

[0284] 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-.

[0285] 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, L4 is -O-CH2- or -O-CH2CH2-.

[0286] In some embodiments, R 45 each independently represents a halogen, -OH, -CN, -NO2, -NH2, optionally substituted C1-C 10 Alkyl, optionally substituted C-C 10 Haloalkyl, C1-C 10 Hydroxyalkyl, optionally substituted C-C 10 Alkenyl, or optionally substituted C-C 10 In some embodiments, R is alkynyl. 45 each independently represents an optionally substituted C-C 10 Alkyl or optionally substituted C-C 10 In some embodiments, R 45 are independently C1-C 10 In some embodiments, R 45 are each independently -OCH3 or -OCH2CH3.

[0287] In some embodiments, R 46 are each independently hydrogen, halogen, -OH, -CN, -NO2, -NH2, optionally substituted C1-C 10 Alkyl, optionally substituted C-C 10 Haloalkyl, optionally substituted C-C 10 Heteroalkyl, or optionally substituted C-C 10 In some embodiments, R 46 are each independently -OCH3. In some embodiments, R 46 are each independently hydrogen.

[0288] In some embodiments, R 47 is an optionally substituted C1-C 10 In some embodiments, R47 HA-C(O)R 47a In some embodiments, R 47 is —C(O)CH or —C(O)CHCH. In some embodiments, R 47 -C(O)-NR 47a R 47b is.

[0289] In some embodiments, R 47a is an optionally substituted C1-C 10 In some embodiments, R 47a is an optionally substituted C3-C8 cycloalkyl.

[0290] In some embodiments, R 47b is an optionally substituted C1-C 10 In some embodiments, R 47b is an optionally substituted C3-C8 cycloalkyl.

[0291] In some embodiments, ring K is a 6-membered heterocycloalkyl.

[0292] In some embodiments, q4 is 3. In some embodiments, q4 is 2.

[0293] In some embodiments, q5 is 2. In some embodiments, q5 is 1. In some embodiments, q5 is 0.

[0294] In some embodiments, Formula (14-A) is connected to the linker via ring K. In some embodiments, Formula (14-A) is R 45 is connected to the linker via one of the

[0295] In some embodiments, the second end is represented by formula (14-B) or formula (14-C):

[0296] [ka] or a pharmaceutically acceptable salt thereof.

[0297] In some embodiments, the second end is represented by formula (15-A):

[0298] [ka] or a pharmaceutically acceptable salt thereof, During the ceremony, Ring L is aryl or heteroaryl; R 48 are hydrogen, halogen, -OH, -CN, -NO2, -NH2, C1-C 10 Alkyl, C1-C 10 Haloalkyl, or C1-C 10 is hydroxyalkyl, R 49 and R 50 each independently represents hydrogen, optionally substituted C-C 10 Alkyl, optionally substituted C-C 10 Haloalkyl, optionally substituted C-C 10 Alkenyl, or optionally substituted C-C 10 is alkynyl, R 51 is hydrogen, halogen, -OH, -CN, -NO2, -NH2, optionally substituted C1-C 10 Alkyl, optionally substituted C-C 10 Haloalkyl, optionally substituted C-C 10 Hydroxyalkyl, optionally substituted C-C 10 Alkenyl, or optionally substituted C-C 10 is alkynyl, R 52 is hydrogen, halogen, -OH, -CN, -NO2, -NH2, optionally substituted C1-C 10 Alkyl, optionally substituted C-C 10Haloalkyl, or optionally substituted C-C 10 is hydroxyalkyl, p7 is 1 to 4.

[0299] In some embodiments, ring L is aryl. In some embodiments, aryl is phenyl. In some embodiments, ring L is heteroaryl. In some embodiments, ring L is a bicyclic heteroaryl containing 1, 2, or 3 heteroatoms selected from N and O.

[0300] In some embodiments, the second end is represented by formula (15-B):

[0301] [ka] or a pharmaceutically acceptable salt thereof, During the ceremony, R 48 are hydrogen, halogen, -OH, -CN, -NO2, -NH2, C1-C 10 Alkyl, C1-C 10 Haloalkyl, or C1-C 10 is hydroxyalkyl, R 49 and R 50 each independently represents hydrogen, optionally substituted C-C 10 Alkyl, optionally substituted C-C 10 Haloalkyl, optionally substituted C-C 10 Alkenyl, or optionally substituted C-C 10 is alkynyl, R 51 is hydrogen, halogen, -OH, -CN, -NO2, -NH2, optionally substituted C1-C 10 Alkyl, optionally substituted C-C 10 Haloalkyl, optionally substituted C-C 10 Hydroxyalkyl, optionally substituted C-C 10 Alkenyl, or optionally substituted C-C10 is alkynyl, p7 is 1 to 4.

[0302] In some embodiments, the second end is represented by formula (15-C):

[0303] [ka] or a pharmaceutically acceptable salt thereof, During the ceremony, X is CR 48 or N, R 48 are hydrogen, halogen, -OH, -CN, -NO2, -NH2, C1-C 10 Alkyl, C1-C 10 Haloalkyl, or C1-C 10 is hydroxyalkyl, R 49 and R 50 each independently represents hydrogen, optionally substituted C-C 10 Alkyl, optionally substituted C-C 10 Haloalkyl, optionally substituted C-C 10 Alkenyl, or optionally substituted C-C 10 is alkynyl, R 51 is hydrogen, halogen, -OH, -CN, -NO2, -NH2, optionally substituted C1-C 10 Alkyl, optionally substituted C-C 10 Haloalkyl, optionally substituted C-C 10 Hydroxyalkyl, optionally substituted C-C 10 Alkenyl, or optionally substituted C-C 10 is alkynyl, R 53 is hydrogen or optionally substituted C1-C 10 is alkyl, p7 is 1 to 3.

[0304] In some embodiments, R 48are hydrogen, halogen, -OH, -CN, -NO2, -NH2, C1-C 10 Alkyl, C1-C 10 Haloalkyl, or optionally substituted C-C 10 It is a hydroxyalkyl.

[0305] In some embodiments, R 49 is hydrogen, optionally substituted C-C 10 Alkyl, optionally substituted C-C 10 Haloalkyl, optionally substituted C-C 10 Alkenyl, or optionally substituted C-C 10 In some embodiments, R is alkynyl. 49 is optionally substituted C1-C 10 In some embodiments, R 49 is methyl, ethyl, iso-propyl, or tert-butyl. In some embodiments, R 49 is hydrogen.

[0306] In some embodiments, R 50 is hydrogen, optionally substituted C-C 10 Alkyl, optionally substituted C-C 10 Haloalkyl, optionally substituted C-C 10 Alkenyl, or optionally substituted C-C 10 In some embodiments, R is alkynyl. 50 is an optionally substituted C1-C 10 Alkyl or optionally substituted C-C 10 In some embodiments, R is alkenyl. 50 is hydrogen.

[0307] In some embodiments, R 51 is hydrogen, halogen, -OH, -CN, -NO2, -NH2, optionally substituted C1-C 10 Alkyl, optionally substituted C-C 10Haloalkyl, optionally substituted C-C 10 Hydroxyalkyl, optionally substituted C-C 10 Alkenyl, or optionally substituted C-C 10 It is alkynyl.

[0308] In some embodiments, R 52 is hydrogen, halogen, -OH, -CN, -NO2, -NH2, optionally substituted C1-C 10 Alkyl, optionally substituted C-C 10 Haloalkyl, or optionally substituted C-C 10 In some embodiments, R 52 is hydrogen.

[0309] In some embodiments, R 53 is hydrogen or optionally substituted C-C 10 In some embodiments, R 53 is optionally substituted C1-C 10 In some embodiments, R 53 is methyl, ethyl, iso-propyl, or tert-butyl. In some embodiments, R 53 is hydrogen.

[0310] In some embodiments, p7 is 4. In some embodiments, p7 is 3. In some embodiments, p7 is 2. In some embodiments, p7 is 1.

[0311] In some embodiments, the second end is a group represented by formula (15-D1), formula (15-D2), or formula (15-D3):

[0312] [ka] or a pharmaceutically acceptable salt thereof.

[0313] In some embodiments, the second end is a group represented by formula (15-E1), formula (15-E2), or formula (15-E3):

[0314] [ka] or a pharmaceutically acceptable salt thereof.

[0315] In some embodiments, the second end is represented by formula (16-A):

[0316] [ka] or a pharmaceutically acceptable salt thereof, During the ceremony, B 5 is -O-, -NH-, or S, B 6 is N or CH, R 54 is optionally substituted aryl or optionally substituted heteroaryl; R 55 each independently represents a halogen, -OH, -CN, -NO2, -NH2, optionally substituted C1-C 10 Alkyl, optionally substituted C-C 10 Haloalkyl, optionally substituted C-C 10 Heteroalkyl, or optionally substituted C-C 10 is hydroxyalkyl, R 56 is hydrogen, optionally substituted C1-C 10 Alkyl, optionally substituted C-C 10 Haloalkyl, or optionally substituted C-C 10 is hydroxyalkyl, R 57 is halogen, -OH, -CN, -NO2, -NH2, optionally substituted C1-C 10 Alkyl, optionally substituted C-C 10 Haloalkyl, or optionally substituted C-C10 is hydroxyalkyl, p9 is 1 to 3, q7 is 0 to 2.

[0317] In some embodiments, B 5 is —O— or —S—. In some embodiments, B 5 is —O—. In some embodiments, B 5 is -S-.

[0318] In some embodiments, B 6 is N. In some embodiments, B 6 is CH.

[0319] In some embodiments, R 54 is optionally substituted aryl. In some embodiments, R 54 is one or more halogens, -CN, -NH2, -OH, C1-C 10 Alkyl, C1-C 10 Haloalkyl, or C1-C 10 It is phenyl optionally substituted with hydroxyalkyl.

[0320] In some embodiments, R 55 each independently represents a halogen, —OH, —CN, —NH, optionally substituted C-C 10 Alkyl, optionally substituted C-C 10 Haloalkyl, optionally substituted C-C 10 Heteroalkyl, or optionally substituted C-C 10 It is a hydroxyalkyl.

[0321] In some embodiments, R 56 is an optionally substituted C1-C 10 Alkyl, optionally substituted C-C 10 Haloalkyl, or optionally substituted C-C 10 In some embodiments, R56 is optionally substituted C1-C 10 It is alkyl.

[0322] In some embodiments, R 57 is a halogen, -OH, -CN, -NO2, -NH2, or optionally substituted C1-C 10 It is alkyl.

[0323] In some embodiments, p9 is 3. In some embodiments, p9 is 2. In some embodiments, p9 is 1.

[0324] In some embodiments, q7 is 2. In some embodiments, q7 is 1. In some embodiments, q7 is 0.

[0325] In some embodiments, the second end is represented by formula (16-B):

[0326] [ka] or a pharmaceutically acceptable salt thereof.

[0327] In some embodiments, the second end is represented by formula (17-A):

[0328] [ka] or a pharmaceutically acceptable salt thereof, During the ceremony, Ring M is an optionally substituted aryl or an optionally substituted heteroaryl; Ring N is absent or is a 4- to 8-membered heterocycloalkyl; A 6 is -O-, -NH-, or -CH2-, R 58 are each independently halogen, -OH, -CN, -NO2, -NH2, C1-C 10Alkyl, C1-C 10 Haloalkyl, or C1-C 10 is a hydroxyalkyl, R 59 are hydrogen, -OH, -NH2, C1-C 10 Alkyl, C1-C 10 Haloalkyl, C1-C 10 Hydroxyalkyl, or -NH-C1-C 10 is alkyl, R 60 is hydrogen or optionally substituted C1-C 10 is alkyl, p 10 is 1 to 4 Equation (17-A) is R 59 is connected to the linker via

[0329] In some embodiments, ring M is one or more of halogen, CN, NH, OH, C-C 10 Alkyl, C1-C 10 Haloalkyl, or C1-C 10 In some embodiments, ring M is aryl optionally substituted with hydroxyalkyl. In some embodiments, ring M is phenyl. In some embodiments, ring M is one or more of halogen, CN, NH, OH, C-C 10 Alkyl, C1-C 10 Haloalkyl, or C1-C 10 and optionally substituted 6-membered heteroaryl optionally substituted with hydroxyalkyl. In some embodiments, ring M is an optionally substituted pyridine.

[0330] In some embodiments, Ring N is a 4-8 membered heterocycloalkyl. In some embodiments, Ring N is a 4 membered heterocycloalkyl. In some embodiments, Ring N is a 5 membered heteroaryl. In some embodiments, Ring N is a 6 membered heteroaryl. In some embodiments, Ring N is absent.

[0331] In some embodiments, A 6is —O— or —NH—. In some embodiments, A 6 is -CH2-.

[0332] In some embodiments, R 58 are each independently -OH, -NH2, C1-C 10 Alkyl, C1-C 10 Haloalkyl, or C1-C 10 In some embodiments, R 58 are independently C1-C 10 Alkyl or C1-C 10 In some embodiments, R 58 are independently C1-C 10 It is a hydroxyalkyl.

[0333] In some embodiments, R 59 -OH, -NH2, C1-C 10 Hydroxyalkyl, or -NH-C1-C 10 In some embodiments, R 59 is hydrogen.

[0334] In some embodiments, R 60 is an optionally substituted C1-C 10 In some embodiments, R 60 is methyl. In some embodiments, R 60 is hydrogen.

[0335] In some embodiments, p 10 is 3 or 4. In some embodiments, p 10 is 2. In some embodiments, p 10 is 1.

[0336] In some embodiments, the second end is represented by formula (17-B):

[0337] [ka] or a pharmaceutically acceptable salt thereof.

[0338] In some embodiments, the second end is represented by formula (17-C):

[0339] [ka] or a pharmaceutically acceptable salt thereof.

[0340] In some embodiments, the second end is

[0341] [ka]

[0342] [ka] or a pharmaceutically acceptable salt thereof.

[0343] Oligomeric Linker

[0344] The length of the oligomeric linker also depends on the type of regulatory protein and the type of 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 20-30 angstroms. In some embodiments, the oligomeric linker contains 5-50 chain atoms.

[0345] In some embodiments, the oligomeric linker comprises a multimer having between 2 and 50 spacer moieties, The spacer moieties are each independently -((CR 1b R 1b ) x -O) y -,-((CR 1b R 1b ) x -NR1a ) y -,-((CR 1b R 1b ) x -CH=CH-(CR 1b R 1b ) x -O) y -, optionally substituted C1-C 12 Alkyl, optionally substituted C-C 10 Alkenyl, optionally substituted C-C 10 Alkynyl, optionally substituted C-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 1a selected from the group consisting of -S(O)-, and -P(O)OH-, and any combination thereof; x's are independently 2 to 4; each y is independently 1 to 10; R 1a are each independently hydrogen or optionally substituted C1-C6 alkyl; R 1b are each 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 alkylamido, sulfonyl, optionally substituted thioalkoxy, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted cycloalkyl, and optionally substituted heterocyclyl.

[0346] In some embodiments, the oligomeric linker comprises a multimer having 2 to 50 spacer moieties, each independently an optionally substituted C-C 12 Alkyl, -((CH2) X -O) y -, -((CH2) x -NH) y -, -O-, -C(O)NH-, -NH-, and any combination thereof.

[0347] In various embodiments, a composition suitable for treating a genetic disease comprising a double-stranded DNA binding moiety 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 comprises an isotonicity adjusting agent, e.g., sodium chloride, a buffer, a stabilizer, an antioxidant, a viscosity enhancing agent, a solubilizing agent, a penetration enhancer, and / or a preservative. In some embodiments, the 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.

[0348] In various embodiments, administration of a composition suitable for treating genetic diseases that comprises a double-stranded DNA-binding moiety to the eye results in minimal systemic exposure. In some embodiments, local administration of the composition to the eye (e.g., ocular administration) results in the 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 penetrates the internal cells and tissues of the eye.

[0349] Aqueous dose vs. dose uniformity Typical ophthalmic aqueous solutions, emulsions, or suspensions are packaged in eye droppers and administered as drops. For example, a single administration (i.e., a single dose) of an ophthalmic aqueous solution, emulsion, or suspension comprises one, two, three, or more drops into a patient's eye. In some embodiments, a single 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 dropper.

[0350] In some cases, the compositions described herein include aqueous ophthalmic compositions that provide a uniform concentration per dose. In some instances, the uniform concentration per dose does not exhibit significant variations in drug content from one dose to another. In some instances, the uniform concentration per dose provides a consistent drug content from one dose to another.

[0351] In some embodiments, the composition has a dose-to-dose ophthalmic agent concentration variation of less than 50%. In some embodiments, the composition has a dose-to-dose ophthalmic agent concentration variation of less than 40%. In some embodiments, the composition has a dose-to-dose ophthalmic agent concentration variation of less than 30%. In some embodiments, the composition has a dose-to-dose ophthalmic agent concentration variation of less than 20%. In some embodiments, the composition has a dose-to-dose ophthalmic agent concentration variation of less than 10%. In some embodiments, the composition has a dose-to-dose ophthalmic agent concentration variation of less than 5%.

[0352] In some embodiments, the concentration variation of the ophthalmic agent per dose is based on 10 consecutive doses. In some embodiments, the concentration variation of the ophthalmic agent per dose is based on 8 consecutive doses. In some embodiments, the concentration variation of the ophthalmic agent per dose is based on 5 consecutive doses. In some embodiments, the concentration variation of the ophthalmic agent per dose is based on 3 consecutive doses. In some embodiments, the concentration variation of the ophthalmic agent per dose is based on 2 consecutive doses.

[0353] Non-settling compositions should not require shaking to uniformly distribute the drug. "No-shake" compositions are potentially advantageous over compositions requiring shaking for the simple reason that patient shaking behavior is a major source of variability in the amount of drug administered. It has been reported that patients often do not shake or forget to shake compositions requiring shaking before administering a dose, despite clear instructions to do so on the label. On the other hand, even patients who do shake the product generally cannot determine whether the strength and / or duration of shaking is adequate to achieve product uniformity. In some embodiments, the ophthalmic gel and ointment compositions described herein are "no-shake" formulations that maintain the dose-to-dose uniformity described herein. In some embodiments, the ophthalmic gel and ointment compositions described herein require shaking to maintain the dose-to-dose uniformity described herein.

[0354] To assess dose-to-dose uniformity, eye drop bottles or tubes containing the aqueous ophthalmic composition, ophthalmic gel composition, or ophthalmic ointment composition are stored upright for a minimum of 12 hours before the start of testing. To simulate the recommended dosing of these products, a predetermined number of drops or strips are dispensed from each commercial bottle or tube at predetermined time intervals over an extended period of time, or until no product remains in the commercial bottle or tube. All drops and strips are dispensed into tared glass bottles, 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.

[0355] aqueous viscosity In some embodiments, the composition is heated to about 20° C. and 1 s -1 In some embodiments, the composition has a Brookfield RVDV viscosity of about 1 to about 50,000 centipoise (cps) at a shear rate of about 20° C. and 1 s. -1In some embodiments, the composition has a Brookfield RVDV viscosity of about 100 to about 40,000 cps at a shear rate of about 20° C. and 1 s -1 In some embodiments, the composition has a Brookfield RVDV viscosity of about 500 to about 30,000 cps at a shear rate of about 20° C. and 1 s -1 In some embodiments, the composition has a Brookfield RVDV viscosity of about 1000 to about 20,000 cps at a shear rate of about 20° C. and 1 s. -1 In some embodiments, the composition has a Brookfield RVDV viscosity of about 2000 to about 10,000 cps at a shear rate of about 20° C. and 1 s. -1 and a Brookfield RVDV viscosity of about 4000 to about 8000 cps at a shear rate of about 1000 cps.

[0356] In some embodiments, the aqueous composition contains sufficient viscosity enhancer to provide a viscosity of about 500-50,000 cps, about 750-50,000 cps, about 1000-50,000 cps, about 1000-40,000 cps, about 2000-30,000 cps, about 3000-20,000 cps, about 4000-10,000 cps, or about 5000-8000 cps.

[0357] In some embodiments, the aqueous composition comprises a viscosity-lowering agent. In some embodiments, the viscosity-lowering agent comprises caffeine. In some embodiments, the aqueous composition comprises caffeine at a concentration appropriate to achieve a 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.

[0358] In some embodiments, the compositions described herein have a low viscosity at body temperature. In some embodiments, the low-viscosity composition comprises about 1% to about 10% of a viscosity-enhancing agent (e.g., a gelling component such as a polyoxyethylene-polyoxypropylene copolymer). In some embodiments, the low-viscosity composition comprises about 2% to about 10% of a viscosity-enhancing agent (e.g., a gelling component such as a polyoxyethylene-polyoxypropylene copolymer). In some embodiments, the low-viscosity composition comprises about 5% to about 10% of a viscosity-enhancing agent (e.g., a gelling component such as a polyoxyethylene-polyoxypropylene copolymer). In some embodiments, the low-viscosity composition is substantially free of a viscosity-enhancing agent (e.g., a gelling component such as a polyoxyethylene-polyoxypropylene copolymer). In some embodiments, the low-viscosity ophthalmic pharmaceutical composition described herein provides an apparent viscosity of about 100 cps to about 10,000 cps. In some embodiments, the low-viscosity ophthalmic pharmaceutical composition described herein provides an apparent viscosity of about 500 cps to about 10,000 cps. In some embodiments, the low viscosity ophthalmic pharmaceutical compositions described herein provide an apparent viscosity of about 1000 cps to about 10,000 cps.

[0359] osmolality 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.

[0360] As used herein, "practical osmolarity" or "deliverable osmolarity" refers to the osmolarity of a composition as determined by measuring the osmolarity of all excipients except the ophthalmic agent and gelling and / or thickening agents (e.g., polyoxyethylene-polyoxypropylene copolymer, carboxymethylcellulose, etc.). The practical osmolarity of a composition disclosed herein is measured by a suitable method, for example, the freezing point depression method as described in Viegas et al., Int. J. Pharm., 1998, 160, 157-162. In some instances, the practical osmolarity of a composition disclosed herein is measured by vapor pressure osmometry (e.g., vapor pressure depression method), which allows for the measurement of the osmolarity of a composition at elevated temperatures. In some instances, the vapor pressure depression method allows for the measurement of the osmolality of a composition that includes a gelling agent (e.g., a thermoreversible polymer) at high temperature, where the gelling agent is in the form of a gel.

[0361] In some embodiments, the osmolality at the target site of action (e.g., the eye) is about the same as the delivered osmolality of the compositions described herein. In some embodiments, the compositions described herein have a deliverable osmolality of about 150 mOsm / L to about 500 mOsm / L, about 250 mOsm / L to about 500 mOsm / L, about 250 mOsm / L to about 350 mOsm / L, about 280 mOsm / L to about 370 mOsm / L, or about 250 mOsm / L to about 320 mOsm / L.

[0362] A practical osmolality of the compositions disclosed herein is about 100 mOsm / kg to about 1000 mOsm / kg, about 200 mOsm / kg to about 800 mOsm / kg, about 250 mOsm / kg to about 500 mOsm / kg, about 250 mOsmkg to about 320 mOsmkg, about 250 mOsmkg to about 350 mOsmkg, or about 280 mOsm / kg to about 320 mOsm / kg. In some embodiments, the compositions described herein have a practical osmolality of about 100 mOsm / L to about 1000 mOsm / L, about 200 mOsm / L to about 800 mOsm / L, about 250 mOsm / L to about 500 mOsm / L, about 250 mOsm / L to about 350 mOsm / L, about 250 mOsm / L to about 320 mOsm / L, or about 280 mOsm / L to about 320 mOsm / L.

[0363] In some embodiments, suitable tonicity agents include, but are not limited to, pharmaceutically acceptable sugars, salts, or any combination or mixture thereof, such as, but not limited to, dextrose, glycerin, mannitol, sorbitol, sodium chloride, and other electrolytes. In some examples, the tonicity agent is selected from sodium chloride, sodium nitrate, sodium sulfate, sodium hydrogen sulfate, 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 a combination thereof.

[0364] In some embodiments, the compositions described herein contain one or more salts in an amount necessary to bring the osmolality of the composition into an acceptable range. Such salts include those having 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.

[0365] sterility In some embodiments, the compositions are sterilized. Means and processes for sterilization of the pharmaceutical compositions disclosed herein for human use are included in the embodiments disclosed herein. The goal is to provide safe pharmaceutical products that are relatively free of infectious disease-causing microorganisms. 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.

[0366] As used herein, sterilization refers to a process used to destroy or remove microorganisms present in a product or packaging. Any suitable method can be used to sterilize an object or composition. Methods available for inactivating microorganisms include, but are not limited to, the application of intense heat, lethal chemicals, or gamma radiation. In some embodiments, the process for 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 largely depends on the nature of the device or composition to be sterilized. A detailed description of many sterilization methods is provided in Chapter 40 of Remington: The Science and Practice of Pharmacy, published by Lippincott, Williams & Wilkins, which is incorporated by reference in this context.

[0367] filter sterilization Sterilizing filtration is a method used to remove microorganisms from a solution rather than destroy them. 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. Various filter membranes are used to selectively filter solutions with different properties. For example, PVF and PTFE membranes are well suited to filtering organic solvents, while aqueous solutions are filtered through PVF or MCE membranes. Filtration devices are available for use at many scales, from single-use, point-of-use disposable filters attached to syringes to commercial-scale filters used in manufacturing plants. Membrane filters are sterilized by autoclaving or chemical sterilization. The membrane filtration system was validated according to the following standardized protocol (Microbiological Evaluation of Filters for Sterilizing Liquids, Vol. 4, No. 3. Washington, DC: Health Industry Manufacturers Association, 1981). A known number of bacteria (approximately 10) such as Brevundimonas diminuta (ATCC 19146) was used. 7 / cm 2 ) into a membrane filter.

[0368] Pharmaceutical compositions are optionally sterilized by passing through a membrane filter. Compositions containing nanoparticles (U.S. Pat. No. 6,139,870) or multilamellar vesicles (Richard et al., International Journal of Pharmaceutics (2006), 312(1-2):144-50) are suitable for sterilization by filtration through a 0.22 μm filter without disrupting their organized structure.

[0369] In some embodiments, the methods disclosed herein include sterilizing the composition (or components thereof) by sterile filtration. For ophthalmic gel compositions comprising a thermosetting polymer, filtration is carried out at a temperature below the gel temperature (Tgel) of the compositions described herein (e.g., about 5°C) and at a viscosity that allows filtration within a reasonable time using a peristaltic pump (e.g., below the theoretical value of 100 cps).

[0370] Accordingly, provided herein are methods of sterilizing compositions provided herein that prevent degradation of polymeric components (e.g., thermosetting and / or other viscosity-enhancing agents) and / or ophthalmic agents during the sterilization process. In some embodiments, degradation of ophthalmic agents (e.g., double-stranded DNA-binding moieties) is reduced or eliminated by using a specific pH range for the buffer components in the composition and a specific ratio of viscosity-enhancing agent. In some embodiments, the selection of an appropriate viscosity-enhancing agent or thermosetting polymer allows for sterilization of the compositions described herein by filtration. In some embodiments, the use of an appropriate thermosetting polymer or other viscosity-enhancing agent in combination with a specific pH range of the composition allows for high-temperature sterilization of the described compositions without substantial degradation of the therapeutic agent or polymer excipients. An advantage of the sterilization methods provided herein is that, in certain instances, the compositions can be subjected to terminal sterilization via autoclaving and rendered substantially free of microorganisms and / or pyrogens without loss of the ophthalmic agent and / or excipients and / or viscosity-enhancing agents during the sterilization process.

[0371] Radiation sterilization One advantage of radiation sterilization is that it can sterilize many types of products without thermal degradation or other damage. A commonly used form of radiation is beta radiation, or alternatively, 60The gamma radiation is emitted from a CO source. Due to its penetrating ability, gamma radiation can be used to sterilize many types of products, including solutions, compositions, and heterogeneous mixtures. The germicidal effect of irradiation results from the interaction of gamma radiation with biological macromolecules. This interaction generates charged species and free radicals. Subsequent chemical reactions, such as rearrangement and cross-linking processes, cause the loss of normal function of these biological macromolecules. The compositions described herein are further optionally sterilized using beta radiation.

[0372] Heat sterilization Many methods are available for sterilization by the application of high heat. One method is the use of saturated steam autoclaves. In this method, saturated steam at a temperature of at least 121°C is contacted with the item to be sterilized. When an item is sterilized, heat is transferred directly to the microorganisms or indirectly to the microorganisms by heating the bulk aqueous solution to be sterilized. This method is widely practiced because it is a flexible, safe, and economical sterilization process.

[0373] microorganisms In some embodiments, the composition is substantially free of microorganisms. Acceptable bioburden or sterility levels include, but are not limited to, those specified by the United States Pharmacopeia. <1111> The acceptable sterility (e.g., bioburden) level is based on applicable standards for defining a therapeutically acceptable composition, including those listed in the following sections: 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 sterility of a composition include microbial agents with less than 10 cfu / mL, less than 50 cfu / mL, less than 500 cfu / mL, or less than 1000 cfu / mL. Additionally, acceptable bioburden levels or sterility include the exclusion of specified undesirable microbial agents. By way of example, specified undesirable microbial agents include, but are not limited to, Escherichia coli (E. coli), Salmonella spp., Pseudomonas aeruginosa (P. aeruginosa), and / or other specific microbial agents.

[0374] Sterility assurance: A key component of the quality control, quality assurance, and validation process is the method of sterility testing. Sterility testing is performed by two methods, by way of example only. The first method is direct inoculation, in which a sample of the composition being tested is added to growth medium and incubated for a period of up to 21 days. Turbidity in the growth medium indicates contamination. Disadvantages of this method include the small sampling size of the bulk material, which reduces sensitivity, and detection of microbial growth based on visual observation. Another method is membrane filtration sterility testing. In this method, a large volume of product is passed through a small membrane filter paper. The filter paper is then placed in a medium to promote microbial growth. This method has the advantage of being highly sensitive, since the entire bulk product is sampled. The commercially available Millipore Steritest sterility testing system is optionally used for membrane filtration sterility testing. For filtration testing of creams or ointments, the Steritest filtration system No. TLHVSL210 is used. For filtration testing of emulsions or sticky products, use Steritest filtration system No. TLAREM210 or TDAREM210. For filtration testing of pre-filled syringes, use Steritest filtration system No. TTHASY210. For filtration testing of substances dispensed as aerosols or foams, use Steritest filtration system No. TTHVA210. For filtration testing of dissolvable powders in ampoules or vials, use Steritest filtration system No. TTHADA210 or TTHADV210.

[0375] Tests for E. coli and Salmonella include the use of lactose broth incubated at 30-35°C for 24-72 hours, MacConkey and / or EMB agar for 18-24 hours, and / or Rappaport medium. Tests for the detection of Pseudomonas aeruginosa include the use of NAC agar. <62> The chapter further lists test procedures for specified objectionable microorganisms.

[0376] 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 a microbial agent per gram of formulation. In certain embodiments, the compositions described herein are formulated to be isotonic with the eye.

[0377] Endotoxin A further aspect of the sterilization process is the removal of by-products from the killing of microorganisms. The process of depyrogenation removes pyrogens from a sample. Pyrogens are endotoxins or exotoxins that trigger an immune response. One example of an endotoxin is the lipopolysaccharide (LPS) molecule found in the cell walls of gram-negative bacteria. Sterilization processes such as autoclaving or treatment with ethylene oxide kill bacteria, but LPS residues trigger pro-inflammatory immune responses such as septic shock. Because endotoxin molecular sizes vary widely, the presence of endotoxin is expressed in "endotoxin units" (EU). 1 EU is equivalent to 100 picograms of E. coli LPS. In some cases, humans may develop a response to as little as 5 EU / kg of body weight. Bioburden (e.g., microbial limit) and / or sterility (e.g., endotoxin level) may be expressed in any units as recognized in the art. In certain embodiments, the compositions described herein comprise lower endotoxin levels (e.g., less than 4 EU / kg of a subject's body weight) compared to conventionally acceptable endotoxin levels (e.g., 5 EU / kg of a subject's body weight). In some embodiments, the compositions have less than about 5 EU / kg of a subject's body weight. In other embodiments, the compositions have less than about 4 EU / kg of a subject's body weight. In further embodiments, the compositions have less than about 3 EU / kg of a subject's body weight. In further embodiments, the compositions have less than about 2 EU / kg of a subject's body weight.

[0378] In some embodiments, the compositions have less than about 5 EU / kg of composition. In other embodiments, the compositions have less than about 4 EU / kg of composition. In further embodiments, the compositions have less than about 3 EU / kg of composition. In some embodiments, the compositions have less than about 2 EU / kg of composition. In other embodiments, the compositions have less than about 1 EU / kg of composition. In further embodiments, the compositions have less than about 0.2 EU / kg of composition. In specific embodiments, the compositions described herein contain about 1 to about 5 EU / mL of composition. In specific embodiments, the compositions described herein contain about 2 to about 5 EU / mL of composition, about 3 to about 5 EU / mL of composition, or about 4 to about 5 EU / mL of composition.

[0379] In certain embodiments, the compositions described herein contain lower endotoxin levels (e.g., compositions of less than 0.5 EU / mL) compared to conventionally acceptable endotoxin levels (e.g., compositions of 0.5 EU / mL). In some embodiments, the compositions have a composition of less than about 0.5 EU / mL. In other embodiments, the compositions have a composition of less than about 0.4 EU / mL. In further embodiments, the compositions have a composition of less than about 0.2 EU / mL.

[0380] Pyrogen detection is accomplished by several methods, by way of example only. Suitable sterility tests are described in the United States Pharmacopeia (USP) <71> Includes tests described in Sterility Testing Methods (23rd edition, 1995). Both the rabbit pyrogen test and the Limulus amebocyte lysate test are described in the United States Pharmacopoeia. <85> Chapter and <151> Chapter (USP23 / NF 18, Biological Tests, The United States Pharmaceutical Convention, Rockville, MD, 1995). Alternative pyrogen assays have been developed based on monocyte-activating cytokine assays. Homogeneous cell lines suitable for quality control applications have been developed and have 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 a further embodiment, the composition is subjected to depyrogenation. In a further embodiment, the process of manufacturing the composition includes testing the composition for pyrogenicity. In certain embodiments, the compositions described herein are substantially free of pyrogens.

[0381] Ophthalmic gel composition Gels are defined in various ways. For example, the United States Pharmacopoeia defines a gel as a semi-solid system consisting of a suspension of either small inorganic particles or large organic molecules permeated with a liquid. Gels include single-phase or two-phase systems. Single-phase gels consist of organic macromolecules uniformly distributed throughout a liquid, with no clear boundary between the dispersed macromolecules and the liquid. Some single-phase gels are prepared from synthetic macromolecules (e.g., carbomer) or natural gums (e.g., tragacanth). In some embodiments, single-phase gels are typically aqueous, but can also be made using alcohols and oils. Two-phase gels consist of a network of small, discrete particles.

[0382] In some embodiments, gels are also classified as hydrophobic or hydrophilic gels. In certain embodiments, non-limiting examples of hydrophobic gel bases include liquid paraffin with polyethylene, or fatty oils gelled with colloidal silica, or aluminum or zinc soaps. In contrast, non-limiting examples of hydrophilic gel bases include water, glycerol, or propylene glycol gelled with a suitable gelling agent (e.g., tragacanth, starch, cellulose derivatives, carboxyvinyl polymers, and magnesium aluminum silicate). In certain embodiments, the rheology of the compositions disclosed herein is pseudoplastic, plastic, thixotropic, or expansive.

[0383] In some embodiments, the composition is an ophthalmic gel, and the ophthalmically acceptable carrier comprises water and at least one viscosity-enhancing agent. In some embodiments, the viscosity-enhancing agent is selected from a cellulose-based polymer, a polyoxyethylene-polyoxypropylene triblock copolymer, a dextran-based polymer, polyvinyl alcohol, dextrin, polyvinylpyrrolidone, a polyalkylene glycol, chitosan, collagen, gelatin, hyaluronic acid, or a combination thereof.

[0384] In some embodiments, the composition comprises a viscosity-lowering agent. In some embodiments, the viscosity-lowering agent comprises caffeine. In some embodiments, the composition comprises caffeine at a concentration appropriate to achieve a 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.

[0385] In some embodiments, the ophthalmic gel compositions described herein are in a semi-solid or gelled state (e.g., at room temperature) before topical administration. For example, suitable viscosity enhancers for such gels include, by way of example only, gelling agents and suspending agents. In one embodiment, the viscosity-enhanced composition does not contain a buffer. In another embodiment, the viscosity-enhanced composition contains a pharmaceutically acceptable buffer. If necessary, sodium chloride or other tonicity agents are optionally used to adjust tonicity.

[0386] By way of example only, ophthalmically acceptable viscosity agents include hydroxypropyl methylcellulose, hydroxyethyl cellulose, polyvinylpyrrolidone, carboxymethyl cellulose, polyvinyl alcohol, sodium chondroitin sulfate, or sodium hyaluronate. Other viscosity enhancing agents that are compatible with the targeted ocular site include, but are not limited to, acacia (gum arabic), agar, magnesium aluminum silicate, sodium alginate, sodium stearate, bladderwrack, bentonite, carbomer, carrageenan, Carbopol, xanthan, cellulose, microcrystalline cellulose (MCC), ceratonia, chitin, carboxymethyl chitosan, chondrus, dextrose, furcellaran, gelatin, ghatti gum, guar ... gum), guar gum, hectorite, lactose, sucrose, maltodextrin, mannitol, sorbitol, honey, corn starch, wheat starch, rice starch, potato starch, gelatin, gum Araya, xanthan gum, tragacanth gum, ethyl cellulose, ethyl hydroxyethyl cellulose, ethyl methyl cellulose, methyl cellulose, hydroxyethyl cellulose, hydroxyethyl methyl cellulose, hydroxypropyl cellulose, poly(hydroxyethyl methacrylate), oxypolygelatin, pectin, Examples of viscosity enhancing excipients include polygeline, povidone, propylene carbonate, methyl vinyl ether / maleic anhydride copolymer (PVM / MA), poly(methoxyethyl methacrylate), poly(methoxyethoxyethyl methacrylate), hydroxypropyl cellulose, hydroxypropylmethylcellulose (HPMC), sodium carboxymethylcellulose (CMC), silicon dioxide, polyvinylpyrrolidone (PVP: povidone), Splenda® (dextrose, maltodextrin, and sucralose), or combinations thereof.In certain embodiments, the viscosity enhancing excipient is a combination of MCC and CMC.In another embodiment, the viscosity enhancing excipient is a combination of carboxymethylated chitosan, or chitin, and alginate.The combination of chitin and alginate with the ophthalmic agent disclosed herein acts as a controlled release composition, limiting the diffusion of the ophthalmic agent from the composition. Additionally, the combination of carboxymethylated chitosan and alginate is optionally used to facilitate increased permeability of the ophthalmic agent in the eye.

[0387] In some embodiments, provided herein are viscosity-enhancing compositions comprising about 0.1 mM to about 100 mM of an ophthalmic agent, a pharmaceutically acceptable viscosity agent, and water for injection, wherein the concentration of the viscosity agent in the water is sufficient to provide the viscosity-enhancing composition with a final viscosity of about 100 to about 100,000 cps. In certain embodiments, the viscosity of the gel ranges from about 100 to about 50,000 cps, about 100 cps to about 1,000 cps, about 500 cps to about 1,500 cps, about 1,000 cps to about 3,000 cps, about 2,000 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 highly concentrated samples, the biocompatible viscosity-enhancing 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.

[0388] In one embodiment, the pharmaceutically acceptable viscosity-enhanced ophthalmically acceptable composition comprises at least one ophthalmic agent and at least one gelling agent. Suitable gelling agents for use in preparing the gel composition include, but are not limited to, cellulose, cellulose derivatives, cellulose ethers (e.g., carboxymethylcellulose, ethylcellulose, hydroxyethylcellulose, hydroxymethylcellulose, hydroxypropylmethylcellulose, hydroxypropylcellulose, methylcellulose), guar gum, xanthan gum, locust bean gum, alginates (e.g., alginic acid), silicates, starch, tragacanth, carboxyvinyl polymers, carrageenan, paraffin, petrolatum, and any combination or mixture thereof. In some other embodiments, hydroxypropylmethylcellulose (Methocel®) is used as a gelling agent. In certain embodiments, the clay enhancing agents described herein are also used as gelling agents for the gel compositions presented herein.

[0389] In some embodiments, the ophthalmic gel compositions described herein are in situ gel compositions. In some instances, the in situ gel compositions are based on increased precorneal residence time of the composition, which improves visual bioavailability, corneal mucoadhesion, lysosomal interactions and ionic gelation, improved corneal absorption, thermal gelation, or a combination thereof. In some instances, the in situ gel formulations are activated by pH, temperature, ions, UV, or solvent exchange.

[0390] In some examples, the ophthalmic gel composition comprises a double-stranded DNA binding moiety and one or more gelling agents. In some examples, the gelling agents include, but are not limited to, poloxamer (e.g., poloxamer 407), tetronics, ethyl (hydroxyethyl) cellulose, cellulose acetate phthalate (CAP), carbopol (e.g., Carbopol 1342P NF, Carbopol 980 NF), alginate (e.g., low-acetyl gellan gum (Gelrite®)), gellan, hyaluronic acid, Pluronic (e.g., Pluronic F-127), chitosan, polyvinyl alcohol (PVA), polyvinylpyrrolidone (PVP), dextran, hydroxypropylmethylcellulose (HPMC), hydroxyethylcellulose (HEC), methylcellulose (MC), thiolated xyloglucan, polymethacrylic acid (PMMA), polyethylene glycol (PEG), pseudolatex, xyloglucan, or a combination thereof.

[0391] In some examples, the in situ gel composition further comprises a penetration enhancer, which in some examples includes a surfactant (e.g., a non-ionic surfactant), benzalkonium chloride, EDTA, a surfactant heteroglycoside, a calcium chelator, hydroxylpropyl beta cyclodextrin (HP beta CD), a bile salt, or the like.

[0392] In some embodiments, other gel compositions may be useful depending on the particular ophthalmic agent, other pharmaceutical agent, or excipient / additive used, and are therefore considered to be within the scope of the present disclosure. For example, gelatin and its derivatives, alginate and alginate-based gels, and various natural and synthetic hydrogels and hydrogel-derived compounds, in addition to other commercially available glycerin-based gels, glycerin-derived compounds, conjugated or crosslinked gels, matrices, hydrogels, and polymers, are all expected to be useful in the ophthalmic agent compositions described herein. In some embodiments, ophthalmically acceptable gels include, but are not limited to, alginate hydrogel SAFR-gel (ConvaTec, Princeton, NJ), Duoderm® Hydroactive Gel (ConvaTec), Nu-gel® Johnson & Johnson Medical, Arlington, Tex.; Carrasyn® (V) acemannan hydrogel (Carrington Laboratories, Inc., Irving, Tex.); glycerin gel Elta® Hydrogel (Swiss-American Products, Inc., Dallas, Tex.), and KY® Sterile (Johnson & Johnson). In further embodiments, biodegradable, biocompatible gels also refer to compounds present in the ophthalmically acceptable compositions described and disclosed herein.

[0393] In some embodiments, the viscosity enhancing agent is a cellulosic polymer selected from cellulose gum, alkyl cellulose, hydroxyl-alkyl cellulose, hydroxyl-alkyl alkyl cellulose, carboxy-alkyl cellulose, or a combination thereof. In some embodiments, the viscosity enhancing agent is a hydroxyl-alkyl alkyl cellulose. In some embodiments, the viscosity enhancing agent is hydroxypropyl methylcellulose.

[0394] In certain embodiments, the viscosity-enhancing composition is characterized by a phase transition between room temperature and body temperature (e.g., including in individuals with a severe fever up to about 42°C). In some embodiments, the phase transition occurs at 1°C below body temperature, 2°C below body temperature, 3°C below body temperature, 4°C below body temperature, 6°C below body temperature, 8°C below body temperature, or 10°C below body temperature. In some embodiments, the phase transition occurs at about 15°C below body temperature, about 20°C below body temperature, or about 25°C below body temperature. In certain embodiments, the gelation temperature (Tgel) of the compositions described herein is about 20°C, about 25°C, or about 30°C. In certain embodiments, the gelation temperature (Tgel) of the compositions described herein is about 35°C or about 40°C. Included within the definition of body temperature is the body temperature of a healthy individual or an unhealthy individual, including an individual with a fever (up to 42°C). In some embodiments, the pharmaceutical compositions described herein are liquid at about room temperature and are administered at or about room temperature.

[0395] Copolymers of polyoxypropylene and polyoxyethylene (e.g., polyoxyethylene-polyoxypropylene triblock copolymers) form thermosetting gels when introduced into aqueous solutions. These polymers have the ability to change from a liquid to a gel state at temperatures close to body temperature, thus enabling useful compositions to be applied to targeted ocular sites. The phase transition from the liquid to the gel state depends on the polymer concentration and components in the solution.

[0396] In some embodiments, the amount of thermosetting polymer in any composition described herein is about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, or about 40% of the total weight of the composition. In some embodiments, the amount of thermosetting polymer in any composition 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 weight of the composition. In some embodiments, the amount of thermosetting polymer (e.g., Poloxamer 407) in any composition described herein is about 7.5% of the total weight of the composition. In some embodiments, the amount of thermosetting polymer (e.g., Poloxamer 407) in any composition described herein is about 10% of the total weight of the composition. In some embodiments, the amount of thermosetting polymer (e.g., poloxamer 407) in any composition described herein is about 11% by weight of the total composition. In some embodiments, the amount of thermosetting polymer (e.g., poloxamer 407) in any composition described herein is about 12% by weight of the total composition. In some embodiments, the amount of thermosetting polymer (e.g., poloxamer 407) in any composition described herein is about 13% by weight of the total composition. In some embodiments, the amount of thermosetting polymer (e.g., poloxamer 407) in any composition described herein is about 14% by weight of the total composition. In some embodiments, the amount of thermosetting polymer (e.g., poloxamer 407) in any composition described herein is about 15% by weight of the total composition. In some embodiments, the amount of thermosetting polymer (e.g., poloxamer 407) in any composition described herein is about 16% by weight of the total composition. In some embodiments, the amount of thermosetting polymer (e.g., poloxamer 407) in any composition described herein is about 17% by weight of the total composition. In some embodiments, the amount of thermosetting polymer (e.g., poloxamer 407) in any composition described herein is about 18% by weight of the total composition.In some embodiments, the amount of thermosetting polymer (e.g., poloxamer 407) in any composition described herein is about 19% by weight of the total composition. In some embodiments, the amount of thermosetting polymer (e.g., poloxamer 407) in any composition described herein is about 20% by weight of the total composition. In some embodiments, the amount of thermosetting polymer (e.g., poloxamer 407) in any composition described herein is about 21% by weight of the total composition. In some embodiments, the amount of thermosetting polymer (e.g., poloxamer 407) in any composition described herein is about 22% by weight of the total composition. In some embodiments, the amount of thermosetting polymer (e.g., poloxamer 407) in any composition described herein is about 23% by weight of the total composition. In some embodiments, the amount of thermosetting polymer (e.g., poloxamer 407) in any composition described herein is about 24% by weight of the total composition. In some embodiments, the amount of thermosetting polymer (e.g., poloxamer 407) in any composition described herein is about 25% by weight of the total composition. In some embodiments, the amount of thickening agent (e.g., gelling agent) in any composition described herein is about 1%, about 5%, about 10%, or about 15% by weight of the total composition. In some embodiments, the amount of thickening agent (e.g., gelling agent) in any composition 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% by weight of the total composition.

[0397] In an alternative embodiment, 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 sol-gel behavior over concentrations of about 5% w / w to about 40% w / w. Depending on the desired properties, the lactide / glycolide molar ratio 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 RESOMER RGP t50106, manufactured by Boehringer Ingelheim. This material is composed of a PLGA copolymer of 50:50 poly(DL-lactide-co-glycolide) with 10% w / w of PEG, and has a molecular weight of approximately 6000.

[0398] Additional biodegradable thermoplastic polyesters include AtriGel® (provided by Atrix Laboratories, Inc.) and / or those disclosed, for example, in U.S. Patent Nos. 5,324,519, 4,938,763, 5,702,716, 5,744,153, and 5,990,194, where suitable biodegradable thermoplastic polyesters are disclosed as thermoplastic polymers. Examples of suitable biodegradable thermoplastic polyesters include polylactides, polyglycosides, polycaprolactones, copolymers thereof, terpolymers thereof, and any combination thereof. In some such embodiments, the suitable biodegradable thermoplastic polyester is polylactide, polyglycosides, copolymers thereof, terpolymers thereof, or combinations thereof. In one embodiment, the biodegradable thermoplastic polyester is a 50 / 50 poly(DL-lactide-co-glycolide) with carboxy end groups, present in about 30% to about 40% by weight of the composition, and has an average molecular weight of about 23,000 to about 45,000. Alternatively, in another embodiment, the biodegradable thermoplastic polyester is a 75 / 25 poly(DL-lactide-co-glycolide) without carboxy end groups, present in about 40% to about 50% by weight of the composition, and has an average molecular weight of about 15,000 to about 24,000. In further or alternative embodiments, the end groups of the poly(DL-lactide-co-glycolide) are either hydroxyl, carboxyl, or ester, depending on the method of polymerization. Polycondensation of lactic acid or glycolic acid provides a polymer with terminal hydroxyl and carboxyl groups. Ring-opening polymerization of cyclic lactide or glycolide monomers with water, lactic acid, or glycolic acid results in polymers with the same end groups. However, ring-opening of cyclic monomers with monofunctional alcohols such as methanol, ethanol, or 1-dodecanol results in polymers with one hydroxyl group and one ester end group.Ring-opening polymerization of cyclic monomers with diols such as 1,6-hexanediol or polyethylene glycol results in polymers with only hydroxyl end groups.

[0399] Because the polymer system of a thermosetting gel dissolves more completely at the reduction temperature, the solubilization process involves adding the required amount of polymer to the amount of water used at the reduction temperature. Generally, the polymer is wetted by shaking, and then the mixture is capped and placed in a cooling chamber or constant-temperature container at approximately 0–10°C to dissolve the polymer. The mixture is stirred or shaken to induce more rapid dissolution of the thermosetting gel polymer. Ophthalmic medications and various additives, such as buffers, salts, and preservatives, are then added and dissolved. In some cases, pharmaceutical agents are suspended if they are insoluble in water. The pH is adjusted by adding an appropriate buffer.

[0400] Ophthalmic ointment composition Ointments are semisolid preparations of homogeneous viscosity intended for external application to the skin or mucous membranes, most commonly thick, greasy oils (e.g., 80% oil - 20% water). Ointments have a water number that defines the maximum amount of water they contain. Ointments are used as emollients or for the application of active ingredients to the skin for protective, therapeutic, or preventative purposes, and when a degree of occlusion is desired. Ointments are used topically on various body surfaces, including the skin and mucous membranes of the eye (eye ointment), vulva, anus, and nose.

[0401] The vehicle of an ointment is known as an ointment base. The choice of base depends on the clinical indication of the ointment. Various types of ointment bases include hydrocarbon bases such as hard paraffin, soft paraffin, microcrystalline wax, and ceresin, water-absorbing bases such as wool fat and beeswax, water-soluble bases such as macrogol 200, 300, and 400, emulsifying bases such as emulsifying wax, cetrimide, vegetable oils such as olive oil, coconut oil, sesame oil, almond oil, and peanut oil.

[0402] 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 further derived from hydrocarbon (fatty), absorbent, water-removable, or water-soluble bases. The active agent is dispersed in the base and then split after the drug penetrates the target site (e.g., membrane, skin, etc.).

[0403] The present disclosure recognizes that it is often difficult to incorporate low concentrations of drugs into ointments with sufficient dose-to-dose uniformity to effectively treat a disorder or disease. In some embodiments, poly(ethylene glycol), polyethoxylated castor oil (Cremophor® EL), alcohols having 12 to 20 carbon atoms, or mixtures of two or more of the foregoing, are effective excipients for dispersing and / or dissolving effective amounts of ophthalmic drugs. The resulting ointments are well tolerated by skin and ocular tissues.

[0404] The present disclosure further recognizes that ophthalmic drugs, such as double-stranded DNA-binding moieties incorporated into the ointment compositions described herein, are targeted to the choroid and / or retina of a patient when the composition is topically administered to the ocular surface, particularly the sclera of the patient. In some embodiments, the ophthalmic ointment composition comprises an ophthalmic drug, an ointment base, and an agent for dispersing and / or dissolving the drug in the ointment base, the agent being selected from poly(ethylene glycol), polyethoxylated castor oil, alcohols having 12 to 20 carbon atoms, and mixtures of two or more of the foregoing ingredients.

[0405] In some embodiments, the ointment base comprises an ophthalmically acceptable oil or fatty base, such as a natural wax, e.g., white and yellow beeswax, carnauba wax, wool wax (sheep's wool wax), purified lanolin, anhydrous lanolin, petroleum waxes, e.g., hard paraffin, microcrystalline wax, hydrocarbons, e.g., liquid paraffin, white and yellow soft paraffin, white petrolatum, yellow petrolatum, or combinations thereof.

[0406] The above mentioned oil bases are described in more detail, for example, in the British Pharmacopoeia 2001 Edition or the European Pharmacopoeia 3rd Edition.

[0407] In some embodiments, the ointment base is present in an amount of about 50 to about 95% by weight, preferably 70 to 90% by weight, based on the total weight of the composition.

[0408] Preferred ointment bases include one or more natural waxes such as those set forth above, preferably one or more of wool wax (wool wax), in combination with one or more hydrocarbons such as those set forth above, preferably soft paraffin or petrolatum, more preferably liquid paraffin.

[0409] A particular embodiment of the aforementioned ointment base comprises, for example, 5 to 17 parts by weight of wool fat, and 50 to 65 parts by weight of white petrolatum, and also 20 to 30 parts by weight of liquid paraffin.

[0410] In some embodiments, the agent for dispersing and / or dissolving the eye drop 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 the above ingredients. The agent is preferably used in an amount of 1 to 20% by weight, more preferably 1 to 10% by weight, of the total semi-solid composition.

[0411] Alcohols with 12 to 20 carbon atoms are especially stearyl alcohol (C 18 H 37 OH), cetyl alcohol (C 16 H 33OH), and mixtures thereof. Preferred are mixtures of solid alcohols consisting essentially of so-called cetostearyl alcohol, stearyl and cetyl alcohol, preferably containing 40% by weight or more of stearyl alcohol and up to at least 90% by weight of the sum of stearyl alcohol and cetyl alcohol, as well as compositions containing 80% by weight or more of cetostearyl alcohol and an emulsifier, in particular sodium cetostearyl sulfate and / or sodium lauryl sulfate, preferably in an amount of 7% by weight or more of emulsifier.

[0412] Polyethoxylated castor oil is a reaction product of natural or hardened castor oil and ethylene glycol. In some examples, such products can be obtained by reacting natural or hardened castor oil or a fraction thereof with ethylene oxide in a molar ratio of, for example, about 1:30 to about 1:60, with optional removal of free polyethylene glycol components from the product, according to known methods, for example, the methods disclosed in German Auslegeschriften 1,182,388 and 1,518,819. In particular, the product commercially available under the trade name Cremophor® EL, which has a molecular weight (measured by vapor osmometry) of about 1630, a saponification number of about 65-70, an acid number of about 2, an iodine number of about 28-32, and an nD25 of about 1.471, is suitable and preferred. For example, Nikkol® HCO-60, a reaction product of hydrogenated castor oil and ethylene oxide, exhibiting the following properties: acid number = about 0.3, saponification number = about 47.4, hydroxy number = about 42.5, pH (5%) = about 4.6, Color APHA = about 40, mp = about 36.0°C, freezing point = about 32.4°C, HO content (%, KF) = about 0.03, is suitable for use in this category.

[0413] Poly(ethylene glycol) is used in some embodiments as an agent for dispersing and / or dispersing eye drops in an ointment base in accordance with the present disclosure. Suitable poly(ethylene glycol)s typically have the general formula H—(OCH—CH) nThe poly(ethylene glycol) is a mixture of polymeric compounds of OH, where the index n is typically in the range of 4 to 230, and the average molecular weight is typically in the range of 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 1,000. In some embodiments, n is a number from about 6 to about 13, and the average molecular weight is about 300 to about 600. In some embodiments, n has a value from about 8.5 to about 9, and the relative molecular weight is about 400. Suitable poly(ethylene glycol)s, for example, poly(ethylene glycol)s having average molecular weights of about 200, 300, 400, 600, 1,000, 1,500, 2,000, 3,000, 4,000, 6,000, 8,000, and 10,000, are readily commercially available.

[0414] In some embodiments, poly(ethylene glycol), particularly of the type described in the preceding paragraph, is used in an amount of 1-10% or 1-5% by weight of the total semi-solid composition.

[0415] In some embodiments, the composition comprises an agent for dispersing and / or dissolving the drug in an ointment base selected from poly(ethylene-glycol), polyethoxylated castor oil, and preferably a mixture of the above ingredients.

[0416] Gel / ointment viscosity In some embodiments, the composition is heated to about 20° C. and 1 s -1 In some embodiments, the composition has a Brookfield RVDV viscosity of about 10,000 to about 300,000 cps at a shear rate of about 20° C. and 1 s. -1 In some embodiments, the composition has a Brookfield RVDV viscosity of about 15,000 to about 200,000 cps at a shear rate of about 20° C. and 1 s. -1 In some embodiments, the composition has a Brookfield RVDV viscosity of about 50,000 to about 150,000 cps at a shear rate of about 20° C. and 1 s. -1In some embodiments, the composition has a Brookfield RVDV viscosity of about 70,000 to about 130,000 cps at a shear rate of about 20° C. and 1 s. -1 and a Brookfield RVDV viscosity of about 90,000 to about 110,000 cps at a shear rate of about 100,000 cps.

[0417] In some embodiments, the ophthalmic gel composition contains sufficient viscosity enhancing agent to provide a viscosity of about 500-1,000,000 centipoise, about 750-1,000,000 centipoise, about 1,000-1,000,000 centipoise, about 1,000-400,000 centipoise, about 2,000-100,000 centipoise, about 3,000-50,000 centipoise, about 4,000-25,000 centipoise, about 5,000-20,000 centipoise, or about 6,000-15,000 centipoise. In some embodiments, the ophthalmic gel composition contains sufficient viscosity enhancing agent to provide a viscosity of about 500,000-1,000,000 centipoise.

[0418] In some embodiments, the compositions described herein are low-viscosity compositions at body temperature. In some embodiments, the low-viscosity compositions contain about 1% to about 10% of a viscosity-enhancing agent (e.g., a gelling component such as a polyoxyethylene-polyoxypropylene copolymer). In some embodiments, the low-viscosity compositions contain about 2% to about 10% of a viscosity-enhancing agent (e.g., a gelling component such as a polyoxyethylene-polyoxypropylene copolymer). In some embodiments, the low-viscosity compositions contain about 5% to about 10% of a viscosity-enhancing agent (e.g., a gelling component such as a polyoxyethylene-polyoxypropylene copolymer). In some embodiments, the low-viscosity compositions are substantially free of a viscosity-enhancing agent (e.g., a gelling component such as a polyoxyethylene-polyoxypropylene copolymer). In some embodiments, the low-viscosity ophthalmic pharmaceutical compositions described herein provide an apparent viscosity of about 100 cps to about 10,000 cps. In some embodiments, the low-viscosity ophthalmic pharmaceutical compositions described herein provide an apparent viscosity of about 500 cps to about 10,000 cps. In some embodiments, the low viscosity ophthalmic pharmaceutical compositions described herein provide an apparent viscosity of about 1000 cps to about 10,000 cps.

[0419] In some embodiments, the compositions described herein are viscous compositions at body temperature. In some embodiments, the viscous compositions contain from about 10% to about 25% of a viscosity enhancing agent (e.g., a gelling component such as a polyoxyethylene-polyoxypropylene copolymer). In some embodiments, the viscous compositions contain from about 14% to about 22% of a viscosity enhancing agent (e.g., a gelling component such as a polyoxyethylene-polyoxypropylene copolymer). In some embodiments, the viscous compositions contain from about 15% to about 21% of a viscosity enhancing agent (e.g., a gelling component such as a polyoxyethylene-polyoxypropylene copolymer). In some embodiments, the viscous compositions described herein provide an apparent viscosity of from about 100,000 cps to about 1,000,000 cps. In some embodiments, the viscous compositions described herein provide an apparent viscosity of from about 150,000 cps to about 500,000 cps. In some embodiments, the viscous compositions described herein provide an apparent viscosity of from about 250,000 cps to about 500,000 cps. In some such embodiments, the viscous compositions are liquid at room temperature and gel between room temperature and body temperature (e.g., up to about 42°C, including in individuals with severe fever). In some embodiments, the viscous compositions are administered as a monotherapy for the treatment of an ocular disease or condition described herein.

[0420] In some embodiments, the viscosity of the gel compositions provided herein is measured by any of the methods described. For example, in some embodiments, an LVDV-II+CP Cone Plate Viscometer and a Cone Spindle CPE-40 are used to calculate the viscosity of the gel compositions described herein. In other embodiments, a Brookfield (spindle and cup) viscometer is used to calculate the viscosity of the gel compositions described herein. In some embodiments, the viscosity ranges referred to herein are measured at room temperature. In other embodiments, the viscosity ranges referred to herein are measured at body temperature (e.g., the average body temperature of a healthy human).

[0421] Dose-to-dose consistency of gel / ointment A typical ophthalmic gel is packaged in an eye dropper bottle and administered as droplets. For example, a single administration (i.e., a single dose) of an ophthalmic gel may include one, two, three, or more drops administered to a patient's eye. Furthermore, a typical ophthalmic ointment is packaged in a tube or other compressible container with a dispensing nozzle that delivers strips of ointment. For example, a single administration (i.e., a single dose) of an ointment may include one strip or multiple strips administered to a patient's eye. In some embodiments, a single dose of the ophthalmic gel described herein is one drop of the gel composition from an eye dropper bottle. In some embodiments, a single dose of the ophthalmic ointment is one strip of the ointment composition dispensed through the nozzle of a dispensing tube.

[0422] In some cases, the present disclosure provides ophthalmic gel compositions that provide a uniform concentration per dose. In some instances, the uniform concentration per dose does not exhibit significant variations in drug content from one dose to another. In some instances, the uniform concentration per dose provides a consistent drug content from one dose to another.

[0423] In some cases, the present specification describes an eye ointment composition that provides a uniform concentration per dose.In some examples, the uniform concentration per dose does not exhibit significant variations in drug content from one dose to another.In some examples, the uniform concentration per dose provides a consistent drug content from one dose to another.

[0424] In some embodiments, the composition has a dose-to-dose ophthalmic agent concentration variation of less than 50%. In some embodiments, the composition has a dose-to-dose ophthalmic agent concentration variation of less than 40%. In some embodiments, the composition has a dose-to-dose ophthalmic agent concentration variation of less than 30%. In some embodiments, the composition has a dose-to-dose ophthalmic agent concentration variation of less than 20%. In some embodiments, the composition has a dose-to-dose ophthalmic agent concentration variation of less than 10%. In some embodiments, the composition has a dose-to-dose ophthalmic agent concentration variation of less than 5%.

[0425] In some embodiments, the concentration variation of the ophthalmic agent per dose is based on 10 consecutive doses. In some embodiments, the concentration variation of the ophthalmic agent per dose is based on 8 consecutive doses. In some embodiments, the concentration variation of the ophthalmic agent per dose is based on 5 consecutive doses. In some embodiments, the concentration variation of the ophthalmic agent per dose is based on 3 consecutive doses. In some embodiments, the concentration variation of the ophthalmic agent per dose is based on 2 consecutive doses.

[0426] A non-settling composition should not require shaking to uniformly distribute the drug. "No-shake" compositions are potentially advantageous over compositions requiring shaking for the simple reason that patient shaking behavior is a major source of variability in the amount of drug administered. It has been reported that patients often do not shake or forget to shake compositions requiring shaking before administering a dose, despite clear instructions to do so on the label. On the other hand, even patients who do shake the product generally cannot determine whether the strength and / or duration of shaking is adequate to achieve uniformity in the product. In some embodiments, the ophthalmic gel and ointment compositions described herein are "no-shake" compositions that maintain the dose-to-dose uniformity described herein.

[0427] To assess dose-to-dose uniformity, eye drop bottles or tubes containing the aqueous ophthalmic composition, ophthalmic gel composition, or ophthalmic ointment composition are stored upright for a minimum of 12 hours before the start of testing. To simulate the recommended dosing of these products, a predetermined number of drops or strips are dispensed from each commercial bottle or tube at predetermined time intervals over an extended period of time, or until no product remains in the commercial bottle or tube. All drops and strips are dispensed into tared glass bottles, 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.

[0428] Treatment method In one aspect, provided herein are methods for treating an ocular 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 repetitive sequence. In some embodiments, the repetitive sequence does not include multiple copies of GAA. In some embodiments, the repetitive sequence comprises repeats 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 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.

[0429] In embodiments 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 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).

[0430] In the embodiments of the treatment method provided herein, the double-stranded DNA binding moiety binds to double-stranded DNA non-covalently.In some embodiments, the double-stranded DNA binding moiety binds to double-stranded DNA using electrostatic force such as hydrogen bond or van der Waals force.In some embodiments, the double-stranded DNA binding moiety does not comprise polynucleotide.In some embodiments, the double-stranded DNA binding moiety does not comprise polypeptide.

[0431] In embodiments of the treatment methods provided herein, 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, less than about 7.5 kDa, less than about 5 kDa, or less than about 2.5 kDa.

[0432] 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 connected to the transcriptional modulator-binding moiety by a linker.

[0433] In aspects of the treatment 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 human.

[0434] In embodiments of the methods of treatment 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 comprises a tonicity adjusting agent, e.g., sodium chloride, a buffer, a stabilizer, an antioxidant, a viscosity enhancing agent, a solubilizing agent, a penetration enhancer, and / or a preservative. In some embodiments, the 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.

[0435] In various embodiments, administration of the composition to the eye results in minimal systemic exposure. In some embodiments, local administration of the composition to the eye (e.g., ocular administration) results in the 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 penetrates the internal cells and tissues of the eye.

[0436] In another aspect, provided herein are methods for treating an ocular disorder 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 the composition is suitable for administration to a human eye. In some embodiments, the double-stranded DNA-binding moiety does not bind to a repetitive sequence containing multiple copies of GAA. In some embodiments, the double-stranded DNA-binding moiety binds to a repetitive sequence containing 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 contains multiple copies of CTG or CAG. In some embodiments, the repetitive sequence contains at least four nucleotides. In some embodiments, the repetitive sequence is not a triplet repeat.

[0437] In some embodiments of the treatment method using 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.

[0438] In embodiments of the treatment methods provided herein, the double-stranded DNA-binding moiety that does not contain nucleotides or amino acids 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 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).

[0439] In the embodiment of the treatment method provided herein, the double-stranded DNA binding moiety does not contain nucleotide or amino acid and binds to double-stranded DNA non-covalently.In some embodiments, the double-stranded DNA binding moiety binds to double-stranded DNA using electrostatic force such as hydrogen bond or van der Waals force.In some embodiments, the double-stranded DNA binding moiety does not contain polynucleotide.In some embodiments, the double-stranded DNA binding moiety does not contain polypeptide.

[0440] In embodiments of the treatment methods provided herein, the double-stranded DNA binding moiety that does not contain nucleotides or amino acids 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, less than about 7.5 kDa, less than about 5 kDa, or less than about 2.5 kDa.

[0441] In some embodiments, the double-stranded DNA-binding moiety does not contain nucleotides or amino acids and 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 connected to the transcriptional modulator-binding moiety by a linker.

[0442] In the embodiment of the treatment method in which the double-stranded DNA binding moiety does not comprise nucleotide or amino acid provided herein, the individual is a mammal.In some embodiments, the individual is a mouse, a rat, a cat, a dog, a rabbit, a horse, a cow, a pig or a human.In some embodiments, the individual is a human.

[0443] In embodiments of the methods of treatment provided herein in which the double-stranded DNA-binding moiety does not comprise a nucleotide or an amino acid, the double-stranded DNA-binding moiety is formulated in a composition comprising an excipient suitable for ocular administration. In some embodiments, the excipient comprises a tonicity adjuster, e.g., sodium chloride, a buffer, a stabilizer, an antioxidant, a viscosity enhancer, a solubilizer, a penetration enhancer, and / or a preservative. In some embodiments, the 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.

[0444] In various embodiments, administration of a composition in which the double-stranded DNA-binding moiety 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., ocular administration) results in the double-stranded DNA-binding moiety penetrating 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 penetrates into the internal cells and tissues of the eye.

[0445] In a further aspect, provided herein is a method 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 contain a nucleotide or amino acid.

[0446] In some embodiments of the method for treating an eye disorder, the eye disorder is a genetic disease, and the double-stranded DNA binding moiety does not bind to a repetitive sequence containing multiple copies of GAA. In some embodiments, the double-stranded DNA binding moiety binds to a repetitive sequence containing 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 contains multiple copies of CTG or CAG. In some embodiments, the repetitive sequence contains at least four nucleotides. In some embodiments, the repetitive sequence is not a triplet repeat. In some embodiments, the double-stranded DNA binding moiety binds to the minor groove of double-stranded DNA.

[0447] In an embodiment of the method for treating an ocular 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 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).

[0448] In the embodiments of the method for treating genetic eye disorders provided herein, the double-stranded DNA binding moiety binds to double-stranded DNA non-covalently.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.

[0449] In the embodiments of the methods for treating genetic eye disorders provided herein, 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, less than about 7.5 kDa, less than about 5 kDa, or less than about 2.5 kDa.

[0450] In some embodiments of the method for treating an eye disorder that is a genetic disease, 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 connected to the transcriptional modulator-binding moiety by a linker.

[0451] In the aspects of the methods for treating genetic eye disorders 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 human.

[0452] In embodiments of the methods for treating genetic eye disorders 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 comprises a tonicity adjusting agent, e.g., sodium chloride, a buffer, a stabilizer, an antioxidant, a viscosity enhancing agent, a solubilizing agent, a penetration enhancer, and / or a preservative. In some embodiments, the 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.

[0453] In various embodiments of the method for treating genetic eye disorders, administering the composition to the eye results in minimal systemic exposure. In some embodiments, local administration of the composition to the eye (e.g., ocular administration) results in the 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 penetrates the internal cells and tissues of the eye.

[0454] In various embodiments of the treatment methods provided herein, tissue biodistribution can be evaluated in single and multiple dose ocular administration tests carried out in experimental models such as rats, rabbits and dogs.In some cases, after ocular administration, the level of double-stranded DNA binding agent in ocular tissues consistently shows a gradient of exposure, with the highest concentration of double-stranded DNA binding agent measured in the outermost ocular structure closest to the application site, and then decreases to the innermost corneal endothelial layer, which is the therapeutic target tissue of double-stranded DNA binding agent.

[0455] In some cases, the double-stranded DNA binding agent is administered at a concentration equal to or greater than 3-20 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, after about 14 days of repeated doses, e.g., for 1-28 days, 7-28 days, 14-28 days, 21-28 days, 1-21 days, 7-21 days, 14-21 days, 1-14 days, 7-14 days, or 1-7 days. After days (once or twice daily), the concentration is detectable in the corneal endothelium for up to 24 hours, e.g., 2-24 hours, 6-24 hours, 12-24 hours, 18-24 hours, 2-18 hours, 6-18 hours, 12-18 hours, 2-12 hours, 6-12 hours, or 2-6 hours, which is a concentration predicted to reduce the amount of nuclear foci and symptoms, such as cases of spliceopathy, in corneal endothelial cells (CECs) of some affected individuals. In one embodiment, after the final ophthalmic dose, for example, 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 times, 20 times, 30 times, 40 times, 50 times, 60 times, 80 times, 100 times, or 120 times higher than the lowest or highest predicted effective concentration. In some cases, the effective concentration is achieved when the double-stranded DNA binding agent is administered once daily for several days, weeks, or months using a clinically relevant administration paradigm. In some cases, the effective concentration is achieved when the double-stranded DNA binding agent is administered twice daily for several days, weeks, or months using a clinically relevant administration paradigm. In some cases, results from distribution studies utilizing water formulations, such as acidified water formulations, demonstrate that ophthalmic administration results in concentrations of double-stranded DNA binding agent in the target endothelium that exceed concentrations predicted to be biologically relevant for approximately 24 hours after administration.

[0456] In various embodiments of the methods of treatment provided herein, an aqueous formulation, e.g., an acidified aqueous formulation, optionally containing 1-10% mannitol, e.g., 2-10%, 2-8%, 2-6%, 2-4%, 4-10%, 4-8%, 4-6%, 6-10%, 6-8%, or 8-10% mannitol, is selected as the ophthalmic formulation for the double-stranded DNA binding agent. In some cases, the double-stranded DNA binding agent formulated in this manner is administered at an ophthalmic dose regimen (e.g., 4 drops of 50 μL twice daily in both eyes, e.g., 1, 2, 3, 4, 5, or 6 drops of 10 μL, 20 μL, 30 μL, 40 μL, 50 μL, 60 μL, 70 μL, 80 μL, 90 μL, or 100 μL twice daily per eye) and concentration (0.6% to 1%, e.g., 0.1%, 0.2%, 0.3%, 0.4%, Utilizing 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%), topical administration has been well tolerated after daily administration for approximately 7 days, e.g., 1-14 days, 1-10 days, 1-8 days, 1-6 days, 1-4 days, 1-2 days, 2-14 days, 2-10 days, 2-8 days, 2-6 days, 2-4 days, 4-14 days, 4-10 days, 4-8 days, 4-6 days, 6-14 days, 6-10 days, 6-8 days, 8-14 days, 8-10 days, or 10-14 days. In some cases, ocular target tissue levels are equivalent to those observed in earlier studies using acidified water alone. In some cases, peak exposure of the double-stranded DNA binding agent is observed in the corneal endothelium within the first hour after topical instillation, e.g., within 20 minutes, 40 minutes, or 1 hour after treatment. A decrease in the level of double-stranded DNA binding agent in the corneal endothelium over the next 2 to 24 hours, e.g., 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, or 24 hours, to about 3 to 40 nM, e.g., 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, suggests diffusion and / or transport across the cornea.In some cases, the double-stranded DNA binding agent is detectable in the aqueous humor for up to 12 to 48 hours after the last administration, e.g., 12, 16, 20, 24, 28, 32, 36, 40, 44, or 48 hours. In some cases, the entire volume of aqueous humor is replenished approximately every 1 to 4 hours, e.g., every 1, 2, 3, or 4 hours, in research animals such as rabbits; therefore, measurable levels of double-stranded DNA binding agent in the aqueous humor at later time points suggest sustained release of the double-stranded DNA binding agent from the cornea to the posterior segment of the eye.

[0457] [Table 1-1]

[0458] [Table 1-2]

[0459] [Table 1-3]

[0460] [Table 1-4] [Example]

[0461] The following examples are given for the purpose of illustrating various embodiments of the present invention and are not intended to limit the invention in any way. The examples, along with the methods described herein, are representative of preferred embodiments, are exemplary, and are not intended to limit the scope of the invention. Variations thereof and other uses encompassed within the spirit of the invention as defined by the scope of the claims will occur to those skilled in the art.

[0462] Example 1: Determining the solubility of compounds in water To determine the solubility of candidate compounds, in situ salt formation with aqueous HCl was used to determine the solubility of the compounds at a concentration of 10 mg / ml. First, 0.5 ml of sterile water was added to 10 mg of dry compound, vortexed, and briefly centrifuged. Next, 0.5 equivalents of 0.025 M HCl was added to the mixture, followed by a total volume of 1 ml of water. The mixture was continuously stirred for 0.5 to 24 hours. If a clear solution was obtained, the pH was determined using a microelectrode. If the solution was not clear, additional water and / or HCl was added with continuous stirring and vortexing until the compound was completely dissolved or determined to be resistant to dissolution. The minimum concentration tested was 1 mg / ml. The pH was evaluated at the end of the experiment. The experimental results are shown in Table 2.

[0463] [Table 2]

[0464] Example 2: Determining compound activity using the in vitro foci assay

[0465] Immortalized Fuchs corneal endothelial dystrophy cells (F35T) were seeded at 5,000 cells per well in a 96-well plate with a black side and clear bottom 16 hours before compound treatment. Compounds were dissolved in DMSO to 1 mM and diluted in F35T cell culture medium to a final concentration of 3,000–1 nM. The diluted compounds were added to F35T cells and incubated for 48 hours.

[0466] After treatment, 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, cells were washed twice with PBS and then hydrated with 30% formamide + 2x SSC buffer for 10 minutes. Fluorescence in situ hybridization with (CAG)10-Cy3 oligonucleotide was used to label nuclear foci. Images were collected, and automated image analysis was performed on a BioTEK Cytation 5. Experimental data are shown in Table 3.

[0467] [Table 3]

[0468] Example 3: Evaluation of ocular penetration of compounds formulated in water Freshly enucleated bovine eyes (Nebraska Scientific) were treated with a previously described compound (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)). Each eye was rinsed with 10 mL of sterile 0.9% saline and then placed in the bottom of a 600 mL glass beaker (Pyrex). A borosilicate glass powder funnel (Eisco Labs) with a 26 mm diameter base was gently placed over the eye with the cornea centered in the cylindrical opening, creating a sealed dosing chamber into which the solution was applied. The eyes were equilibrated in sterile 0.9% saline at room temperature for 20 minutes and then decanted. 500 μL of 0.1–0.3% GeneTAC compound was applied to the cornea, followed by 167 μL of sterile water to mimic the instillation of 30 μL of eye drops into 10 μL of tear fluid. The compound was incubated at room temperature for 30 minutes, after which the funnel was removed. The eye was then rinsed twice with 10 mL of 5.2% mannitol in water to remove residual compound. Aqueous humor was collected using a 28-gauge needle and flash-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 flash-frozen in liquid nitrogen. Liquid chromatography-mass spectrometry was used to quantify compound abundance in the aqueous humor, anterior cornea, and DM / CE. The data are shown in Table 4 below. The data are also presented in Figure 1, which shows that compound 201 reaches concentrations above the IC50 for preventing foci formation and above the IC50 for preventing aberrant splicing events in the corneal epithelium and stroma, as well as Descemet's membrane and endothelium.

[0469] [Table 4]

[0470] While preferred embodiments of the present invention have been shown and described herein, it will be obvious to those skilled in the art that such embodiments are provided by way of example only. Numerous variations, changes, and substitutions will occur to those skilled in the art without departing from the invention. It is understood that various alternatives to the embodiments described herein may be employed. The following claims define the scope of the invention, and it is intended that methods and structures within the scope of the claims and their equivalents be covered thereby.

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 repetitive sequences containing multiple copies of GAA, and the composition is suitable for administration to the human eye.

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

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

4. 1. A composition comprising a double-stranded DNA binding moiety that binds to a repeat sequence containing 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 of any one of claims 1 to 5, wherein the double-stranded DNA binding moiety binds to a repetitive sequence containing multiple copies of CTG or CAG.

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

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

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

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

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

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

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

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

17. The composition of claim 16 , wherein the double-stranded DNA binding portion is connected to the transcription modulator binding portion by a linker.

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

19. The composition of any one of claims 1 to 18, further comprising an excipient.

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

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

22. 1. A method for treating an eye disorder in a human individual in need thereof, the method comprising administering a double-stranded deoxyribonucleic acid (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 repetitive sequences containing multiple copies of GAA.

23. 1. A method of treating an ocular disorder in a human individual in need thereof, said method comprising administering a double-stranded DNA binding moiety, wherein said ocular disorder is a genetic disease.

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

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

26. 1. A method for treating an ocular disorder in a human 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.

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

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

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

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

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

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

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

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

38. 38. The method of claim 37, wherein the double-stranded DNA binding moiety is connected to the transcription modulator binding moiety by a linker.

39. 39. The method of any one of claims 22 or 25-38, wherein the double-stranded DNA binding moiety is formulated in an excipient suitable for administration to the eye.

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

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

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