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EP-4735534-A2 - BENZOTHIAZOLE DYES

EP4735534A2EP 4735534 A2EP4735534 A2EP 4735534A2EP-4735534-A2

Abstract

Provided herein are compounds having aqueous solubility, stability, and fluorescence, as well as methods of detecting the presence or absence of an analyte in a test sample using said compounds.

Inventors

  • Kamber, David N.
  • Nani, Roger R.
  • HANSEN, WILLIAM A.

Assignees

  • Detect, Inc.

Dates

Publication Date
20260506
Application Date
20240628

Claims (20)

  1. CLAIMS 1. A compound of Formula (I): or a salt thereof, wherein: n is 1-4; m is 0-4; X is oxygen or sulfur; R 1 is hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, or substituted or unsubstituted heteroaryl; R 2 is, independently for each occurrence, halo, -OR 5 , -SR 5 , -N(R 5 ) 2 , substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted carbocyclyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; wherein two adjacent R 2 groups may combine to form a fused substituted or unsubstituted aryl, a fused substituted or unsubstituted heteroaryl, a fused substituted or unsubstituted carbocyclyl, or a fused substituted or unsubstituted heterocyclyl; R 3 is substituted or unsubstituted alkyl; R 4 is, independently for each occurrence, halo, -NO2, -OR 6 , -SR 6 , -N(R 6 )2, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted carbocyclyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; R 5 is, independently for each occurrence, hydrogen, substituted or unsubstituted alkyl, nitrogen protecting group when attached to a nitrogen, oxygen protecting group when attached to an oxygen, or a sulfur protecting group when attached to a sulfur, or when R 2 is - N(R 5 )2, two instances of R 5 may be joined to form a substituted or unsubstituted heterocyclyl or a substituted or unsubstituted heteroaryl; and R 6 is, independently for each occurrence, hydrogen, substituted or unsubstituted alkyl, nitrogen protecting group when attached to a nitrogen, oxygen protecting group when attached to an oxygen, or a sulfur protecting group when attached to a sulfur. 71/82 12590649.1
  2. 2. The compound of claim 1, or a salt thereof, wherein X is sulfur.
  3. 3. The compound of claim 1 or 2, or a salt thereof, wherein R 1 is hydrogen, substituted or unsubstituted alkyl, or substituted or unsubstituted heteroalkyl.
  4. 4. The compound of any one of claims 1-3, or a salt thereof, wherein R 1 is not substituted or unsubstituted heteroaryl.
  5. 5. The compound of any one of claims 1-4, or a salt thereof, wherein R 1 is substituted or unsubstituted alkyl.
  6. 6. The compound of any one of claims 1-5, of a salt thereof, wherein R 1 is substituted or unsubstituted C1-3 alkyl.
  7. 7. The compound of claim any one of claims 1-5, or a salt thereof, wherein R 1 is unsubstituted alkyl.
  8. 8. The compound of any one of claims 1-7, of a salt thereof, wherein R 1 is methyl.
  9. 9. The compound of any one of claims 1-6, or a salt thereof, wherein R 1 is alkyl substituted with an amine.
  10. 10. The compound of any one of claims 1-6, or a salt thereof, wherein R 1 is alkyl substituted with a quaternary ammonium.
  11. 11. The compound of any one of claims 1-10, or a salt thereof, wherein n is 1 or 2.
  12. 12. The compound of any one of claims 1-11, of the Formula (I-a) or (I-b): 72/82 12590649.1 or a salt thereof.
  13. 13. The compound of any one of claims 1-12, or a salt thereof, wherein at least one R 2 is substituted or unsubstituted heterocyclyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, or -N(R 5 ) 2 .
  14. 14. The compound of any one of claims 1-12, or a salt thereof, wherein two adjacent R 2 groups combine to form a fused substituted or unsubstituted aryl.
  15. 15. The compound of any one of claims 1-14, or a salt thereof, wherein at least one R 2 is selected from: 73/82 12590649.1
  16. 16. The compound of any one of claims 1-10, or a salt thereof, wherein at least one R 2 is two adjacent R 2 groups combine to form .
  17. 17. The compound of any one of claims 1-16, or a salt thereof, wherein at least one R 2 is .
  18. 18. The compound of any one of claims 1-17, or a salt thereof, wherein R 3 is unsubstituted alkyl.
  19. 19. The compound of any one of claims 1-18, of Formula (I-c): or a salt thereof.
  20. 20. The compound of claim 1, of formula (I-d): 74/82 12590649.1 or a salt thereof, wherein: p is 1-4; and R 7 is -CH 3 or a quaternary ammonium group.

Description

BENZOTHIAZOLE DYES CROSS-REFERENCE TO RELATED APPLICATIONS [001] The present application claims the benefit of and priority under 35 U.S.C. § 119(e) to U.S. Provisional Application No.63/511,610 filed June 30, 2023, titled BENZOTHIAZOLE DYES, the contents of which are incorporated herein by reference in their entirety. BACKGROUND [002] Dyes are commonly used in the biotechnological and biomedical research fields for the detection of molecules of interest, such as nucleic acids. The detection and quantification of nucleic acids, particularly DNA, using fluorescent dyes in biological test samples is an important tool in the diagnosis of various genetic or pathological conditions. As more sophisticated diagnostic and testing platforms are developed, there is an increased need for dye compounds having properties amenable to the new mechanical and chemical requirements of the platforms. SUMMARY OF THE INVENTION [003] In one aspect, the present disclosure provides a compound of Formula (I): or a salt thereof, wherein: n is 1-4; m is 0-4; X is oxygen or sulfur; R1 is hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, or substituted or unsubstituted heteroaryl; R2 is, independently for each occurrence, halo, -OR5, -SR5, -N(R5)2, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted carbocyclyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; wherein two adjacent R2 groups may combine to form a fused substituted or unsubstituted aryl, a fused substituted or unsubstituted heteroaryl, a 1/82 12590649.1 fused substituted or unsubstituted carbocyclyl, or a fused substituted or unsubstituted heterocyclyl; R3 is substituted or unsubstituted alkyl; R4 is, independently for each occurrence, halo, -NO2, -OR6, -SR6, -N(R6)2, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted carbocyclyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; R5 is, independently for each occurrence, hydrogen, substituted or unsubstituted alkyl, nitrogen protecting group when attached to a nitrogen, oxygen protecting group when attached to an oxygen, or a sulfur protecting group when attached to a sulfur, or when R2 is - N(R5)2, two instances of R5 may be joined to form a substituted or unsubstituted heterocyclyl or a substituted or unsubstituted heteroaryl; and R6 is, independently for each occurrence, hydrogen, substituted or unsubstituted alkyl, nitrogen protecting group when attached to a nitrogen, oxygen protecting group when attached to an oxygen, or a sulfur protecting group when attached to a sulfur. [004] In another aspect, the present disclosure provides a composition comprising a compound described herein, or a salt thereof. [005] In another aspect, the present disclosure provides a method of detecting the presence or absence of an analyte in a test sample, comprising contacting the test sample with a compound of the present disclosure in the presence of an energy (E1) and detecting emission of an energy (E2) from the sample. [006] In another aspect, the present disclosure provides a kit comprising: a compound disclosed herein; or a composition of the present disclosure; and instructions for use. BRIEF DESCRIPTION OF THE DRAWINGS [007] The accompanying drawings, which constitute a part of this specification, illustrate several embodiments of the invention and together with the description, provide non-limiting examples of the invention. [008] FIG.1A shows time to positive (Tp) data for Chlamydia trachomatis (RS2340-16) qLAMP assay with titration of dyes (4.0µM, 8.0µM, 10.0µM, 16.0µM), left = clean positive control, right = clean negative control. FIG.1B shows Tp data for Neisseria gonorrhoeae trachomatis (WX60) qLAMP assay with titration of dyes (4.0µM, 8.0µM, 10.0µM, 16.0µM), left = clean positive control, right = clean negative control. 2/82 12590649.1 [009] FIG.2 shows Tp data for Neisseria gonorrhoeae trachomatis (AQ10) qLAMP assay with titration of dyes (4.0µM, 8.0µM, 10.0µM, 16.0µM), left = clean positive control, right = clean negative control. [010] FIG.3A shows time to positive (RES) results for qLAMP assays using Flub, H1N1, SC2, and NG2 primer sets with compound 2 at varying concentrations (NTC = non-template control). FIG.3B shows Tp (RES) results for qLAMP assays using Flub, H1N1, SC2, and NG2 primer sets with compound 1 at varying concentrations (NTC = non-template control). [011] FIG.4 shows RES data for Neisseria gonorrhoeae trachomatis (AQ10) qLAMP assay with compound 3 at varying temperatures. [012] FIG.5 shows stability data for compound 3 under accelerated stability conditions (40 °C, lyophilized samples) over 1-3 months. [013] FIG.6A-F show absorption/emission spectra of dyes with double stranded DNA (dsDNA). FIG.6A shows a