Methods and compounds for identifying glycosyltransferase inhibitors

• US 10,106,833 B2
• Filed: 03/13/2013
• Issued: 10/23/2018
• Est. Priority Date: 04/06/2012
• Status: Active Grant

• Current Assignee: President and Fellows of Harvard College (Harvard Corporation)
• Original Assignee: President and Fellows of Harvard College (Harvard Corporation)
• Inventors: Kahne, Daniel Evan, Kahne, Suzanne Walker, Gampe, Christian M., Tsukamoto, Hirokazu
• Primary Examiner(s): Olson, Eric
• Application Number:
  US14/390,857
• Publication Number:
  US 20150079618A1
• Time in Patent Office: 2,050 Days
• Field of Search: 536 168, 536 41, 536 551, 435194, 435 74, 514 25, 514 54, 514 61
• CPC Class Codes: C07H 11/04 Phosphates; Phosphites; Polyphosphates
• CPC Class Codes: C07H 13/12 by acids having the group -X-C(=X)-X-, or halides thereof, in which each X means nitrogen, oxygen, sulfur, selenium or tellurium, e.g. carbonic acid, carbamic acid
• CPC Class Codes: C07H 15/26 Acyclic or carbocyclic radicals, substituted by hetero rings
• CPC Class Codes: C12Q 1/48 involving transferase
• CPC Class Codes: G01N 2333/91091 Glycosyltransferases (2.4)
• CPC Class Codes: G01N 2500/04 Screening involving studying the effect of compounds C directly on molecule A (e.g. C are potential ligands for a receptor A, or potential substrates for an enzyme A)
• CPC Class Codes: G01N 2500/20 cell-free systems

Abstract

The present invention provides moenomycin-based probe compounds of Formula (I) for use in screening inhibitors of bacterial glycosyltransferases. The present invention also provides bacterial glycosyltransferase screening assays using compounds of Formula (I).

Claims

1. A compound of Formula (I):

2. A compound of Formula (I):

3. The compound of claim 2, wherein R^YY is —

OH or —

OR^XX, and R^XX is a group of formula;

4. The compound of claim 2, wherein R^XX is a group of formula:

5. The compound of claim 2, wherein R⁶ is —

CH₂OH or —

CH₂OR^CX;

wherein R^CX is a carbohydrate moiety.

6. The compound of claim 2, wherein G is

7. The compound of claim 2, wherein R^P is selected from the group consisting of Alexa Fluor 350, Alexa Fluor 488, Alexa Fluor 532, Alexa Fluor 546, Alexa Fluor 568, Alexa Fluor 594, Alexa Fluor 633, Alexa Fluor 660 and Alexa Fluor 680, AMCA, AMCA-S, BODIPY FL, BODIPY R6G, BODIPY TMR, BODIPY TR, BODIPY 493/503, BODIPY 530/550, BODIPY 558/568, BODIPY 564/570, BODIPY 576/589, BODIPY 581/591, BODIPY 630/650, BODIPY 650/665, aminomethylcoumarin, carbocyanine, carboxyrhodamine 6G, carboxy-X-rhodamine (ROX), Cascade Blue, Cascade Yellow, coumarin, coumarin 343, cyanine dyes, dansyl, dapoxyl, dialkylaminocoumarin, 4′

,5′

-dichloro-2′

,7′

-dimethoxyfluorescein, DM-NERF, eosin, erythrosin, fluorescein, FAM, hydroxycoumarin, IRD40, IRD 700, IRD 800, 6-carboxy-4′

,5′

-dichloro-2′

,7′

-dimethoxyfluorescein (6-JOE), lissamine rhodamine B, Marina Blue, merocyanine, methoxycoumarin, naphthofluorescein, Oregon Green 488, Oregon Green 500, Oregon Green 514, oxonol dyes, Pacific Blue, phycoerythrin, PyMPO, pyrene, rhodamine B, rhodamine 6G, rhodamine green, rhodamine red, rhodol green, styryl dyes, 2′

,4′

,5′

,7′

-tetrabromosulfone-fluorescein, tetramethyl-rhodamine (TMR), carboxytetramethylrhodamine (TAMRA), Texas Red, Texas Red-X, 5(6)-carboxyfluorescein, 2,7-dichlorofluorescein, N,N-bis(2,4,6-trimethylphenyl)-3,4,9,10-perylenebis(dicarboximide), HPTS, ethyl eosin, DY-490XL MegaStokes, DY-485XL MegaStokes, Adirondack Green 520, ATTO 465, ATTO 488, ATTO 495, Y0Y0-1,5-FAM, BCECF, dichlorofluorescein, rhodamine 110, rhodamine 123, YO-PRO-I, SYTOX Green, Sodium Green, SYBR Green I, Alexa Fluor 500, FITC, Fluo-3, Fluo-4, fluoro-emerald, YoYo-I ssDNA, YoYo-I dsDNA, YoYo-I, SYTO RNASelect, Diversa Green-FP, Dragon Green, EvaGreen, Surf Green EX, Spectrum Green, Spectrum Red, NeuroTrace 500525, NBD-X, MitoTracker Green FM, LysoTracker Green DND-26, CBQCA, PA-GFP (post-activation), WEGFP (post-activation), FLASH-CCXXCC, Azami Green monomeric, Azami Green, green fluorescent protein (GFP), EGFP, Kaede Green, 7-benzylamino-4-nitrobenz-2-oxa-1,3-diazole, Bexl, doxorubicin, Lumio Green, and SuperGlo GFP.

8. The compound of claim 2 of formula:

9. The compound of claim 2, wherein:

R¹ is —

C(O)NHR⁸, —

CH₂OR⁹, or —

C(O)OR⁹;

one of R² and R³ is hydrogen, and the other is —

OR⁹;

R⁴ is —

W—

R^4a;

R⁵ is —

NHR⁸;

R⁶ is —

CH₃, —

CH₂OR⁹, or —

CH₂OR^CX;

R⁷ is —

OR⁹ or —

N(R⁸)₂;

R¹¹ is —

CH₃, —

C(O)NHR⁸, —

CH₂OR⁹, or —

C(O)OR⁹;

one of R¹² and R¹³ is hydrogen, and the other is —

OR⁹;

R¹⁴ is —

NHR⁸; and

one of R^19a and R^19b is hydrogen, and the other is —

OR⁹.

10. The compound of claim 3, wherein R¹¹ is —

CH₂OH.

11. The compound of claim 2, wherein R^P is selected from the group consisting of fluorescein, naphthofluorescein, 4′

,5′

-dichloro-2′

,7′

-dimethoxy-fluorescein, 2′

,4′

,5′

,7′

-tetrabromosulfone-fluorescein, 2,7-dichlorofluorescein, 6-carboxy-4′

,5′

-dichloro-2′

,7′

-dimethoxyfluorescein, and 5(6)-carboxyfluorescein.

12. The compound of claim 11, wherein R^P is fluorescein.

13. The compound of claim 3, wherein R¹¹ is —

CH₃.

14. The compound of claim 2, wherein R¹² is hydrogen;

and R¹³ is —

OH.

15. The compound of claim 2, wherein G is of Formula (a).

16. The compound of claim 2, wherein G is of Formula (b).

17. A compound of Formula (I):