Small molecule printing

a technology of small molecules and printing, applied in the field of small molecules printing, can solve the problems of slow serial process and limited to only a few iterations

Inactive Publication Date: 2005-05-05
PRESIDENT & FELLOWS OF HARVARD COLLEGE
View PDF66 Cites 8 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0016]“Small Molecule”: As used herein, the term “small molecule” refers to a non-peptidic, non-oligomeric organic compound either synthesized in the laboratory or found in nature. Small molecules, as used herein, can refer to compounds that are “natural product-like”, however, the term “small molecule” is not limited to “natural product-like” compounds. Rather, a small molecule is typically characterized in that it contains several carbon-carbon bonds, and has a molecular weight of less than 1500, although this characterization is not intended to be limiting for the purposes of the present invention. Examples of “small molecules” that occur in nature include, but are not limited to, taxol, dynemicin, and rapamycin. Examples of “small molecules” that are synthesized in the laboratory include, but are not limited to, compounds described in Tan et al., (“Stereoselective Synthesis of over Two Million Compounds Having Structural Features Both Reminiscent of Natural Products and Compatible with Miniaturized Cell-Based Assays”J. Am. Chem. Soc. 1998, 120, 8565) and pending application Ser. No. 08 / 951,930 “Synthesis of Combinatorial Libraries of Compounds Reminiscent of Natural Products”, the entire contents of which are incorporated herein by reference. In certain other preferred embodiments, natural-product-like small molecules are utilized.

Problems solved by technology

Despite the proven utility of this approach, it is limited by the small number of proteins that can be screened efficiently.
In principle, the beads can be stripped of one protein and reprobed with another; however, this serial process is slow and limited to only a few iterations.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Small molecule printing
  • Small molecule printing
  • Small molecule printing

Examples

Experimental program
Comparison scheme
Effect test

example 1

Small Molecule Printing using Michael Addition

[0054] In order to demonstrate the utility of small molecule printing as a technique identifying small molecule-protein interactions, three unrelated molecules were chosen for which specific protein receptors are available. Compound 1 (FIG. 6, R═OH) is the vitamin biotin, which is recognized by the bacterial protein streptavidin (Chaiet et al., Arch. Biochem. Biophys. 1964, 106, 1; incorporated herein by reference). Compound 2 (R═OH) is a derivative of the steroid digoxigenin and is recognized by the mouse monoclonal antibody DI-22 (Sigma). Finally, compound 3 (R═OH) is a synthetic pipecolyl α-ketoamide, which was designed to be recognized by the human immunophilin FKBP12 (Holt et al., J. Am. Chem. Soc. 1993, 115, 9925; incorporated herein by reference). Each of these compounds was attached to 400-450 μm diameter polystyrene beads (estimated capacity of 20 nmol per bead) via a 6-aminocaproic acid linker and either 4-methoxytrityl-protec...

example 2

Small Molecule Printing Using Silylation Reaction

[0058] Standard glass slides were activated for selective reaction with alcohols (FIG. 9). Microscopic slides were first treated with a H2SO4 / H2O2 solution (“piranha”) for 16 hours at room temperature. After extensive washing with water, the slides were treated with thionyl chloride and a catalytic amount of DMF in THF for 4 hours at room temperature. Surface characterization by x-ray photoelectron spectroscopy (XPS) confirmed the presence of chlorine on the slide (Strother et al., J. Am. Chem. Soc., 2000, 122, 1205-1209; incorporated herein by reference). To test the ability of these chlorinated slides to capture alcohols released from synthesis beads, we initially used three alcohol-containing small molecules and a bead linker reagent developed for chemical genetic applications of diversity-oriented synthesis.

[0059] Primary alcohol derivatives of a synthetic α-ketoamide (Holt et al., J. Am. Chem. Soc. 1993, 115, 9925-9938; incorpo...

example 3

Fabrication of Custom Slide Reaction Vessels

[0062] In an effort to minimize reagent volume during the chemical treatment of glass microscope slides, we designed and fabricated custom slide-sized reaction vessels that enable the uniform application of ˜1.4 mL solution to one face of a 2.5 cm×7.5 cm slide. First, a master template mold was cut from a block of Delhran plastic according to the blueprint shown in FIG. 13. The slide-sized reaction vessels were prepared by casting degassed polydimethysiloxane (PDMS, Sylgard Kit 184, Dow corning, Midland, Mich.) prepolymer around the master template in a polystyrene OmniTray (Nalge Nunc International, Naperville, Ill.). After curing for four hours at 65° C., the polymer was peeled away from the master to give the finished product (FIG. 14).

[0063] To use the vessels, slides were placed face-down as illustrated below and reagent was injected under the slides with a P1000 Pipetman (FIG. 15).

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

PropertyMeasurementUnit
distanceaaaaaaaaaa
diameteraaaaaaaaaa
volumeaaaaaaaaaa
Login to view more

Abstract

The present invention provides compositions and methods to facilitate the identification of compounds that are capable of interacting with a biological macromolecule of interest. In one aspect, a composition is provided that comprises an array of one or more types of chemical compounds attached to a solid support, wherein the density of the array of compounds is at least 1000 spots per cm2. In particularly preferred embodiments, these compounds are attached to the solid support through a covalent interaction. In general, these inventive arrays are generated by: (1) providing a solid support, wherein said solid support is functionalized with a selected chemical moiety capable of interacting with a desired chemical compound to form an attachment; (2) providing one or more solutions of one or more types of compounds to be attached to the solid support; and (3) delivering said one or more types of compounds to the solid support, whereby an array is formed and the array of compounds has a density of at least 1000 spots per cm2. In another aspect, the present invention provides methods for utilizing these arrays to identify small molecule partners for biological macromolecules of interest comprising: (1) providing an array of compounds, wherein the array has a density of at least 1000 spots per cm2; (2) contacting the array with one of more types of biological macromolecules of interest; and (3) determining the interaction of specific small molecule-biological macromolecule partners.

Description

RELATED APPLICATIONS [0001] The present application claims priority to provisional application U.S. Ser. No. 60 / 133,595, filed May 11, 1999, which is incorporated herein by reference in its entirety.BACKGROUND OF THE INVENTION [0002] The ability to identify small molecule ligands for any protein of interest has far-reaching implications, both for the elucidation of protein function and for the development of novel pharmaceuticals. With the introduction of split-pool strategies for synthesis (Furka et al., Int. J. Pept. Protein Res. 1991, 37,487; Lam et al., Nature 1991, 354, 82; each of which is incorporated herein by reference) and the development of appropriate tagging technologies (Nestler et al., J. Org. Chem. 1994, 59, 4723; incorporated herein by reference), chemists are now able to prepare large collections of natural product-like compounds immobilized on polymeric synthesis beads (Tan et al., J. Am. Chem. Soc. 1998, 120, 8565; incorporated herein by reference). These librari...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Applications(United States)
IPC IPC(8): B01J19/00C07B61/00C07C311/49C40B30/04C40B40/04C40B50/14C40B60/14G01N33/543
CPCB01J19/0046Y10S436/809B01J2219/00454B01J2219/00459B01J2219/005B01J2219/00533B01J2219/00605B01J2219/0061B01J2219/00612B01J2219/00626B01J2219/00637B01J2219/0072C07B2200/11C07C311/49C40B30/04C40B40/04C40B50/14C40B60/14G01N33/54353G01N31/00B01J2219/00387
Inventor SCHREIBER, STUART L.MACBEATH, GAVINKOEHLER, ANGELAHERGENROTHER, PAULDEPEW, KRISTOPHER
Owner PRESIDENT & FELLOWS OF HARVARD COLLEGE
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap