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Research Project:
Characterization, Production, and Utilization of Phytochemicals from
Agricultural Products
Location:
New Crops and Processing Technology Research
Title:
Separation of Soya Saponins Reference Standards by Kromaton Fast Centrifugal
Partition Chromatography
Authors
Glinski, Jan - PLANTA ANALYTICA, LLC
Davey, Mathew - PLANTA ANALYTICA, LLC
Berhow, Mark
Submitted to: International Soy Conference
Publication Acceptance Date: October 8, 2004
Publication Date: October 8, 2004
Citation: Glinski, J.A., Davey, M.H., Berhow, M.A. 2004. Separation
Of Soyba Saponins Reference Standards By Kromaton Fast Centrifugal Partition
Chromatography. International Soy Conference.
Technical Abstract: Centrifugal
Partition Chromatography (CPC) is an effective yet underutilized tool for
purification of compounds naturally produced by plants and microorganisms.
CPC separates compounds through partitioning between two immiscible liquid
phases of a bi-phasic solvent system. This technique does not involve
traditional solid chromatographic materials, known to affect adversely the
purification and yields of labile compounds. We have applied Kromaton FCPC (Kromaton
Technologies, France) to facilitate the purification of soya sapogenols (A,
B, and E) as well as saponins, of the type A and B. A one step fractionation
of a mixture of type B saponins in a specifically designed solvent system
led to the purification of some saponins and to obtaining highly enriched
fractions (50% ' 80%) of other saponins. Additional purification of the
enriched fractions requires usually another purification step. The method
provides an easy access to rare soya saponins, normally occurring only at
very low concentration. The FCPC purification is very well suited for
isolating of multi-gram quantities often needed for pharmacological or
clinical study.
Synthesis of Congeners and Prodrugs. 3. Water-Soluble Prodrugs of Taxol with
Potent Antitumor Activity
H. M. Deutsch, J. A. Glinski, M. Hernandez, R. d. Haugwitz,
School of Chemistry, Georgia Institute of Technology, Atlanta, Georgia
30332. Received October 19, 1987
Comparative Purification of Recombinant HIV-1 and HIV2 Reverse
Transcriptase: Preparation of Heterodimeric Enzyme Devoid of Unprocessed
Gene Product
Thomas C. Warren,*,l John J. Miglietta, Anthony Shrutkowski Janice M.
Roset Sher L. Rogers,* Klaus Lubbe,
Cheng
K. Shih,
t
Gary
O. Caviness, Richard lngraham, Deborah E. H. Palladino, Eva David,*
Grace C. Chow,11 Elizabeth B. Kopp,
t
Kenneth A. Cohen,:!: Jan A. Glinski,:!: Peter R. Farina,* and Peter M. Grob*
Departments of *Biochemistry, Molecular Biology, :!:Analytical Sciences,
§Medicinal Chemistry, and Immunology, Boehringer Ingelheim
Pharmaceuticals, Inc., Ridgefield, Connecticut 06877
Received February 3, 1992, and in revised form
August 3, 1992
Abstract
A procedure for producing and purifying recombinant HIV-l and HIV-2 reverse
transcriptase (RT) is described. These enzymes are produced by Escherichia
coli-transformed with a plasmid containing the gene en- coding for either
the human immunodeficiency virus type 1 (HIV-l) or HIV-2 RT protein. Both
proteins are partially processed by host cell proteases giving rise to a
mixture of heterodimeric and nonheterodimeric products, which are
subsequently resolved to near homogeneity by chromatography on
phosphocellulose, Q-Sepha rose, and hydrophobic interaction HPLC. Both HIV-l
(66/51 kDa) and HIV-2 (68/54 kDa) heterodimeric en zymes devoid of excess
unprocessed (p66 or p68) pre- cursors are isolated, enabling comparative
enzymatic characterization of the fully active (and biologically relevant)
heterodimeric forms. Homogenous HIV-l and HIV-2 RT purified by this
methodology exhibit near equivalent polymerase and RNase H activities. @
1992
Academic Press, Inc.
Screening natural products. Bioassay-directed isolation
of active components by centrifugal partition chromatography.
JAN A. GLINSKI, GARY O. CAVINESS, AND JULIE R.
MIKELL
Boehringer Ingelheim Pharmaceuticals. Inc. 175 Briar Ridge Road
Ridge/ield. Connecticut 06877
ABSTRACT
A Centrifugal Partition Chromatograph (CPC), Model LLN by Sanki, was used on
routine basis as a primary tool for over 200 assay-directed fractionations
in search of active principles from extracts of natural products. Various
extracts were found active in several receptor and enzymatic assays
incorporated into a high capacity screening system targeting discovery of
new anti-inflammatory, immunomodulating and antiviral agents. Centrifugal
partition chromatography is well suited for performing the assay-directed
fractionations, since like other countercurrent techniques it does not
involve solid phase adsorbents and thus is inherently less destructive. The
instrument was found reliable, simple to operate and applicable to the
entire range of polarity of natural products. It can handle milligram as
well as multigram quantities.
This method alone was applied to a mixture of saponins from an extract of
Alysicarpus sp. Isolation and subsequent identification of four new
saponins, Alysicarpins A, B, C, and D allowed a necessary in vitro
evaluation of their therapeutic potential. An extract of Psycho tria
acwninata, a tropical forest plant, gave inhibitory response in another
assay. Two crucial fractionation steps carried out by the CPC increased the
concentration of the active components.
Copyright
@ 1990 by Marcel Dekker,
Inc.
Reprinted from Journal of
Medicinal Chemistry, 1992,35. 1: Copyright @ 1992 by the American Chemical
Society and reprinted by permission of the copyright owner.
Effect of the 7-Amino Substituent on the Inhibitory Potency of
Mechanism-Based Isocoumarin Inhibitors for Porcine Pancreatic and Human
Neutrophil Elastases
A 1.85-A X-ray Structure of the Complex between Porcine Pancreatic Elastase
an( 7
-[ (N - Tosy
Ipheny lalany I )amino ]-4-chloro-3- methoxyisocoumarin
Maria A. Hernandez,t James C. Powers,..t
Jan Glinski,t Jozef Oleksyszyn,t J. Vijayalakshmi,t and Edgar F. Meyer, Jr.t
<
School of Chemistry and Biochemistry,
Georgia Institute of Technology, Atlanta, Georgia 30332-0400, and Department
of Biochemistry and Biophysics, Texas A
& M University, College Station, Texas
77843-2128. Received June 17, 1991
A series of new acyl, urea, and carbonate
derivatives of 7-amino-4-chloro-3-methoxyisocoumarin were synthesized and
evaluated as irreversible inhibitors of human neutrophil elastase (HNE) and
porcine pancreatic elastase (PPE). Inhibition of HNE is directly related to
the hydrophobicity of the substituent on the 7 -amino group. The N -
Tos- Phe derivative (19) is the best HNE inhibitor with a second-order rate
constant kobo/[I] = 200000 M-l S-l. The closest analogue in this
series, the 3,3-diphenylpropionyl derivative 5, had a kobo/[I] =
130000 M-l S-l with HNE. In contrast to the Tos-Phe derivative 19,
phenylacetyl derivative 2 and carbonates 22 and 25 gave extremely stable
enzyme-inhibitor complexes with deacylation half-lives longer than 48 h with
both elastases. N-Phenylurea derivative 25 was the best inhibitor for PPE
with a second-order rate constant kobo/[I] = 7300 M-l S-l. The
crystal structure of a complex of PPE with N-tosyl-Phe derivative 19 was
determined at 1.85-A resolution and refined to a final R factor of
16.9%. The isocoumarin forms an acyl enzyme with Ser-195, while His-57 is
near the inhibitor, but not covalently linked. The Tos-Phe makes a few
hydrophobic contacts with the S' subsites of PPE, but appears to be
interacting primarily with itself in the PPE structure. This region of HNE
is more hydrophobic and modeling indicates that the inhibitor would probably
make additional contacts with the enzyme.
Synthesis of
Congeners and Prodrugs.
3.1 Water-Soluble
Prodrugs of Taxol with Potent Antitumor Activity
H. M. Deutsch,t J. A. Glinski, M.
Hernandez, R. D. Haugwitz,~ v. L. Narayanan,~ M. Suffness,~ and L. H. Zalkow* School
of Chemistry, Georgia Institute of
Technology, Atlanta, Georgia 30332. Received October
19, 1987
Taxol has shown good in
vivo antitumor activity in a number of test systems. The formulation of
taxol for antitumor testing has been difficult. Esterification at either
C-2' or C- 7 resulted in loss of in vitro tubulin assembly activity but
not cytotoxicity. These observations suggested that esters at C-2' and/or
C-7, which would tend to promote water solubility, might serve as useful
prodrugs of taxol. The reaction of taxol with either succinic anhydride or
glutaric anhydride in pyridine solution at room temperature gave the
crystalline mono 2'-adducts 1b and If, respectively. Salts of these
acids (lb, If, Ii) were formed by the addition of 1 equiv of the
corresponding base, followed by evaporation and/ or freeze-drying of the
solvent(s). The salts had improved antitumor activity as compared to the
free acids. The triethanolamine and N-methylglucamine salts showed greatly
improved aqueous solubility and were more active than the sodium salts. The
glutarate series was preferred because of the higher activity and the higher
yields obtained. 2'-Glutaryltaxol (If) was coupled with
3-(dimethylamino)-1-propylamine, using Cill, to form in excellent yield the
amino amide 10. The hydrochloride salt (lp) showed good solubility and was
extremely potent and active. At 10 mg/kg, in the B16 screen, 1p gave
a T /C of 352 with 5 out of 10 cures. In the MX-1 breast xenograft assay,
this prodrug gave values of -100 at doses of 40 and 20 mg/kg, with all live
animals being tumor free.
The Structure and Partial Synthesis of Delalatine, an Alkaloid from Delphinium
Species.
SAMIR A. Ross, HARIDUTT
K. DESAI, BALAWANT S. JOSHI, SANTOSH K. SRIVASTAVA, JAN A. GLINSKI, SI YING
CHEN and S. WILLIAM PELLETIER *
Institute for Natural
Products Research and the School of Chemical Sciences, The University of
Georgia, Athens, Georgia 30602, U.S.A.
(Received
1 March 1988)
Key Word
Index-Delphinium elatum; D. tatsienense; Ranunculaceae;
delelatine; C19-diterpenoid alkaloid.
Abstract-A new
C-19-diterpenoid alkaloid designated as delelatine has been isolated from
Delphinium elatum L. and D. tatsienense Franch. and its structure
has been elucidated by spectroanalytical methods. Delelatine has been
correlated with 14-acetyl-l0-deoxydictyocarpine and synthesized from
dictyocarpine, thus confirming its structure and stereochemistry.
Centrifugal
Partition Chromatography in Assay-Guided Isolation of Natural Products: A
Case Study of Immunosuppressive Components of Tripterygium wilfordii
This outstanding guide introduces centrifugal partition
chromatography (CPC) for any biphasic system—offering in-depth coverage of
instrumentation, theory, liquid-liquid partition coefficients, and CPC in
organic and inorganic chemistry.
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