US20060229279A1 - Artemisinins with improved stability and bioavailability for therapeutic drug development and application - Google Patents

Artemisinins with improved stability and bioavailability for therapeutic drug development and application Download PDF

Info

Publication number
US20060229279A1
US20060229279A1 US11450009 US45000906A US2006229279A1 US 20060229279 A1 US20060229279 A1 US 20060229279A1 US 11450009 US11450009 US 11450009 US 45000906 A US45000906 A US 45000906A US 2006229279 A1 US2006229279 A1 US 2006229279A1
Authority
US
Grant status
Application
Patent type
Prior art keywords
cyclodextrin
acid
artelinic
β
fig
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Abandoned
Application number
US11450009
Inventor
Mark Hartell
Apurba Bhattacharjee
Rickey Hicks
John VanHamont
Wilbur Milhous
Original Assignee
Hartell Mark G
Bhattacharjee Apurba K
Hicks Rickey P
Vanhamont John E
Milhous Wilbur K
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date

Links

Images

Classifications

    • BPERFORMING OPERATIONS; TRANSPORTING
    • B82NANOTECHNOLOGY
    • B82YSPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
    • B82Y5/00Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
    • A61K31/00Medicinal preparations containing organic active ingredients
    • A61K31/33Heterocyclic compounds
    • A61K31/395Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine, rifamycins
    • A61K31/55Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine, rifamycins having seven-membered rings, e.g. azelastine, pentylenetetrazole
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
    • A61K31/00Medicinal preparations containing organic active ingredients
    • A61K31/70Carbohydrates; Sugars; Derivatives thereof
    • A61K31/715Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
    • A61K31/716Glucans
    • A61K31/724Cyclodextrins
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
    • A61K47/00Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/69Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit
    • A61K47/6949Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit inclusion complexes, e.g. clathrates, cavitates or fullerenes
    • A61K47/6951Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit inclusion complexes, e.g. clathrates, cavitates or fullerenes using cyclodextrin
    • YGENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02ATECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection
    • Y02A50/30Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
    • Y02A50/38Medical treatment of vector-borne diseases characterised by the agent
    • Y02A50/408Medical treatment of vector-borne diseases characterised by the agent the vector-borne disease being caused by a protozoa
    • Y02A50/411Medical treatment of vector-borne diseases characterised by the agent the vector-borne disease being caused by a protozoa of the genus Plasmodium, i.e. Malaria
    • YGENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10STECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10S514/00Drug, bio-affecting and body treating compositions
    • Y10S514/895Malaria

Abstract

A stable form of artemisinin wherein an artelinic acid or artesunic acid is complexed with cyclodextrin analogs, preferably, β-cyclodextrin. The complexed cyclodextrin artemisinin formulation shields the peroxide portion of the artemisinin backbone from hydrolytic decomposition rendering it stable in solution. Artelinic acid and cyclodextrin are placed into contact with one another to yield a 2:1 molecular species. Artesunic acid and cyclodextrin yield a 1:1 molecular species. The complexed cyclodextrin artemisinin formulation is effective for the treatment of malaria and is stable in solution for long periods of time.

Description

  • [0001]
    This application claims priority of provisional application No. 60/362,985 filed Mar. 7, 2002.
  • GOVERNMENT INTEREST
  • [0002]
    The invention described herein may be manufactured, used and licensed by or for the U.S. Government.
  • BACKGROUND OF THE INVENTION
  • [0003]
    1. Field Of The Invention
  • [0004]
    A novel form of artemisinins that are complexed with cyclodextrin for solving stability problems associated with previous forms of artemisinins.
  • [0005]
    2. Brief Description Of Related Art
  • [0006]
    Artelinic acid is an effective antimalarial agent when in contact with the malarial parasite. However, artelinic acid has poor stability in solution and, thus, has limited bioavailability in vivo. Artemisinins, as a class, include such analogs as artelinic acid and artesunic acid among many others. Currently, no analog of the artemisinin class of compounds exists which can remain stable in solution. Injectable formulations of artemisinin analogs, such as artelinic acid and artesunic acid, are not FDA approved due to their instability in solution. All artemisinins contain a peroxide bridge susceptible to hydrolytic cleavage. Artemisinins have been found to yield an inferior class of antimalarials due to these severe limitations in chemical stability. Artemisinins are limited to only being packaged as solids for oral dosing, as previous patents have claimed. U.S. Pat. Nos. 6,326,023; 6,307,068; 6,306,896; 5,834,491; 5,677,331; 5,637,594; 5,486,535; 5,278,173; 5,270,037; 5,219,865; 5,021,426; 5,011,951.
  • [0007]
    Application of an antimalarial formulation must be specific to administration in hot, humid tropical regions native to the malarial parasite. Thus, chemical stability under drastic environmental conditions is essential. Attempts to produce a more stable form of artelinic acid have been accompanied by critical limitations. A soluble sodium salt of artelinic acid has been successfully formulated, but eventually degrades over time. This is presumably due to a re-formation of the insoluble acid. Numerous attempts at preventing this precipitate have been unsuccessful.
  • [0008]
    The osmolality of the salt solution is significantly less than the predicted value indicating possible inter-molecular complexation that may be responsible for eventual precipitation over time. An amine-based buffer of artelinic acid has been successfully formulated, but yields a higher pH solution (>8.0) that induces significant vein irritation upon injection. Additional localized redness and swelling surrounding the injection site is a notable contraindication to a preferred intravenous formulation. Additionally, amine-based buffers have been observed to take on a strong yellow hue over time. The mechanism of color formation has not been deduced, but implies a modification of the artelinate formulation, which is not conducive to pharmaceutical preparations where a defined constant state of purity is essential.
  • [0009]
    U.S. Pat. Nos. 6,326,023; 6,307,068; 6,306,896; 5,834,491; 5,677,331; 5,637,594; 5,486,535; 5,278,173; 5,270,037; 5,219,865; 5,021,426; 5,011,951 are only directed to be packaged as solids for oral dosing.
  • [0010]
    Therefore, there is a need to provide a form of artemisinins that solve the stability problems associated with previous formulations.
  • [0011]
    It is an object of the present invention to provide a form of artemisinins, such as but not limited to artelinic acid and artesunic acid that solves the stability problems associated with previous formulations.
  • [0012]
    It is another object of the present invention to provide a stable form of artemisinins that is injectable.
  • [0013]
    It is still another object of the present invention to provide a stable form of artemisinins that does not develop a yellow hue over time.
  • [0014]
    It is still another object of the invention to promote bioavailability and membrane permeability while decreasing the likelihood of localized inflammation at the route of entry, thus increasing its therapeutic activity.
  • [0015]
    These and other objects of the invention will become apparent upon a reading of the entire disclosure.
  • SUMMARY OF THE INVENTION
  • [0016]
    The present invention is directed to cyclodextrin complexed with artelinic acid or artesunic acid to form complexed cyclodextrin-artemisinin formulations in a 2:1 ratio of cyclodextrin per artelinic acid molecule or in a 1:1 ratio of cyclodextrin per artesunic acid molecule. The formulation is stable in solution, bioavailable, membrane permeable and does not cause inflammation upon injection.
  • BRIEF DESCRIPTION OF THE FIGURES
  • [0017]
    FIG. 1 is a plot of the hypsochromic shift observed with increasing concentrations of cyclodextrin. Artelinic acid concentration=10 mM.
  • [0018]
    FIG. 2 a is an absorption spectrum of 10 mM artelinic acid with and without 1 mM β-cyclodextrin;
  • [0019]
    FIG. 2 b is an absorption spectrum of 10 mM artelinic acid with and without 4 mM β-cyclodextrin;
  • [0020]
    FIG. 3 is a 600 MHz WATERGATE-TOCSY NMR spectrum of 1.2 mM artelinic acid with 2.5 mM β-cyclodextrin in PBS (pH 7.4);
  • [0021]
    FIG. 4 is a 600 MHz WATERGATE-ROESY NMR spectrum of 1.2 mM artelinic acid with 2.5 mM β-cyclodextrin in PBS (pH 7.4);
  • [0022]
    FIG. 5 is a 600 MHz WATERGATE-ROESY NMR spectrum of 1.2 mM artelinic acid with 2.5 mM β-cyclodextrin in PBS (pH 7.4);
  • [0023]
    FIG. 6 is a 600 MHz WATERGATE-ROESY NMR spectrum of artesunate with an excess of β-cyclodextrin in PBS (pH 7.4);
  • [0024]
    FIG. 7 a is the aromatic region of the 600 MHz proton spectra of 1.2 mM artelinic acid;
  • [0025]
    FIG. 7 b is the aromatic region of the 600 MHz proton spectra of 1.2 mM artelinic acid complexed with 2.5 mM β-cyclodextrin in PBS (pH 7.4);
  • [0026]
    FIG. 8 a is the alkyl region of the 600 MHz proton NMR spectra of 1.2 mM artelinic acid;
  • [0027]
    FIG. 8 b is a 600 mHz proton NMR spectrum of 1.2 mM artelinic acid complexed with 2.5 mM β-cyclodextrin in PBS (pH 7.4);
  • [0028]
    FIG. 9 a is a 600 MHz proton NMR spectrum of 2.5 mM β-cyclodextrin in PBS (pH 7.4);
  • [0029]
    FIG. 9 b is a 600 MHz proton NMR spectrum of 2.5 mM β-cyclodextrin with 1.2 mM artelinic acid in PBS (pH 7.4);
  • [0030]
    FIG. 10 a is a 600 MHz proton NMR spectrum (protons number 2 to 6) of 2.5 mM β-cyclodextrin in PBS (pH 7.4);
  • [0031]
    FIG. 10 b is a 600 MHz proton NMR spectrum (protons number 2 to 6) of 2.5 mM β-cyclodextrin complexed with 1.2 mM artelinic acid in PBS (pH 7.4);
  • [0032]
    FIG. 10 c is a 600 MHz proton NMR spectrum (protons number 2 to 6) of artesunate with an excess of β-cyclodextrin in PBS (pH 7.4);
  • [0033]
    FIG. 11 is a 600 MHz proton NMR spectrum of 2.5 mM β-cyclodextrin and 1.2 mM artelinic acid in PBS buffer at pH 7.4 with 1:9 D2O/H2O;
  • [0034]
    FIG. 12 is a 2D NOESY spectrum of 2.5 mM β-cyclodextrin and 1.2 mM artelinic acid in PBS buffer at pH 7.4 with 1:9 D2O/H2O;
  • [0035]
    FIG. 13 is a 600 MHz proton NMR spectrum of artelinic acid BN BPI 1387, WR#255663;
  • [0036]
    FIG. 14 is a 600 MHz proton NMR spectrum of 2D TOESY spectrum of 2.5 mM β-cyclodextrin and 1.2 mM artelinic acid in PBS buffer at pH 7.4 with 1:9 D20/H2O;
  • [0037]
    FIG. 15 is a 600 MHz proton NMR spectrum of 2D ROESY spectrum of 2.5 mM β-cyclodextrin and 1.2 mM artelinic acid in PBS buffer at pH 7.4 with 1:9 D2O/H2O;
  • [0038]
    FIG. 16 is a 600 MHz proton NMR spectrum of artesunate with an excess of β-cyclodextrin in PBS buffer at pH 7.4;
  • [0039]
    FIG. 17 is a 600 MHz proton NMR spectrum of 2D ROESY spectrum of artesunate with an excess of β-cyclodextrin in PBS buffer at pH 7.4;
  • [0040]
    FIG. 18 a is the electrostatic potential map of the primary face of β-cyclodextrin looking into the molecule from the top;
  • [0041]
    FIG. 18 b is the electrostatic potential map of the primary face of β-cyclodextrin as shown in FIG. 18 a rotated to the left;
  • [0042]
    FIG. 18 c is the electrostatic potential map of the secondary face of β-cyclodextrin;
  • [0043]
    FIG. 18 d is a molecular model of FIG. 18 d illustrating the positions of specific atoms;
  • [0044]
    FIG. 19 a is a side view of the electrostatic potential map of artelinic acid;
  • [0045]
    FIG. 19 b is a rear view of the electrostatic potential map of artelinic acid;
  • [0046]
    FIG. 20 is the electrostatic potential map of β-cyclodextrin complexed with artelinic acid in a 2:1 molecular ratio;
  • [0047]
    FIG. 21 is a molecular model of β-cyclodextrin complexed with artelinic acid in a 2:1 molecular ratio showing degrees of insertion and interaction between each molecule;
  • [0048]
    FIG. 22 is an axial view from the primary face of the electrostatic potential map of β-cyclodextrin complexed with artelinic acid in a 2:1 molecular ratio indicating the electrostatic interaction between the benzoic acid moiety and one of the cyclodextrins;
  • [0049]
    FIG. 23 is a plot of osmolality versus concentration of artelinate in aqueous solution compared to theoretical determinations based on the complete disassociation of the salt;
  • [0050]
    FIG. 24 is a plot of osmolality versus concentration of a lysine-artelinate salt preparation in aqueous solution compared to theoretical determinations based on the complete disassociation of the salt;
  • [0051]
    FIG. 25 is a plot of osmolality versus concentration of a lysine-artelinate salt preparation with 3 molar equivalents of lysine in aqueous solution compared to theoretical determinations based on complete disassociation of the salt;
  • [0052]
    FIG. 26 is the linear regression (R=0.994, p<0.0001) of experimentally measured osmolality of artelinate complexed with hydroxypropyl-β-cyclodextrin (1:2 mole ratio) in aqueous solution. Upper and lower 95% confidence intervals and 95% prediction limits are also indicated;
  • [0053]
    FIG. 27 a-c are plots of relative deviation between experimentally measured osmolality and theoretical determinations based on complete disassociation for 3 aqueous artelinate formulations: lysine-artelinate prepared with 1 molar equivalent of lysine, lysine-artelinate prepared with 3 molar equivalents of lysine, and hydroxypropyl-β-cyclodextrin-artelinate (2:1) complex;
  • DETAILED DESCRIPTION
  • [0054]
    The present invention is directed to a novel form of artemisinins that remain stable over time in solution. The artemisinins may be, but are not limited to artelinic acid and artesunic acid. This novel form of artemisinins uses a unique complexed form of the therapeutic agent with cyclodextrin analogs, such as but not limited to alpha-, beta-, and gamma-cyclodextrin analogs and their derivatives.
  • [0055]
    The present invention is directed to cyclodextrin complexed with artelinic acid in a 2:1 ratio which is a form of artemisinin that alters the electron cloud surrounding the artemisinin molecule in such a way as to stabilize this agent to promote bioavailability and membrane permeability while decreasing the likelihood of localized inflammation at the route of entry. Thus, this form of artemisinin increases its therapeutic activity. Artesunic acid was complexed with cyclodextrin, but in a unique 1:1 ratio in such a way as to stabilize the agent yield similar increases in its therapeutic activity.
  • [0056]
    The stability of the artemisinins is achieved by changing the physiocochemical properties such as but not limited to electron density, electrostatic potential and charge transfer mediated complexation.
  • [0057]
    The complexed cyclodextrin formulation of the artemisinins described deliberately shields the peroxide bridge of the artemisinin backbone from hydrolytic decomposition. Additionally, the aromatic benzoic acid portion of the artelinate molecule is also complexed with a second cyclodextrin molecule. This unique 2:1 complexation with cyclodextrin is not intuitively obvious because artelinic acid alone is unstable in aqueous solution. Simply placing cyclodextrin in solution with artelinic acid would not achieve these results, as the artelinic acid would not be in contact with the cyclodextrin to form complexation. Further, cyclodextrin is know to form complexes with itself and thus may not be readily available in solution to interact efficiently and effectively with the artelinic acid. The inventors have placed artelinic acid and cyclodextrin into contact with one another and have complexed them in such a manner as to yield a stable 2:1 molecular species. The inventors have also placed artesunic acid and cyclodextrin into contact with one another and have complexed them in such a manner as to yield a stable 1:1 molecular species.
  • [0058]
    The present molecules are stable under ambient or physiologically relevant conditions.
  • [0000]
    Materials And Methods
  • [0059]
    β-cyclodextrin was obtained from Sigma-Aldrich Corp., St. Louis, Mo. Artelinic acid was alkalinized with NaOH to yield the sodium salt. Standardized PBS buffer at a pH of 7.4 was obtained from Invitrogen Corp., Carlsbad, Calif.
  • [0060]
    Absorption Spectroscopy Studies.
  • [0061]
    Mixtures of artelinate (10 μM) were prepared with increasing concentrations of β-cyclodextrin (0.0, 1.0, 4.0, 6.0, and 9.0 mM). Absorption spectra were collected on a Beckman DU Series 600 Spectrophotometer.
  • [0062]
    The spectra collected indicated a clear hypsochromic or blue shift in the absorption maximum at 230 nm with increasing concentrations of cyclodextrin. Hypochromic effects were also notable at 230 nm, as well as the broader transitions observed at 275 and 382 nm (FIG. 1). This combined observation is consistent with inclusion interactions of the benzoic anion of artelinate with cyclodextrin.
  • [0063]
    Changes in observed isosbestic points at higher cyclodextrin concentrations indicates a complicated molecular species containing greater than a simple 1:1 molecular species (FIGS. 2 a and 2 b).
  • [0064]
    1H NMR Studies.
  • [0065]
    Mixtures of β-cyclodextrin (2.5 mM) and artelinic acid (1.2 mM) were prepared in PBS (pH 7.4) and incubated at 37° C. for 2-3 hour to promote complexation prior to analysis.
  • [0066]
    All 1H NMR data was collected using a Bruker DRX-600 spectrometer operating at a proton frequency of 600.02 MHz at a temperature of 25° C. Solvent suppression was accomplished by application of the WATERGATE (WATER suppression by GrAdient Tailored Excitation) pulse sequence developed by Sklenar and co-workers. This sequence provides excellent suppression of the water resonance by a combination of rf pulses and a series of gradient pulses. The sequence combines a non-selective 90° pulse with a symmetrical echo formed by two short gradient pulses in conjunction with a 180 selective (on water) pulse train.
  • [0067]
    The two-dimensional WATERGATE-TOCSY experiment employed a modified MLEV-17 spin-lock sequence for a total mixing time of 80 ms, including the 2.5 ms trim pulses at the beginning and the end of the spin-lock. The spectrum was collected with a spectral width of 7183.91 Hz (11.972 ppm) using 2K data points with 32 scans per 256 t1 increments with a 1.5 s recycle delay. The data was processed by multiplication with a 90° shifted sine-bell window function in each dimension, with one zero fill in the f1 dimension before transformation to produce matrices consisting of 512 data points in both dimensions.
  • [0068]
    The two-dimensional WATERGATE-NOESY spectra were collected with a spectral width of 7183.91 Hz (11.972 ppm) using 2K data points with 128 scans per 512 t1 increments with a 1.5 s recycle delay. The data was processed by multiplication with a 90° shifted sine-bell window function in each dimension, with one zero fill in the f1 dimension before transformation to produce matrices consisting of 512 data points in both dimensions. Two different experiments were conducted with mixing times of 50 and 600 ms.
  • [0069]
    The two-dimensional WATERGATE-ROESY spectrum was collected with a spectral width of 7183.91 Hz (11.972 ppm) using 2K data points with 256 scans per 512 t1 increments with a 1.5 s recycle delay with a spin-lock mixing pulse of 400 ms. The data was processed by multiplication with a 90° shifted sine-bell window function in each dimension, with one zero fill in the f1 dimension before transformation to produce matrices consisting of 512 data points in both dimensions.
  • [0070]
    Two-dimensional NMR methods were used to determine the degree of capping or complexation of artelinic acid by β-cyclodextrin. The 2D WATERGATE-TOSCY spectrum of artelinic acid (FIG. 3) clearly indicates that the individual spin-spin coupling networks of a mixture of artelinic acid and β-cyclodextrin can be resolved. In FIG. 3, the spin-spin coupling network for β-cyclodextrin is shown at A and the spin-spin coupling network for the alkyl ring of artenilate is shown at B. The 2D-rotating frame NOE spectrum, WATERGATE-ROESY, of artelinic acid was collected at a mixing time of 400 ms and is shown in FIG. 4. The labeled intermolecular ROE interaction between the aromatic protons of artelinic acid with both the anomeric and ring protons of β-cyclodextrin proves that this region of artelinic acid is complexed with one molecule of β-cyclodextrin. In FIGS. 4, A, B and C indicate the intermolecular dipolar ROE coupling between the aromatic protons of artelinate with the glucose ring protons of β-cyclodextrin. The ROE between the meta protons are more intense than those observed for the ortho protons indicating that meta protons are inserted deeper into the cavity. D and F indicate the dipolar coupling between the ortho protons of artelinate with the two benzyl protons of artelinate. E indicates the dipolar coupling between the meta protons of artelinate with the anomeric protons of β-cyclodextrin. FIG. 5 shows the alkyl region of this same spectrum. The labeled intermolecular ROE's between the alkyl ring protons of artelinic acid with both the anomeric and ring protons of β-cyclodextrin indicate that this region of artelinic acid is complexed with one molecule of β-cyclodextrin. These observations are similar to those reported by Nishijo (Nishijo, J.; Nagai, M.; Yasuda, M.; Ohno, E.; Ushiroda, Y. J. Pharm. Sci. 1995, 84, 1420-1426) and by Redenti (Redenti, E.; Ventura, P.; Fronza, G.;Selva, A.;Rivara, S.;Plazzi, P. V.; Mor, M. J. Pharm. Sci. 1999, 88, 599-607) in similar NMR β-cyclodextrin complexation studies. In FIG. 5, A represents a region that contains the dipolar coupling between the ring protons of β-cyclodextrin and the alkyl ring proton of artelinate; and B represents the region that contains the dipolar coupling of the anomeric protons of β-cyclodextrin with the alkyl protons of artelinate.
  • [0071]
    Two 2D WATERGATE-NOESY spectra were collected at mixing times of 50 and 600 ms (data not shown). The NOESY spectrum collected at 600 ms gave similar intermolecular and intramolecular NOE's to those observed in the ROESY spectrum, however the observed intensities were reduced. The NOESY spectrum collected at 50 ms did not exhibit the intermolecular NOE's between artenilate and β-cyclodextrin. This observation is consistent with what one would expect due to the fact that intermolecular NOE's require a longer mixing time to develop as compared to intramolecular NOE's.
  • [0072]
    The 2D ROESY and NOESY data clearly indicate that both the alkyl and aromatic regions of artelinic acid are complexed with one individual molecule of β-cyclodextrin.
  • [0073]
    In FIG. 6, the spectrum of artesunate with an excess of β-cyclodextrin in PBS is shown. This data clearly indicates that the artesunate is capped by β-cyclodextrin in a 1:1 ratio. The region that is represented by A contains the intramolecular dipolar coupling the alkyl ring proton of artesunate. The region that is represented by B contains the intermolecular dipolar coupling the alkyl ring proton of artesunate with the ring protons of β-cyclodextrin. The region that is represented by C contains the intermolecular dipolar coupling the alkyl ring proton of artesunate with the anomeric protons of β-cyclodextrin. The region that is represented by D contains additional intramolecular dipolar coupling the alkyl ring proton of artesunate. The region that is represented by E contains the intramolecular dipolar coupling of the β-cyclodextrin.
  • [0074]
    FIG. 7 a shows the aromatic region of the 600 MHz proton spectra of 1.2 mM artelinic acid and FIG. 7 b is the aromatic region of the 600 MHz proton spectra of 1.2 mM artelinic acid complexed with 2.5 mM β-cyclodextrin. Upon complexation the aromatic resonances of artelinate are both shifted upfield. The chemical shift values and the relative changes in chemical shift values are given in Table 1. A similar shift of aromatic protons resonances of ketoconazole on complexation with β-cyclodextrin was reported by Redenti and co-workers (Redenti, E.; Ventura, P.; Fronza, G.;Selva, A.;Rivara, S.;Plazzi, P. V.; Mor, M. J. Pharm. Sci. 1999, 88, 599-607). In addition, the intensity of the resonance for protons 2 and 2′ is reduced indicating complexation.
    TABLE 1
    1H Chemical Shift Assignments (δ) for the Aromatic Protons
    and Methyl Protons of Artelinic Acid
    Chemical Shift
    complexed with
    Proton Chemical Shift β-cyclodextrin Δδ (ppm)
    3 and 3′ 8.09 7.82 +0.27
    2 and 2′ 7.42 7.25 +0.17
    methyl #1 0.98 1.02 −0.04
    methyl #2 0.95 0.95 0.00
  • [0075]
    FIG. 8 a shows the alkyl region of the 600 MHz proton spectra of 1.2 mM artelinic acid and FIG. 8 b shows 1.2 mM artelinic acid complexed with 2.5 mM β-cyclodextrin. As seen from these spectra the chemical shift position and the appearance of the methyl protons have changed indicating complexation of this region of the molecule with β-cyclodextrin. The chemical shift of the resonances for methyl group #1 are shifted upfield by 0.04 ppm (Table 1). The resonances for both methyl groups were broadened and less well resolved.
  • [0076]
    FIG. 9 a is a 600 MHz proton spectra of 2.5 mM β-cyclodextrin and FIG. 9 b is a 600 MHz proton spectra of 2.5 mM β-cyclodextrin with 1.2 mM artelinic acid. These spectra clearly indicate that chemical values for protons 2 to 6 on β-cyclodextrin change on complexation with artelinic acid. Similar shifts in the proton resonances for β-cyclodextrin have been reported by Nishijo and co-workers (Nishijo, J.; Nagai, M.; Yasuda, M.; Ohno, E.; Ushiroda, Y. J. Pharm. Sci. 1995, 84, 1420-1426).
  • [0077]
    FIG. 10 a-10 c show the proton spectra (protons number 2 to 6) of 2.5 mM β-cyclodextrin, 2.5 mM β-cyclodextrin complexed with 1.2 mM artelinic acid and 1.2 mM artesunate in an excess of β-cyclodextrin, respectively. These spectra clearly indicate a different mode of complexation for the two artemisinin analogs.
  • [0078]
    Table 2 summarizes the chemical shift assignments for cyclodextrin compared with the corresponding complexes with artelinic acid and artesunic acid as derived from FIGS. 9 and 10. The change in chemical shifts (Δδ) clearly demonstrate that both cyclodextrins of the artelinic acid complex and the cyclodextrin of the artesunic acid complex coordinate at the 3-H end or secondary face (FIG. 18) of the cyclodextrin. Further, the benzoic acid moiety of artelinic acid coordinates deeply into the cyclodextrin pocket yielding significant changes in chemical shift for the 3-H, 5-H, and 6-H protons. In contrast, artesunic acid, which only binds to one cyclodextrin at the peroxide bridge, produced chemical shift changes of a lower magnitude indicating a more shallow binding interaction. Lastly, for the artesunate-cyclodextrin complex the changes in chemical shift indicate Δδ of 6H<5H<3H which clearly demonstrates this shallow binding interaction compared to the deep insertion of the benzoic acid moiety of artelinic acid. This data clearly supports a unique stereochemical arrangement based upon the physicochemical properties of each molecular species to yield a specific stable complex.
    TABLE 2
    1H Chemical Shift Assignments (δ) for
    the Cyclodextrin Protons (2 through 6)
    2H 3H 4H 5H 6H
    β-cyclodextrin 3.63 3.94 3.56 3.83 3.86
    artelinic acid 3.61 3.83 3.53 3.72 3.74
    Δδ′ 0.02 0.11 0.03 0.11 0.12
    β-cyclodextrin 3.63 3.94 3.56 3.83 3.86
    artesuate 3.62 3.88 3.55 3.79 3.84
    Δδ 0.01 0.06 0.01 0.04 0.02
  • [0079]
    FIGS. 11 through 17 provide ancillary and supportive data that was used in elucidating the structural conformation of the described cyclodextrin complexes.
  • [0080]
    Molecular Electrostatic Potential Mapping and Docking/Affinity Determinations.
  • [0081]
    Molecular Electrostatic Potential (MEP) maps on cyclodextrin and artelinic acid were developed by calculating electrostatic potentials on the van der Waals surface of the molecules using the semi-empirical PM3 molecular orbital theory as implemented in the SPARTAN software (SPARTAN version 4.0, Wavefunction, Inc., 18401 Von Karman Ave., #370, Irvine, Calif. 92715 U.S.A. 1995 Wavefunction, Inc.). PM3 is a semi-empirical quantum chemical theory model based on Thiel's integral formalism underlying MNDO/d, and is used in conjunction with parameters for both transition and non-transition metals (reference: (a) W. Thiel and A. Voityuk, Theor. Chim. Acta., 81, 391, (1992); (b) W. Thiel and A. Voityuk, Int. J. Quantum Chem., 44, 807 (1992).
  • [0082]
    Molecular electrostatic potential (MEP) maps and their electrostatic potential energy isopotential profiles were generated and sampled over the entire accessible surface of a molecule (corresponding roughly to a van der Waals contact surface). The MEP maps provide a measure of charge distribution from the point of view of an approaching reagent. This is calculated using a test positive charge as the probe. Thus, these types of profiles can provide an estimate of electronic distribution surrounding the molecule so as to enable qualitative assessment of any possible interaction with an approaching molecule. However, conformation search calculations using the “systematic search” technique via the single-point PM3 method of SPARTAN were used to generate different conformers for each of the molecules. The minimum energy conformer with highest abundance (a Boltzman population density greater than 70.0%) was chosen for full geometry optimization using the PM3 algorithm. The MEP profiles were generated on the optimized geometry of the molecules. The computations were carried out on a Silicon Graphics Octane workstation.
  • [0083]
    To further understand the binding affinities between cyclodextrin and artelinic acid, the complete optimized structures of both the compounds have been considered and docking calculations using the Docking/affinity module in Insight II (Accelrys Inc., 9685 Scranton Road, San Diego, Calif. 92121-3752) were conducted. See Oprea, T. I. and Marshall, G. R. (1998) Receptor-based prediction of binding affinities. Perspectives in Drug Discovery and Design 9/10/11:35-61; and Insight II User Guide, San Diego: Accelrys Inc. (2002), which are herein incorporated by reference.
  • [0084]
    Docking/affinity module in Insight II allows calculating the nonbonded energy between two molecules using explicit van der Waals energy, explicit electrostatic (Coulombic) energy, or both van der Waals and electrostatic energies. The number of atoms included in the calculation can be limited by specifying a monomer- or residue-based cutoff. Other methods known in the art may be used, for example, the computation can be done using a pre-computed energy grid.
  • [0085]
    These molecular modeling determinations based on unique and specific physicochemical properties of the artemisinins studied complexed with β-cyclodextrin produced conceptual models which clearly rationalized the direct physical measurements of the NMR experiments. FIGS. 18 a-d illustrate the unique electrostatic potential map of β-cyclodextrin showing the primary binding faces (FIGS. 18 a and 18 b) and secondary binding faces (FIGS. 18 c and FIG. 18 d). Most notable is the unique net positive region 1 of the electron cloud at the primary face.
  • [0086]
    FIGS. 19 a and 19 b illustrate the unique electrostatic potential map of artelinic acid. Most notable is the dense negative region 2 of the carboxylic acid tail as well as a more subtle negative region 3 of the peroxide bridge.
  • [0087]
    FIG. 20 clearly demonstrates the 2:1 complexation of β-cyclodextrin with artelinic acid. Two β-cyclodextrin molecules are shown at 4 and one artelinic acid molecule is shown at 5. The depth of insertion of the carboxylic acid tail compared to the peroxide bridge portion of the molecule is more clearly illustrated in the corresponding ball-and-stick model of the complex in FIG. 2 lwherein two β-cyclodextrin molecules are shown at 4 and one artelinic acid molecule is shown at 5.
  • [0088]
    Lastly, FIG. 22 directly illustrates the unique physicochemical interaction of the electrostatc potential map of cyclodextrin with that of the artelinic acid tail. This axial view into the primary face of the second cyclodextrin molecule clearly illustrates this unique and selective electrostatic interaction. The negative region of the electrostatic potential map is shown at 6 and the positive region of electrostatic potential map is shown at 7.
  • [0089]
    Simple docking calculations do not yield these results as they assume an in vacuo environment. Inclusion complexes with cyclodextrins are mediated by the release of high-energy water molecules from the inner core of the cyclodextrin molecule. Therefore, direct structural measurements of the complex by techniques such as high resolution multi-dimensional NMR rationalized by physicochemical property determinations such as but not limited to molecular electrostratic potential mapping is specifically required to accurately characterize these complexes.
  • [0090]
    Osmometry Determinations.
  • [0091]
    Solutions of hydroxypropyl-β-cyclodextrin and artelinic acid of varied compositions as indicated were measured at room temperature using a Fiske ONE-TEN Osmometer (Fiske Associates, Norwood Mass., USA). The solvent for all experiments was ultra-pure distilled deionized water (18 MΩ) filtered through a 0.45 μm filter. Small sample volumes (15 μL) were measured in units of mOsmol/kg water with an instrument repeatability of ±2 mOsmol/kg water in the data range studied (0 to 400 mOsmol/kg water). The instrument was calibrated routinely with NIST standards of NaCl and a daily NIST reference of NaCl was verified at the start of each set of experiments.
  • [0092]
    Osmolality is a direct measure of the degree of molecular dissociation of a species in water. FIG. 23 illustrates the deviation of measured osmolality in aqueous artelinate solutions versus theoretical calculations which assume complete dissociation. This deviation from ideality also appears to have a significant margin of error as observed by the marked degree of data scatter in the measurments.
  • [0093]
    FIGS. 24 and 25 illustrate a similar relationship between measured osmolality and ideal dissociation with a lysine salt formulation and a lysine salt formulation with 3 molar equivalents excess lysine. All three artelinate formulations appear to deviate strongly from ideality. Secondly, the measure of osmolality versus concentration of artelinate appears to be biphasic as demonstrated most clearly in FIG. 25, but also observed in FIGS. 23 and 24.
  • [0094]
    FIG. 26 illustrates the strong linear correlation of the experimentally measured osmolality of artesunate complexed with hydroxypropyl-β-cyclodextrin in aqueous solutions. Hydroxypropyl-β-cyclodextrin was chosen for all osmolality determinations, as its aqueous solubility is greater than β-cyclodextrin and its well-established pharmacological compatibility for future i.v. drug formulations.
  • [0095]
    Measured deviation in osmolality of the artelinic acid-cyclodextrin ( 1:2) formulation after 28 days at room temperature was <7% in the concentration range of 15-25 mg/mL artelinate. This 7% deviation was consis tently observed as an increase in osmolality due to an enhancement of solvation over time, rather than a decrease in solubility. The more concentrated solutions of cyclodextrin complexes would need to incubate for longer periods of time to ensure maximum complexation.
  • [0096]
    FIGS. 27 a-c illustrate the deviations from ideality of three artelinate formulations, 1 molar equivalent of lysine shown at FIG. 27 a, lysine-artelinate prepared with 3 molar equivalens of lysine shown at FIG. 27 b and cyclodextrin-artelinate (2: 1) complex shown at FIG. 27 c. The artelinate-cyclodextrin formulation clearly deviates from ideality in a more predictable manner. The decrease in relative deviation with increasing concentration is mostly likely due to enhanced complexation due to a Le Chatelier's shift in solution equilibrium. This is notably contrasted with the other two formulations which yield solutions that deviate in an increasing manner (10-15%) from 12 to 30 mg/mL.
  • [0097]
    Injectable Formulation:
  • [0098]
    The stable form of artemisinin, the cyclodextrin complexed with artenilate in a 2:1 ratio, may be dissolved in saline, phosphate buffered saline (PBS), deionized water or any other suitable aqueous carrier for injection. The pH is preferably about 7.4. Generally, 40 milligrams of artelinate complexed with cyclodextrin per milliliter of solution is suitable. A dose of about 4-6 mg of artelinic acid (in complex) per kilogram of weight for a human is an appropriate dose. An injection of 10 ml of complex in solution or less is appropriate for treatment.
  • [0099]
    The formulation of the cyclodextrin complexed with artelinate in solution can be prepared and pumped through a filter into an injection vile, freeze dried for storage and later rehydrated with sterile water or saline or PBS for injection.
  • [0100]
    The cyclodextrin complexed with artelinate in solution can also be administered orally, sublingually, or in the form of a suppository.
  • [0000]
    Toxicity:
  • [0101]
    Cyclodextrins and artemisinins are both non-toxic to humans. However, large doses of cyclodextrins are not implicated in cases where kidneys are not fully functional.
  • [0000]
    In Vitro Data
  • [0102]
    In Vitro Inhibition Of Plasmodium Falciparum.
  • [0103]
    See U.S. Pat. No. 6,284,772, which is herein incorporated by reference. The in vitro assays were conducted by using a modification of the semiautomated mnicrodilution technique of Desjardins, et al. (1979) Antimicrob. Agents Chemther. 16:710-718 and Chulay et al. (1983) Exp. Parasitol. 55:138-146. Two strains of Plasmodium falciparum clones, from CDC Indochina III (W-2), CDC Sierra Leone I (D-6). The W-2 elone is susceptible to mefloquine but resistant to chloroquine, sulfadoxine, pyrimethamine, and quinine. The D-6 clone is resistant to mefloquine but susceptible to chloroquine, sulfadoxine, pyrimethamine, and quinine. These clones were derived by direct visualization and micromanipulation from patient isolates. Test compounds were initially dissolved in DMSO and diluted 400-fold in RPMI 1640 culture medium supplemented with 25 mM HEPES, 32 mM HaHCO3, and 10% Albumax I (GIBCO BRL, Grand Island, N.Y.). These solutions were subsequently serially diluted 2-fold with a Biomek 1000 (Beckrnan, Fullerton, Calif.) over 11 different concentrations. The parasites were exposed to serial dilutions of each compound for 48 h and incubated at 37° C. with 5% O2, 5% CO2, and 90% N2 prior to the addition of [3H]hypoxanthine. After a fuirther incubation of 18 h, parasite DNA was harvested from each microtiter well using Packard Filtermate 196 Harvester (Meriden, Conn.) onto glass filters. Uptake of [3H]hypoxanthine was measured with a Packard Topcount scintillation counter. Concentration-response data were analyzed by a nonlinear regression logistic dose-response model, and the IC50 values (50% inhibitory concentrations) for each compound were determined.
  • [0104]
    FIG. 28 indicates that both cyclodextrin formulations of artelinic acid β-cyclodextrin and hydroxypropyl-β-cyclodextrin) yielded very similar in vitro activity against multi-drug resistant strains of malaria as indicated. All data indicated IC50 concentrations within 4 ng/mL of the uncomplexed artelinate salt (artelinic acid control). Therefore, complexation of the artemisinin molecule was not found to inhibit antimalarial efficacy.
  • [0000]
    Advantages
  • [0105]
    The complexed cyclodextrin-artemisinins formulation does not precipitate or degrade over time. Formulations of artemisinins and cyclodextrin have been observed to remain completely soluble for up to seven weeks at elevated physiological temperatures (40 degrees C.) without any degradation and up to 6 months at room temperature. The complexed cyclodextrin formulation of the artemisinins does not change color over time. Formulations of artemisinins and cyclodextrin have been observed to remain colorless for several weeks at elevated physiological temperatures of 40 degrees C.
  • EXAMPLE Example 1 Formation of Artelinic Acid/Cyclodextrin Complex
  • [0106]
    Measure 2 moles of cyclodextrin and pre-issolve in buffer, deionized water, or saline. Sonicate the mixture to completely dissolve the cyclodextrin. Add 1 mole equivalent of artelinic acid and sonicate. Incubate at 40° C. for 2-3 hours. Higher concentrations of artelinic acid require longer incubation times, such as overnight, to promote complexation.
  • Example 2 Formation of Artesunic Acid/Cyclodextrin Complex
  • [0107]
    Measure 1 mole of cyclodextrin and pre-dissolve in buffer, deionized water, or saline. Sonicate the mixture to completely dissolve the cyclodextrin. Add 1 mole equivalent of artesunic acid and sonicate. Incubate at 40° C. for 2-3 hours. Higher concentrations of artesunic acid require longer incubation times to promote complexation.
  • [0108]
    The use of the complexed cyclodextrin formulation of the artemisinins described provides a shielding effect to protect the body from local toxic effects from the antimalarial agent until the drug is diluted sufficiently into the system.
  • [0109]
    The process of making the complexed artemisinins of the invention can be performed on a large scale using similar conditions.
  • [0110]
    Having now fully described the invention, it will be apparent to one of ordinary skill in the art that many changes and modifications can be made thereto without departing from the spirit or scope of the invention as set for the herein.

Claims (9)

  1. 1-57. (canceled)
  2. 58. A method of treating a patient having malaria: comprising administering to said patient a composition comprising a complexed cyclodextrin formulation of artemisinin, wherein said cyclodextrin is complexed with artelinic acid in a 2:1 molar ratio in aqueous solution.
  3. 59. The method of claim 58, wherein said administering is by intravenous injection, oral dose, sublingual dose, or suppository.
  4. 60. The method of claim 58, wherein said administering to said patient is by a dose of 4-6 milligrams of artelinic acid per kilogram of body weight.
  5. 61. The method of claim 58, wherein said 40 milligrams of said artemisinin complexed with cyclodextrin is dissolved per milliliter of aqueous solution.
  6. 62. A method of treating a patient with malaria: comprising administering to said patient an antimalarial composition comprising:
    a complexed cyclodextrin formulation of artemisinin, wherein said cyclodextrin is complexed with artesunic acid in a 1:1 ratio in an aqueous solution.
  7. 63. The method of claim 62, wherein said administering is by intravenous injection, oral dose, sublingual dose, or suppository.
  8. 64. The method of claim 62, wherein said administering to said patient is by a dose of 4-6 milligrams of artesunic acid per kilogram of body weight.
  9. 65. The method of claim 62, wherein said 40 milligrams of said artemisinin complexed with cyclodextrin is dissolved per milliliter of aqueous solution.
US11450009 2002-03-07 2006-06-09 Artemisinins with improved stability and bioavailability for therapeutic drug development and application Abandoned US20060229279A1 (en)

Priority Applications (4)

Application Number Priority Date Filing Date Title
US36298502 true 2002-03-07 2002-03-07
US10376387 US6951846B2 (en) 2002-03-07 2003-02-27 Artemisinins with improved stability and bioavailability for therapeutic drug development and application
US11113546 US7084132B2 (en) 2002-03-07 2005-04-25 Artemisinins with improved stability and bioavailability for therapeutic drug development and application
US11450009 US20060229279A1 (en) 2002-03-07 2006-06-09 Artemisinins with improved stability and bioavailability for therapeutic drug development and application

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
US11450009 US20060229279A1 (en) 2002-03-07 2006-06-09 Artemisinins with improved stability and bioavailability for therapeutic drug development and application

Publications (1)

Publication Number Publication Date
US20060229279A1 true true US20060229279A1 (en) 2006-10-12

Family

ID=27805252

Family Applications (3)

Application Number Title Priority Date Filing Date
US10376387 Expired - Fee Related US6951846B2 (en) 2002-03-07 2003-02-27 Artemisinins with improved stability and bioavailability for therapeutic drug development and application
US11113546 Expired - Fee Related US7084132B2 (en) 2002-03-07 2005-04-25 Artemisinins with improved stability and bioavailability for therapeutic drug development and application
US11450009 Abandoned US20060229279A1 (en) 2002-03-07 2006-06-09 Artemisinins with improved stability and bioavailability for therapeutic drug development and application

Family Applications Before (2)

Application Number Title Priority Date Filing Date
US10376387 Expired - Fee Related US6951846B2 (en) 2002-03-07 2003-02-27 Artemisinins with improved stability and bioavailability for therapeutic drug development and application
US11113546 Expired - Fee Related US7084132B2 (en) 2002-03-07 2005-04-25 Artemisinins with improved stability and bioavailability for therapeutic drug development and application

Country Status (2)

Country Link
US (3) US6951846B2 (en)
WO (1) WO2003075904A3 (en)

Families Citing this family (14)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8980290B2 (en) 2000-08-03 2015-03-17 Antares Pharma Ipl Ag Transdermal compositions for anticholinergic agents
ES2283425T3 (en) * 2000-08-03 2007-11-01 Antares Pharma Ipl Ag New composition for transdermal and / or transmucosal administration of active compounds which ensures adequate terapeutiocos levels.
US20040198706A1 (en) * 2003-03-11 2004-10-07 Carrara Dario Norberto R. Methods and formulations for transdermal or transmucosal application of active agents
WO2005039531A1 (en) 2003-10-10 2005-05-06 Antares Pharma Ipl Ag Transdermal pharmaceutical formulation for minimizing skin residues
WO2006125642B1 (en) 2005-05-27 2007-01-25 Antares Pharma Ipl Ag Methods and apparatus for transdermal or transmucosal application of testosterone
ES2367845T3 (en) * 2005-10-20 2011-11-10 Epipharm Gmbh Treatment and prevention of benign moles (naevi) artemisinin and its derivatives.
CA2646667C (en) 2006-04-21 2014-03-11 Antares Pharma Ipl Ag Methods of treating hot flashes with formulations for transdermal or transmucosal application
JP2009539989A (en) * 2006-06-13 2009-11-19 ウォルター リード アーミー インスティチュート オブ リサーチ (ダブリュアールエーアイアール) METHOD formulation and manufacture of injectable artesunic acid
GB0720967D0 (en) * 2007-10-25 2007-12-05 Protophama Ltd Anti-material pharmaceutical composition
WO2009100489A8 (en) * 2008-02-14 2011-10-06 Monash University Crystal structure of pfa-mi and the pfa-mi co4 complex
WO2010149215A1 (en) * 2009-06-25 2010-12-29 Dafra Pharma N.V. Artesunate pharmaceutical compositions soluble in aqueous solutions
WO2011009956A1 (en) * 2009-07-24 2011-01-27 Dafra Pharma N.V. Injectable aqueous solution containing artesunate
CN103864957B (en) * 2012-12-14 2016-03-16 昆药集团股份有限公司 Before drug artemisinin cyclodextrin and its preparation method of a carrier based on
CN103864962B (en) * 2012-12-14 2016-04-06 昆药集团股份有限公司 The method of preparing artemisinin prodrugs and cyclodextrin amine modified based

Citations (63)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4791135A (en) * 1987-08-20 1988-12-13 The United States Of America As Represented By The Secretary Of The Army Novel antimalarial dihydroartemisinin derivatives
US5001423A (en) * 1990-01-24 1991-03-19 International Business Machines Corporation Dry interface thermal chuck temperature control system for semiconductor wafer testing
US5220277A (en) * 1991-03-26 1993-06-15 Erich Reitinger Arrangement for testing semiconductor wafers or the like
US5266889A (en) * 1992-05-29 1993-11-30 Cascade Microtech, Inc. Wafer probe station with integrated environment control enclosure
US5802856A (en) * 1996-07-31 1998-09-08 Stanford University Multizone bake/chill thermal cycling module
US6407560B1 (en) * 1998-03-03 2002-06-18 Sandia Corporation Thermally-induced voltage alteration for analysis of microelectromechanical devices
US20020147177A1 (en) * 2001-01-24 2002-10-10 Yuen Kah Hay Formulation of artemisinin
US6549026B1 (en) * 1998-07-14 2003-04-15 Delta Design, Inc. Apparatus and method for temperature control of IC device during test
US6586464B2 (en) * 1999-01-12 2003-07-01 Johns Hopkins University Artemisinin analogs having antimalarial, antiproliferative, and antitumor activities and chemoselective methods of making the same
US6900652B2 (en) * 2003-06-13 2005-05-31 Solid State Measurements, Inc. Flexible membrane probe and method of use thereof
US6900653B2 (en) * 2002-07-05 2005-05-31 Samsung Electronics Co., Ltd. Needle fixture of a probe card in semiconductor inspection equipment and needle fixing method thereof
US6900647B2 (en) * 1996-05-23 2005-05-31 Genesis Technology Incorporated Contact probe and probe device
US6900646B2 (en) * 1998-04-03 2005-05-31 Hitachi, Ltd. Probing device and manufacturing method thereof, as well as testing apparatus and manufacturing method of semiconductor with use thereof
US6902941B2 (en) * 2003-03-11 2005-06-07 Taiwan Semiconductor Manufacturing Co., Ltd. Probing of device elements
US20050227503A1 (en) * 2002-04-15 2005-10-13 Erich Reitinger Method and device for conditioning semiconductor wafers and/or hybrids
US7002364B2 (en) * 2003-06-23 2006-02-21 Hynix Semiconductor Inc. Semiconductor device for reducing the number of probing pad used during wafer test and method for testing the same
US7001785B1 (en) * 2004-12-06 2006-02-21 Veeco Instruments, Inc. Capacitance probe for thin dielectric film characterization
US7002133B2 (en) * 2003-04-11 2006-02-21 Hewlett-Packard Development Company, L.P. Detecting one or more photons from their interactions with probe photons in a matter system
US7003184B2 (en) * 2000-09-07 2006-02-21 Optomed. As Fiber optic probes
US7002363B2 (en) * 2001-11-02 2006-02-21 Formfactor, Inc. Method and system for compensating thermally induced motion of probe cards
US7005842B2 (en) * 2000-12-22 2006-02-28 Tokyo Electron Limited Probe cartridge assembly and multi-probe assembly
US7005879B1 (en) * 2005-03-01 2006-02-28 International Business Machines Corporation Device for probe card power bus noise reduction
US7006046B2 (en) * 2001-02-15 2006-02-28 Integral Technologies, Inc. Low cost electronic probe devices manufactured from conductive loaded resin-based materials
US7005868B2 (en) * 2002-04-05 2006-02-28 Agilent Technologies, Inc. Apparatus and method for canceling DC errors and noise generated by ground shield current in a probe
US7009188B2 (en) * 2004-05-04 2006-03-07 Micron Technology, Inc. Lift-out probe having an extension tip, methods of making and using, and analytical instruments employing same
US7007380B2 (en) * 2000-07-13 2006-03-07 International Business Machines Corporation TFI probe I/O wrap test method
US7009415B2 (en) * 1999-10-06 2006-03-07 Tokyo Electron Limited Probing method and probing apparatus
US7009383B2 (en) * 1992-06-11 2006-03-07 Cascade Microtech, Inc. Wafer probe station having environment control enclosure
US7012425B2 (en) * 2003-09-18 2006-03-14 Tdk Corporation Eddy-current probe
US7012441B2 (en) * 2003-04-24 2006-03-14 Industrial Technology Research Institute High conducting thin-film nanoprobe card and its fabrication method
US7013221B1 (en) * 1999-07-16 2006-03-14 Rosetta Inpharmatics Llc Iterative probe design and detailed expression profiling with flexible in-situ synthesis arrays
US7011531B2 (en) * 2002-10-01 2006-03-14 International Business Machines Corporation Membrane probe with anchored elements
US7015703B2 (en) * 2003-08-12 2006-03-21 Scientific Systems Research Limited Radio frequency Langmuir probe
US7015707B2 (en) * 2002-03-20 2006-03-21 Gabe Cherian Micro probe
US7015708B2 (en) * 2003-07-11 2006-03-21 Gore Enterprise Holdings, Inc. Method and apparatus for a high frequency, impedance controlled probing device with flexible ground contacts
US7014499B2 (en) * 2004-07-05 2006-03-21 Yulim Hitech, Inc. Probe card for testing semiconductor device
US7015711B2 (en) * 2002-05-07 2006-03-21 Atg Test Systems Gmbh & Co. Kg Apparatus and method for the testing of circuit boards, and test probe for this apparatus and this method
US7015455B2 (en) * 1998-02-05 2006-03-21 Seiko Instruments Inc. Near-field optical probe
US7015689B2 (en) * 2002-12-19 2006-03-21 Sae Magnetics (H.K.) Ltd. Connection method for probe pins for measurement of characteristics of thin-film magnetic head and characteristic measurement method for thin-film magnetic head
US7015690B2 (en) * 2004-05-27 2006-03-21 General Electric Company Omnidirectional eddy current probe and inspection system
US7015709B2 (en) * 2004-05-12 2006-03-21 Delphi Technologies, Inc. Ultra-broadband differential voltage probes
US7019541B2 (en) * 2004-05-14 2006-03-28 Crown Products, Inc. Electric conductivity water probe
US7019544B1 (en) * 2001-12-21 2006-03-28 Lecroy Corporation Transmission line input structure test probe
US7020363B2 (en) * 2001-12-28 2006-03-28 Intel Corporation Optical probe for wafer testing
US7020360B2 (en) * 2001-11-13 2006-03-28 Advantest Corporation Wavelength dispersion probing system
US7022985B2 (en) * 2001-09-24 2006-04-04 Jpk Instruments Ag Apparatus and method for a scanning probe microscope
US7023226B2 (en) * 2003-02-20 2006-04-04 Octec Inc. Probe pins zero-point detecting method, and prober
US7023231B2 (en) * 2004-05-14 2006-04-04 Solid State Measurements, Inc. Work function controlled probe for measuring properties of a semiconductor wafer and method of use thereof
US7023225B2 (en) * 2003-04-16 2006-04-04 Lsi Logic Corporation Wafer-mounted micro-probing platform
US7023229B2 (en) * 2001-12-19 2006-04-04 Fujitsu Limited Dynamic burn-in equipment
US7022976B1 (en) * 2003-04-02 2006-04-04 Advanced Micro Devices, Inc. Dynamically adjustable probe tips
US7025628B2 (en) * 2003-08-13 2006-04-11 Agilent Technologies, Inc. Electronic probe extender
US7026834B2 (en) * 2003-06-24 2006-04-11 Agilent Technologies, Inc. Multiple two axis floating probe block assembly using split probe block
US7026832B2 (en) * 2002-10-28 2006-04-11 Dainippon Screen Mfg. Co., Ltd. Probe mark reading device and probe mark reading method
US7026833B2 (en) * 2000-08-24 2006-04-11 Texas Instruments Incorporated Multiple-chip probe and universal tester contact assemblage
US7026835B2 (en) * 1993-09-03 2006-04-11 Micron Technology, Inc. Engagement probe having a grouping of projecting apexes for engaging a conductive pad
US7030599B2 (en) * 2002-03-20 2006-04-18 Santronics, Inc. Hand held voltage detection probe
US7032307B2 (en) * 1999-12-21 2006-04-25 Kabushiki Kaisha Toshiba Method for fabricating a probe pin for testing electrical characteristics of an apparatus
US7034553B2 (en) * 2003-12-05 2006-04-25 Prodont, Inc. Direct resistance measurement corrosion probe
US7035738B2 (en) * 2001-05-29 2006-04-25 Hitachi Sofware Engineering Co., Ltd. Probe designing apparatus and probe designing method
US20060114012A1 (en) * 2004-11-26 2006-06-01 Erich Reitinger Method and apparatus for testing semiconductor wafers by means of a probe card
US20060158207A1 (en) * 2005-01-10 2006-07-20 Erich Reitinger Method and apparatus for testing semiconductor wafers by means of a temperature-regulated chuck device
US7101797B2 (en) * 2002-09-02 2006-09-05 Tokyo Electron Limited Substrate processing device and processing method

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE3346123A1 (en) * 1983-12-21 1985-06-27 Janssen Pharmaceutica Nv Pharmaceutical preparations of schwerloeslichen in water or unstable drugs and methods for their preparation
WO2001062299A3 (en) 2000-02-28 2002-01-31 Michael David Bentley Water-soluble polymer conjugates of artelinic acid

Patent Citations (65)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4791135A (en) * 1987-08-20 1988-12-13 The United States Of America As Represented By The Secretary Of The Army Novel antimalarial dihydroartemisinin derivatives
US5001423A (en) * 1990-01-24 1991-03-19 International Business Machines Corporation Dry interface thermal chuck temperature control system for semiconductor wafer testing
US5220277A (en) * 1991-03-26 1993-06-15 Erich Reitinger Arrangement for testing semiconductor wafers or the like
US5266889A (en) * 1992-05-29 1993-11-30 Cascade Microtech, Inc. Wafer probe station with integrated environment control enclosure
US7009383B2 (en) * 1992-06-11 2006-03-07 Cascade Microtech, Inc. Wafer probe station having environment control enclosure
US7026835B2 (en) * 1993-09-03 2006-04-11 Micron Technology, Inc. Engagement probe having a grouping of projecting apexes for engaging a conductive pad
US6900647B2 (en) * 1996-05-23 2005-05-31 Genesis Technology Incorporated Contact probe and probe device
US7015710B2 (en) * 1996-05-23 2006-03-21 Genesis Technology Incorporated Contact probe and probe device
US6903563B2 (en) * 1996-05-23 2005-06-07 Genesis Technology Incorporated Contact probe and probe device
US5802856A (en) * 1996-07-31 1998-09-08 Stanford University Multizone bake/chill thermal cycling module
US7015455B2 (en) * 1998-02-05 2006-03-21 Seiko Instruments Inc. Near-field optical probe
US6407560B1 (en) * 1998-03-03 2002-06-18 Sandia Corporation Thermally-induced voltage alteration for analysis of microelectromechanical devices
US6900646B2 (en) * 1998-04-03 2005-05-31 Hitachi, Ltd. Probing device and manufacturing method thereof, as well as testing apparatus and manufacturing method of semiconductor with use thereof
US6549026B1 (en) * 1998-07-14 2003-04-15 Delta Design, Inc. Apparatus and method for temperature control of IC device during test
US6586464B2 (en) * 1999-01-12 2003-07-01 Johns Hopkins University Artemisinin analogs having antimalarial, antiproliferative, and antitumor activities and chemoselective methods of making the same
US7013221B1 (en) * 1999-07-16 2006-03-14 Rosetta Inpharmatics Llc Iterative probe design and detailed expression profiling with flexible in-situ synthesis arrays
US7009415B2 (en) * 1999-10-06 2006-03-07 Tokyo Electron Limited Probing method and probing apparatus
US7032307B2 (en) * 1999-12-21 2006-04-25 Kabushiki Kaisha Toshiba Method for fabricating a probe pin for testing electrical characteristics of an apparatus
US7007380B2 (en) * 2000-07-13 2006-03-07 International Business Machines Corporation TFI probe I/O wrap test method
US7026833B2 (en) * 2000-08-24 2006-04-11 Texas Instruments Incorporated Multiple-chip probe and universal tester contact assemblage
US7003184B2 (en) * 2000-09-07 2006-02-21 Optomed. As Fiber optic probes
US7005842B2 (en) * 2000-12-22 2006-02-28 Tokyo Electron Limited Probe cartridge assembly and multi-probe assembly
US20020147177A1 (en) * 2001-01-24 2002-10-10 Yuen Kah Hay Formulation of artemisinin
US7006046B2 (en) * 2001-02-15 2006-02-28 Integral Technologies, Inc. Low cost electronic probe devices manufactured from conductive loaded resin-based materials
US7035738B2 (en) * 2001-05-29 2006-04-25 Hitachi Sofware Engineering Co., Ltd. Probe designing apparatus and probe designing method
US7022985B2 (en) * 2001-09-24 2006-04-04 Jpk Instruments Ag Apparatus and method for a scanning probe microscope
US7002363B2 (en) * 2001-11-02 2006-02-21 Formfactor, Inc. Method and system for compensating thermally induced motion of probe cards
US7020360B2 (en) * 2001-11-13 2006-03-28 Advantest Corporation Wavelength dispersion probing system
US7023229B2 (en) * 2001-12-19 2006-04-04 Fujitsu Limited Dynamic burn-in equipment
US7019544B1 (en) * 2001-12-21 2006-03-28 Lecroy Corporation Transmission line input structure test probe
US7020363B2 (en) * 2001-12-28 2006-03-28 Intel Corporation Optical probe for wafer testing
US7030599B2 (en) * 2002-03-20 2006-04-18 Santronics, Inc. Hand held voltage detection probe
US7015707B2 (en) * 2002-03-20 2006-03-21 Gabe Cherian Micro probe
US7005868B2 (en) * 2002-04-05 2006-02-28 Agilent Technologies, Inc. Apparatus and method for canceling DC errors and noise generated by ground shield current in a probe
US20050227503A1 (en) * 2002-04-15 2005-10-13 Erich Reitinger Method and device for conditioning semiconductor wafers and/or hybrids
US7015711B2 (en) * 2002-05-07 2006-03-21 Atg Test Systems Gmbh & Co. Kg Apparatus and method for the testing of circuit boards, and test probe for this apparatus and this method
US6900653B2 (en) * 2002-07-05 2005-05-31 Samsung Electronics Co., Ltd. Needle fixture of a probe card in semiconductor inspection equipment and needle fixing method thereof
US7101797B2 (en) * 2002-09-02 2006-09-05 Tokyo Electron Limited Substrate processing device and processing method
US7011531B2 (en) * 2002-10-01 2006-03-14 International Business Machines Corporation Membrane probe with anchored elements
US7026832B2 (en) * 2002-10-28 2006-04-11 Dainippon Screen Mfg. Co., Ltd. Probe mark reading device and probe mark reading method
US7015689B2 (en) * 2002-12-19 2006-03-21 Sae Magnetics (H.K.) Ltd. Connection method for probe pins for measurement of characteristics of thin-film magnetic head and characteristic measurement method for thin-film magnetic head
US7023226B2 (en) * 2003-02-20 2006-04-04 Octec Inc. Probe pins zero-point detecting method, and prober
US6902941B2 (en) * 2003-03-11 2005-06-07 Taiwan Semiconductor Manufacturing Co., Ltd. Probing of device elements
US7022976B1 (en) * 2003-04-02 2006-04-04 Advanced Micro Devices, Inc. Dynamically adjustable probe tips
US7002133B2 (en) * 2003-04-11 2006-02-21 Hewlett-Packard Development Company, L.P. Detecting one or more photons from their interactions with probe photons in a matter system
US7023225B2 (en) * 2003-04-16 2006-04-04 Lsi Logic Corporation Wafer-mounted micro-probing platform
US7012441B2 (en) * 2003-04-24 2006-03-14 Industrial Technology Research Institute High conducting thin-film nanoprobe card and its fabrication method
US6900652B2 (en) * 2003-06-13 2005-05-31 Solid State Measurements, Inc. Flexible membrane probe and method of use thereof
US7002364B2 (en) * 2003-06-23 2006-02-21 Hynix Semiconductor Inc. Semiconductor device for reducing the number of probing pad used during wafer test and method for testing the same
US7026834B2 (en) * 2003-06-24 2006-04-11 Agilent Technologies, Inc. Multiple two axis floating probe block assembly using split probe block
US7015708B2 (en) * 2003-07-11 2006-03-21 Gore Enterprise Holdings, Inc. Method and apparatus for a high frequency, impedance controlled probing device with flexible ground contacts
US7015703B2 (en) * 2003-08-12 2006-03-21 Scientific Systems Research Limited Radio frequency Langmuir probe
US7025628B2 (en) * 2003-08-13 2006-04-11 Agilent Technologies, Inc. Electronic probe extender
US7012425B2 (en) * 2003-09-18 2006-03-14 Tdk Corporation Eddy-current probe
US7034553B2 (en) * 2003-12-05 2006-04-25 Prodont, Inc. Direct resistance measurement corrosion probe
US7009188B2 (en) * 2004-05-04 2006-03-07 Micron Technology, Inc. Lift-out probe having an extension tip, methods of making and using, and analytical instruments employing same
US7015709B2 (en) * 2004-05-12 2006-03-21 Delphi Technologies, Inc. Ultra-broadband differential voltage probes
US7019541B2 (en) * 2004-05-14 2006-03-28 Crown Products, Inc. Electric conductivity water probe
US7023231B2 (en) * 2004-05-14 2006-04-04 Solid State Measurements, Inc. Work function controlled probe for measuring properties of a semiconductor wafer and method of use thereof
US7015690B2 (en) * 2004-05-27 2006-03-21 General Electric Company Omnidirectional eddy current probe and inspection system
US7014499B2 (en) * 2004-07-05 2006-03-21 Yulim Hitech, Inc. Probe card for testing semiconductor device
US20060114012A1 (en) * 2004-11-26 2006-06-01 Erich Reitinger Method and apparatus for testing semiconductor wafers by means of a probe card
US7001785B1 (en) * 2004-12-06 2006-02-21 Veeco Instruments, Inc. Capacitance probe for thin dielectric film characterization
US20060158207A1 (en) * 2005-01-10 2006-07-20 Erich Reitinger Method and apparatus for testing semiconductor wafers by means of a temperature-regulated chuck device
US7005879B1 (en) * 2005-03-01 2006-02-28 International Business Machines Corporation Device for probe card power bus noise reduction

Also Published As

Publication number Publication date Type
WO2003075904A2 (en) 2003-09-18 application
US20050187189A1 (en) 2005-08-25 application
US20030203875A1 (en) 2003-10-30 application
US6951846B2 (en) 2005-10-04 grant
US7084132B2 (en) 2006-08-01 grant
WO2003075904A3 (en) 2003-12-11 application

Similar Documents

Publication Publication Date Title
JonathanáBarnett et al. Catalysis by Cu 2+ of nitric oxide release from S-nitrosothiols (RSNO)
Ludueńa et al. Structure of the tubulin dimer.
Mergny et al. Fluorescence energy transfer as a probe for nucleic acid structures and sequences
Chidester et al. The structure of CC-1065, a potent antitumor agent and its binding to DNA
US4899755A (en) Hepatobiliary NMR contrast agents
US6028222A (en) Stable liquid paracetamol compositions, and method for preparing same
Bianchi et al. Thermodynamic and structural properties of Gd (III) complexes with polyamino-polycarboxylic ligands: basic compounds for the development of MRI contrast agents
US6476068B1 (en) Platinum derivative pharmaceutical formulations
Spielmann et al. Solution structure of a DNA complex with the fluorescent bis-intercalator TOTO determined by NMR spectroscopy
Kimura Macrocyclic polyamines as biological cation and anion complexones—An application to calculi dissolution
US5369101A (en) Expanded porphyrins: large porphyrin-like tripyrroledimethine-derived macrocycles
Beardsley et al. Studies of transfer RNA tertiary structure by singlet-singlet energy transfer
US5292414A (en) Expanded porphyrins: large porphyrin-like tripyrroledimethine-derived macrocycles for singlet oxygen production
US5256399A (en) Aromatic pentadentate expanded porphyrins in magnetic resonance imaging
Brewer et al. Conformational analysis of podophyllotoxin and its congeners. Structure-activity relationship in microtubule assembly
Maren et al. A new class of carbonic anhydrase inhibitor.
Cohen et al. Destruction of sympathetic nerve terminals by 6-hydroxydopamine: protection by 1-phenyl-3-(2-thiazolyl)-2-thiourea, diethyldithiocarbamate, methimazole, cysteamine, ethanol and n-butanol.
US4579849A (en) N-alkylguanine acyclonucleosides as antiviral agents
US6277990B1 (en) Substituted phenanthridinones and methods of use thereof
Stezowski Chemical-structural properties of tetracycline derivatives. 1. Molecular structure and conformation of the free base derivatives
Levine The physiological disposition of hexamethonium and related compounds
US6559154B2 (en) Composition of sodium channel blocking compound
Harvey et al. Multiple regions of metabolic activation of carcinogenic hydrocarbons
US4645661A (en) Method for alleviating cisplatin-induced nephrotoxicity and dithiocarbamate compounds for effecting same
Benoit-Vical et al. Trioxaquines are new antimalarial agents active on all erythrocytic forms, including gametocytes