BACKGROUND OF THE INVENTION

Herbal and botanical medicines have been used by people around the world for centuries. Today, herbal compositions, herbal extracts and compounds isolated from herbs play an important role in the treatment of a range of diseases and disorders and many of today's most commonly used medicines, such as aspirin, started as herbal remedies.

In order to reliably use botanical medicines for the treatment of disease, strict quality control is important. Unlike synthetic medicinal compositions, which can be more easily controlled and formulated, botanical medicines are derived from biologically derived plant matter. Each plant can contain a range of compounds in varied concentrations depending on genetic variation, growing conditions, the timing of planting and harvesting and the age of the plant matter when the botanical medicine is prepared.

Due to this variation, it is commonly observed that different batches of the same botanical medicine can have varied in vivo activity, depending on the diversity of the individual plants as well as variations in the preparation methods. It is also often observed that despite varied in vivo activity, different batches of an herbal medicine can still share very similar chemical profiles by standard chemical analysis methods such as chromatography and mass spectrometry. It is possible that slight impurities or compositional variations which cannot be detected through these chemical analysis methods can have a drastic effect on the overall activity of the botanical medicine. Thus, it is difficult to determine whether a specific batch of an herbal composition will have the desired in vivo activity simply through chemical analysis methods.

PHY906 is an herbal mixture extract based on the Huang Qin Tang herbal mixture, which was first described in Chinese texts 1,800 years ago, and has been used to treat gastrointestinal symptoms. PHY906 is composed of four herbs:Scutellaria baicalensis(S),Glycyrrhiza uralensis(G),Paeonia lactiflora(P), andZiziphus jujuba(Z). PHY906 is currently manufactured according to cGMP (current Good Manufacturing Practice). In clinical trials, PHY906 is shown to enhance the therapeutic index of chemotherapy against cancer.

There is thus an unmet need in the art for methods of determining the quality and potential efficacy of botanical medicines and other herbal compositions which do not rely solely on chemical analysis. The present invention satisfies this unmet need.

SUMMARY OF THE INVENTION

The present invention includes a method of evaluating the quality and potential in vivo activity of a test batch of an herbal composition, the method comprising subjecting the test batch of the herbal composition to one or more biological analysis methods selected from:

(a) a signaling transduction activity response assay; and

(b) a gene expression assay;

and then comparing the test batch results of the biological analysis method with results derived from a known batch of an herbal composition which has a known level of in vivo activity;

wherein a measurement of the difference between the test batch results and the known batch results provides an evaluation of the quality and potential in vivo activity of the test batch herbal composition.

In certain embodiments, the herbal composition comprises one or more compositions selected from the group consisting of herbal extracts ofScutellaria baicalensis(S),Glycyrrhiza uralensis(G),Paeonia lactiflora(P), andZiziphus jujuba(Z), any fractions thereof and any active chemicals present in the herbal extracts or fractions thereof. In other embodiments, the herbal composition is PHY906, wherein PHY906 comprises herbal extracts ofScutellaria baicalensis(S),Glycyrrhiza uralensis(G),Paeonia lactiflora(P), andZiziphus jujuba(Z) in a 3:2:2:2 (S:G:P:Z) ratio.

In certain embodiments, the signal transduction activity response assay comprises one or more assays selected from the group consisting of luciferase reporter assays and enzymatic assays.

In certain embodiments, the signal transduction activity response assay comprises measurement of signal transduction activity response against one or more enzymes selected from inflammation enzymes, cell growth and differentiation enzymes and endocrine and hormone enzymes. In other embodiments, the signal transduction activity response assay comprises measuring the signal transduction activity response against one or more signaling pathways selected from the group consisting of TNFa-NFkB, TLR2-NFkB, TLR4-NFkB, IL6-stat3, IFNg-stat1/1, IFNa-stat1/2, DEX-GR, COX-2, iNOS, NRF2, TGFb-Smad2/3, TPA-AP1, CREB, wnt3a-Lef/b-cat, VD3-VDR, ER-alpha, ER-beta, DHT-AR and aldosterone-MR.

In certain embodiments, the gene expression assay comprises: treating HepG2 cells with the herbal composition for 24 h, extracting the mRNA produced and quantifying the mRNA through qRT-PCR analysis.

In certain embodiments, the gene expression assay comprises measurement of one or more genes encoding proteins having a function selected from inflammation, anti-oxidation, growth and differentiation, metabolism, and cell-cell interaction. In other embodiments, the one or more genes are selected from the group consisting of ICAM, IRF5, AKR1C1, HO1, GCLC, GCLM, Axin2, GDF15, IGFBP3, OKL38, PIM1, SERTAD, SOS1, BHMT2, CPT1A, SLC7A11, CD24, EMP2 and KRT23.

In certain embodiments, the one or more biological analysis methods differentiate between active batches of the herbal composition and inactive batches of the herbal composition better than chemical composition analysis methods. In other embodiments, the chemical composition analysis methods include LC-MS (liquid chromatography-mass spectrometry).

In certain embodiments, if the test batch results are between about 90% and about 110% of the known batch results then the test batch is determined to have sufficiently similar quality as the known batch and potential in vivo activity.

In certain embodiments, the quality of the test batch of the herbal composition is further analyzed by LC-MS.

The invention further provides a method of treating a subject with cancer, the method comprising evaluating the quality and potential in vivo efficacy of a test batch of PHY906 by subjecting the herbal composition to one or more biological analysis methods selected from:

(a) a signaling transduction activity response assay; and

(b) a gene expression assay;

comparing the results of the biological analysis method with results derived from a known batch of PHY906 which has a known level of in vivo activity;

wherein a measurement of the difference between the test batch results and the known batch results provides an evaluation of the quality and potential in vivo activity of the test batch herbal composition,

and if the test batch of PHY906 shows similar activity in the biological analysis methods to the known batch of PHY906, administer the test batch of PHY906 to the subject.

