This plant offers unique medicinal qualities, particularly when treating fungal infections, which are often overlooked, and, I think, is a prime pathogen that causes bleeding in the racehorse. Osage orange is also relatively non-toxic unlike many antifungal prescriptions. For those with cancer, it has cancer fighting properties as well.

Osage Orange (Maclura pomifera) is a tree/shrub of unending uniqueness and use. Growing up on a Midwestern farm, it is my sacred tree. I say tree, because if left to its natural instincts this once propagated livestock hedge will grow into 30' trees. This tree has always fascinated me as no other tree could. The wood can easily last 50 years without any appreciable decay. A log will sink if thrown into a pond. It will burn the hottest of any wood in the Midwest and in addition, burn green. The hardness and durability of the Osage wood is unparalleled. Before barb-wire, this tree was sold throughout the Midwest as a livestock hedge fence which in four years was said to grow to be, horse-high, bull-strong, and hog-tight.

     I have always been interested in natural anti-microbials found in nature. I have been much impressed with the Olive tree leaf as a very effective anti-microbial and often wished that a similar tree grew in my area. The Olive tree is very disease resistant, few insects seem to bother it, and it can live for an eternity. I was also equally impressed with the South American tree, Pau d'Arco. The Pau d'Arco is widely coveted as a superb anti-fungal medicinal and it is a specific for leukemia. In fact, it may have been instrumental in saving my brother who succumbed to leukemia over 5 years ago. I placed him on Pau d'Arco and I think it might have helped him survive! It eventually occurred to me that the physical descriptions of the Olive and Pau d'Arco trees also described our locally growing American Osage Orange tree. The Maclura pomifera never seems to die. Its leaves are as bright green and perfect from insect or microbe damage from Spring to Fall's end. Its wood, dead or alive, seems almost impervious to all fungi that will attack it. Few dead woods have that property unprotected out in the open! The Osage Orange is a vibrant life force. But best of all, the Osage Orange leaves are non-toxic. I have seen livestock routinely take mouthfuls from the lower branches without ill effect. Perhaps they were even doctoring themselves?

Description:    Osage Orange is a deciduous tree which can grow beyond 30 feet. It has a rounded silhouette with the main trunk generally rather short. The roundness is due to many radiating large branches. The twigs tend to zigzag with leaves that alternate. Leaves are simple, oval affairs with a long stem and sharp, long point. They are smooth edged, a glossy dark-green on top, a dull light green on the bottom, and have a characteristic milky sap excreted from torn petioles. Leaves tend to be from 3 to 5 inches in length. Some varieties produce thorns, others don't. The famous fruit, hedge apples, are only produced by female trees. The fruit resembles oranges with brain-like outer skin. The fruit of female trees must be pollinated in order for the fruit to contain seeds.

Location:     Osage Orange was originally native to a very limited belt starting in Southwestern Arkansas and neighboring Southeastern Oklahoma southward into Eastern Texas and Northwestern Louisiana. Now through manual commerce and introduction, this tree can be found in almost all sections of the USA. It is a very drought tolerant tree, easily transplanted, and tolerates poor soils, extreme heat, and occasional subzero cold spells well.

Parts Used: Roots, inner bark, and leaves.

Medicinal Properties:    It is said that the Indians utilized a root/water infusion for eye conditions, but not much more seemed to have been done medicinally with this tree from an ethnobotanical point of view. If the truth be known, I would bet this tree had far more native American applications medicinally than the white man will ever be aware of. The leaves and inner bark will probably offer the most promise for future medicinal compound extractions—not so much the roots or the fruit. Modern researchers are scrutinizing many of the compounds in the Osage Orange. One of the active compounds is Tetrahydroxystilbene (THS) which shows significant antifungal activity and probably the one we are most interested in. It is also known as an analog of Resveratrol. That name should ring a bell with most of you! Resveratrol (3,5,4'-trihydroxy-trans-stibene), a phytoalexin present in grapes, peanuts and pines has antioxidant and anti-inflammatory activities. The TV show, 60 Minutes did a piece on this exciting new compound as featured in wines a few years ago and set off the craze. However, it should be noted that no one herb or tree is simply one isolated compound! An herb is a sum of its many parts and the Osage Orange tree should be considered in that light. You can take out of it tetrahydroxystilbene but there are far too many other synergistic compounds to only relegate this tree to just a tetrahydroxystilbene source. It was found in a study below that tetrahydroxystilbene was not very effective against combating the common fungus, Candida albicans. I guarantee one thing! The Osage Orange is very resistant to Candida infections! So there is much more to this tree than just tetrahydroxystilbene! The Osage Orange is a soup of many anti-microbial chemicals working together to protect this tree in a way that few trees have ever been protected. It is just not about tetrahydroxystilbene, but because so much research has been done on this one compound, we will concentrate on it below.

      There are a number of herbalists and alternative health researchers that will tell you that fungi in its many forms is the true general cause of cancer and other chronic diseases. It may or may not be, but I am sure of one thing, fungi is absolutely underestimated by the medical community in the havoc it can cause in both human and animal pathology. There are few really good, nontoxic antifungals out there for one to use. Osage Orange is one plant that can bring you a very effective non-toxic, antifungal component. There isn't much out there on the Internet on the value of Osage Orange for medicinal purposes. You will run across some people using the hedge apple fruit as the main part of the tree to fight cancer, etc. I doubt seriously they are doing themselves justice by concentrating on just the fruit. As with most medicinal trees, the value is not in the fruit, but found in the leaf or the inner bark of trees. Osage Orange is no different! Almost without exception in the herbal literature, the inner bark (cambium layer and phloem) contains the most powerful concentration of a plant's chemicals. Compare the Osage to the Olive tree. The olive fruit is unpalatable in its natural state just like the Osage orange ball and both are not the part of the tree generally used in herbal medicine as a remedy. If you want to experiment with Osage Orange, I urge you to forego the fruit in favor of the leaf or inner bark. If some people are getting that much benefit from just eating the raw fruit, think what the potent leaf or inner bark will do medicinally! I would suggest either the ethanol extract for oral use or the DMSO extract for topical application in your animals.

There has been quite a bit of research done on Tetrahydroxystilbene as a tumor inhibitor. Here we find this wonderful analog in the Osage Orange!

Here is an interesting study on Tetrahydroxystilbene as an antifungal:


http://www.scienceasia.org/1976.02.n4/v02_202_205.pdf

Other promising antibiotic compounds have been isolated as a non-toxic food preservative from the osage orange. The subfraction with the greatest inhibitory effect on wood decay fungi as characterized by spectroscopic methods, seems to be a mixture of tetra- & pentahydroxystilbenes. Research is continuing in the laboratory and there is no reason why we can not do our own veterinary experimenting. Tetrahydroxystilbene may not be the only healing component to this unique tree! Osage Orange extracts should offer a very interesting non-toxic alternative to combating various infections of a fungal and bacterial nature.

Here is an interesting study on Tetrahydroxystilbene as a tumor inhibitor:

http://cancerres.aacrjournals.org/content/61/22/8164.full

And the role of Tetrahydroxystilbene in the treatment of cancer:

http://www.createbalance.net/support/science/resveratrol_cancer_prevention.pdf

2001 Abstract:

Resveratrol, a trihydroxystilbene found in grapes and other plants, has been shown to be active in inhibiting multistage carcinogenesis. Using resveratrol as a prototype, we have synthesized a number of polyhydroxy- and polymethoxy-stilbenes and tested their anti-proliferative effect in normal and transformed human cells. Here we show that one of the resveratrol analogs, 3,4,5,4'-tetrahydroxystilbene (R-4), specifically inhibited the growth of SV40 virally transformed WI38 cells (WI38VA) at 10 ¥M, but had no effect on normal WI38 cells at even higher concentrations. R-4 also prominently induced apoptosis in WI38VA cells, but not in WI38 cells. RNase protection assay showed that R-4 significantly induced the expression of p53, GADD45 and Bax genes and concomitantly suppressed the expression of bcl-2 gene in WI38VA, but not in WI38 cells. A large increase in p53 DNA binding activity and the presence of p53 in the Bax promoter binding complex suggested that p53 was responsible for the Bax gene expression induced by R-4 in transformed cells. Within 4 h of treatment with R-4, the Bax to bcl-2 protein ratio in WI38 and WI38VA cells was, respectively, 0.1 and 105, a difference of three orders of magnitude. While R-4 prominently induced the p53/Bax pro-apoptotic genes, it also concomitantly suppressed the expression of Cox-2 in WI38VA cells. Taken together, our study suggests that the induction of p53 gene by R-4 in transformed cells may play a key role in the differential growth inhibition and apoptosis of transformed cells.

A 2008 study on Human Protrate Cancer cells:

Induction of cell cycle arrest in DU145 human prostate cancer cells by the dietary compound piceatannol Piceatannol (3,4,3',5'-tetrahydroxystilbene) is a naturally occurring polyphenol and analog of the cancer chemopreventive agent resveratrol (3,5,4'-trihydroxystilbene). Piceatannol is present in the skins of grapes, rhubarb, and sugar cane and is known to have anti-inflammatory and anti-carcinogenic activity. In the present study, we examined piceatannol-mediated regulation of cell cycle progression in androgen-insensitive DU145 human prostate cancer cells. Cells were treated with 0-10 µM piceatannol. The percentage of cells in G1 phase was increased in cells treated with piceatannol for 24 h. Piceatannol decreased incorporation of [3H]thymidine into DNA. Piceatannol decreased the protein levels of cyclin A, and cyclin D1. In addition, cyclin-dependent kinase (CDK)2 and CDK4 protein levels decreased in cell treated with piceatannol. But piceatannol did not affect the expression of the CDK inhibitor p21 or p27. The activity of CDK2 and CDK4 decreased following piceatannol treatment in a concentration-dependent manner. The present results indicate that piceatannol inhibits the growth of DU145 cell growth by delaying G1 cell cycle progression, which may be mediated via inhibition of CDK activities.

The Osage Orange Heartwood:

Oxyresveratrol (trans-2,4,3',5'-tetrahydroxystilbene)

Abstract: Analysis of osage orange (Maclura pomifera) heartwood extractives showed that two compounds were present at much higher levels than previously reported, the flavanonol dihydromorin (2.51%) and the stilbene oxyresveratrol (2.65%). All compounds present in osage orange heartwood that were tested had low activity against wood-decaying fungi. The agar plate test showed no synergistic effect, but the soil block test using white-rot fungi suggested synergism. The high durability of osage orange heartwood may be due to the large amounts of oxyresveratrol and dihydromorin. High levels of one or two monomeric compounds may also explain the exceptionally good durability of black locust and red mulberry heartwood.

The amount of stilbenes that occurs in wine is far less than the amount found in the heartwood of Osage orange, Maclura pomifera. The potential use of Maclura pomifera in the industry as a medicinal crop may be an advantage due to its growing throughout the region and its stilbene content of 1.96% in the heartwood. According to some data, in the living organisms oxyveratrol is converted to its isomer with catohol structure.

Abstract: 1,3,6,7-Tetrahydroxyxanthone has been isolated from the heartwood of the osage orange ( Maclura pomifera Raf.). The occurrence in this wood of morin and of 2,3¥,4,5¥-tetrahydroxystilbene has been confirmed.

Abstract: Oxidative stress is one of the major pathological factors in the cascade that leads to cell death in cerebral ischemia. Here, we investigated the neuroprotective effect of a naturally occurring antioxidant, oxyresveratrol, to reduce brain injury after cerebral stroke. We used the transient rat middle cerebral artery occlusion (MCAO) model of brain ischemia to induce a defined brain infarction. Oxyresveratrol was given twice intraperitoneally: immediately after occlusion and at the time of reperfusion. Oxyresveratrol (10 or 20 mg/kg) significantly reduced the brain infarct volume by approximately 54% and 63%, respectively, when compared to vehicle-treated MCAO rats. Also, the neurological deficits as assessed by different scoring methods improved in oxyresveratrol-treated MCAO rats. Histological analysis of apoptotic markers in the ischemic brain area revealed that oxyresveratrol treatment diminished cytochrome c release and decreased caspase-3 activation in MCAO rats. Also, staining for apoptotic DNA showed that the number of apoptotic nuclei in ischemic brain was reduced after oxyresveratrol treatment as compared to the vehicle-treated MCAO rats. This dose-dependent neuroprotective effect of oxyresveratrol in an in vivo stroke model demonstrates that this drug may prove to be beneficial for a therapeutic strategy to limit brain injury in acute brain ischemia.

THE CONSUMPTION OF NUTRIENTS rich in hydroxystilbenes (resveratrol and oxyresveratrol), such as grapes, red wine, peanuts and mulberry wood extracts (OSAGE ORANGE), has beneficial effects on human health (8, 9, 56). The cellular and the molecular mechanisms that underlie these beneficial effects are largely unknown. Several in vitro studies suggest that resveratrol suppresses the expression of genes involved in the inflammatory response to bacterial and viral stimuli (41, 57). More specifically, resveratrol or its hydroxylated form, oxyresveratrol (o-RES; trans-2,3',4,5'-tetrahydroxystilbene), reduces the immune activation of the inducible form of cyclooxygenase (COX-2), a key enzyme in the production of inflammatory prostaglandins (31, 38, 51). Such an effect is likely due to the resveratrol-induced inhibition of the NF-B signaling pathway, a signaling pathway involved in cox-2 gene expression (26, 30). However, the inflammatory responses in vivo are much more complex than can be modeled in cultured cells in vitro. As a consequence, there are conflicting data on the in vitro and in vivo effects of resveratrol (16), most likely because in vivo immune system activation sets in motion complex and intricate inflammatory responses not seen in vitro. For example, in the best-studied model of innate immune system activation (i.e., a systemic injection of LPS, a component of the outer coat of the gram-negative bac teria), LPS activates resident macrophages in a variety of tissues (17, 28, 45) to release proinflammatory cytokines such as tumor necrosis factor (TNF)-¥, IL-1, and IL-6, as well as anti-inflammatory cytokines and hormones with anti-inflammatory activity (12). A variety of transcription factors such as NF-¥B and STAT-3 are consequently activated and lead to the induction of COX-2 in a variety of tissues in the body, including the brain. One hallmark of innate immune system activation in vivo is fever, an important component of the host defense response against infection (22). At the molecular level, fever develops as a result of COX-2 induction in endothelial cells of the brain vasculature followed by a subsequent production of prostaglandin E2 (PGE2) (5, 32). PGE2 induces fever by acting on neurons located within organum vasculosum of the lateral terminalis (OVLT) and the ventral preoptic region of the hypothalamus (49, 50). In addition to fever, LPS, at higher doses, can also induce hypothermia. It is we ll established that LPS-induced hypothermia is mediated by the proinflammatory cytokine TNF-¥ (10, 11, 27, 47). Hydroxystilbenes are available as nonprescription, complementary medications with largely undocumented claims of efficacy. It is important that the actions of such compounds be subjected to rigorous experimental assessment, so that both the accuracy of claims and the action of these nonproprietary medications are documented. The LPS fever model provides such an opportunity in light of its well-understood and characterized actions. Thus one objective of this study was to determine whether o-RES is able to alter LPS fever via an action on LPS-induced COX-2 in rat OVLT and preoptic area (OVLT/POA). We further explored whether o-RES alters levels of LPS-stimulated, blood-born inflammatory cytokines, such as TNF-¥ and IL-6 and the level of activation of the transcription factor NF-¥B in the liver, one major target for bacterial LPS (28).

Abstract:

Hydroxystilbenes are naturally occurring polyphenols with protective effects against reactive oxygen and nitrogen species (ROS/RNS). Here, we investigated oxyresveratrol (OXY), which is contained in high amounts in mulberry wood (osage orange), in comparison to the antioxidant resveratrol (RES). We found that OXY is a more effective scavenger for 2,2-diphenyl-1-picryl-hydrazyl (DPPH, 100 ¥M) used as a general free radical model, compared to RES or trans-4-hydroxystilbene(IC50 =28.9, 38.5, and 39.6 ¥M, respectively). When primary glial cell cultures were loaded with the ROS/RNS-sensitive fluorochrome 2,7-dichlorodihydrofluorescein, the lowest rise in the fluorescence signal after H2O2 exposure was seen when the cells were pretreated with OXY. Using 4,5-diaminofluorescein (DAF-2) to monitor free nitric oxide levels (7.7 ¥M NO) in a spectrofluorimetric cell-free assay, we found again that OXY (at 5 ¥M) is a more effective scavenger. Accordingly, cultures of the murine microglial cell line N9 and primary mixed glial cultures were used to test the drug effects of NO production upon expression of the inducible isoform of nitric oxide synthase (iNOS). We found that both compounds considerably diminished NO (nitrite) levels, RES more effectively than OXY (IC50 =22.36 and 45.31 ¥M). RES but not OXY down-regulated the expression of iNOS protein, but both did not alter iNOS activity.Furthermore, OXY displayed a generally lower cytotoxicity than RES. The radical and ROS scavenging properties, as well as the lower cytotoxicity towards microglia and the known good water solubility suggest OXY as a potential protectant against ROS/RNS.

Tyrosinase is responsible for the molting process in insects, undesirable browning of fruits and vegetables, and coloring of skin, hair, and eyes in animals. To clarify the mechanism of the depigmenting property of hydroxystilbene compounds, inhibitory actions of oxyresveratrol and its analogs on tyrosinases from mushroom and murine melanoma B-16 have been elucidated in this study. Oxyresveratrol showed potent inhibitory effect with an IC(50) value of 1.2 microm on mushroom tyrosinase activity, which was 32-fold stronger inhibition than kojic acid, a depigmenting agent used as the cosmetic material with skin-whitening effect and the medical agent for hyperpigmentation disorders. Hydroxystilbene compounds of resveratrol, 3,5-dihydroxy-4'-methoxystilbene, and rhapontigenin also showed more than 50% inhibition at 100 microm on mushroom tyrosinase activity, but other methylated or glycosylated hydroxystilbenes of 3,4'-dimethoxy-5-hydroxystilbene, trimethylresveratrol, piceid, and rhaponticin did not inhibit significantly. None of the hydroxystilbene compounds except oxyresveratrol exhibited more than 50% inhibition at 100 microm on l-tyrosine oxidation by murine tyrosinase activity; oxyresveratrol showed an IC(50) value of 52.7 microm on the enzyme activity. The kinetics and mechanism for inhibition of mushroom tyrosinase exhibited the reversibility of oxyresveratrol as a noncompetitive inhibitor with l-tyrosine as the substrate. The interaction between oxyresveratrol and tyrosinase exhibited a high affinity reflected in a K(i) value of 3.2-4.2 x 10(-7) m. Oxyresveratrol did not affect the promoter activity of the tyrosinase gene in murine melanoma B-16 at 10 and 100 microm. Therefore, the depigmenting effect of oxyresveratrol works through reversible inhibition of tyrosinase activity rather than suppression of the expression and synthesis of the enzyme. The number and position of hydroxy substituents seem to play an important role in the inhibitory effects of hydroxystilbene compounds on tyrosinase activity.

Abstract: The anti-herpes simplex virus (HSV) compound, oxyresveratrol, purified from a Thai traditional medicinal plant of Artocarpus lakoocha, was evaluated for its anti-varicella-zoste r virus (VZV) activity. This compound exhibited IC(50) values (50%-inhibitory concentrations for virus plaque formation) of 12.82, 12.80, 12.99 and 12.82 microg/ml against wild type, thymidine kinase-deficient and two types of DNA polymerase mutants with acyclovir-resistance , respectively. Thus oxyresveratrol showed a broad spectrum of anti-VZV activity with a mechanism of action different from that of acyclovir.

Oxyresveratrol (OXY) is a polyhydroxylated stilbene existing in mulberry (osage orange). Increasing lines of evidence have shown its neuroprotective effects against Alzheimer disease and stroke. However, little is known about its neuroprotective effect in Parkinson disease (PD). Owing to its antioxidant activity, blood-brain barrier permeativity, and water solubility, we hypothesized that OXY may exert neuroprotective effects against parkinsonian mimetic 6-hydroxydopamine (6-OHDA) neurotoxicity. Neuroblastoma SH-SY5Y cells have long been used as dopaminergic neurons in PD research. We found that both pretreatment and posttreatment with OXY on SH-SY5Y cells significantly reduced the release of lactate dehydrogenase, the activity of caspase-3, and the generation of intracellular reactive oxygen species triggered by 6-OHDA. Compared to resveratrol, OXY exhibited a wider effective dosage range. We proved that OXY could penetrate the cell membrane by HPLC analysis of cell extracts. These results suggest that OXY may act as an intracellular antioxidant to reduce oxidative stress induced by 6-OHDA. Western blot analysis demonstrated that OXY markedly attenuated 6-OHDA-induced phosphorylation of JNK and c-Jun. Furthermore, we proved that OXY increased the basal levels of SIRT1, which may disclose new pathways accounting for the neuroprotective effects of OXY. Taken together, our results suggest OXY, a dietary phenolic compound, as a potential nutritional candidate for protection against neurodegeneration in PD.

Abstract: Hydroxystilbenes are naturally occurring polyphenols with protective effects against reactive oxygen and nitrogen species (ROS/RNS). Here, we investigated oxyresveratrol(OXY), which is contained in high amounts in mulberry wood, (osage orange) in comparison to the antioxidant resveratrol (RES). We found that OXY is a more effective scavenger for 2,2-diphenyl-1-picryl-hydrazyl (DPPH, 100¥M) used as a general free radical model, compared to RES or trans-4-hydroxystilbene (IC50=28.9, 38.5, and 39.6¥M, respectively). When primary glial cell cultures were loaded with the ROS/RNS-sensitive fluorochrome 2,7-dichlorodihydrofluorescein, the lowest rise in the fluorescence signal after H2O2 exposure was seen when the cells were pretreated with OXY. Using 4,5-diaminofluorescein (DAF-2) to monitor free nitric oxide levels (7.7 ¥M NO) in a spectrofluorimetric cell-free assay, we found again that OXY (at 5¥M) is a more effective scavenger. Accordingly, cultures of the murine microglial cell line N9 and primary mixed glial cultures were used to test the drug effects of NO production upon expression of the inducible isoform of nitric oxide synthase (iNOS). We found that both compounds considerably diminished NO (nitrite) levels, RES more effectively than OXY (IC50=22.36 and 45.31¥M). RES but not OXY down-regulated the expression of iNOS protein, but both did not alter iNOS activity. Furthermore, OXY displayed a generally lower cytotoxicity than RES. The radical and ROS scavenging properties, as well as the lower cytotoxicity towards microglia and the known good water solubility suggest OXY as a potential protectant against ROS/RNS.

The Osage Orange Fruit:

Anti-inflammatory and antinociceptive potential of Maclura pomifera fruit:

Abstract: The aqueous, ethanolic and chloroform extracts and two prenylated isoflavones: scandenone (I) and auriculasin (II), isolated from the fruits of Maclura pomifera (Rafin.) Schneider, were investigated for their in vivo anti-inflammatory and antinociceptive activity. For the anti-inflammatory activity, both carrageenan-induced hind paw edema and 12-O-tetradecanoyl-13-acetate (TPA)-induced mouse ear edema models and for the antinociceptive activity, p-benzoquinone-induced abdominal constriction test were used. Scandenone, the chloroform and the ethanolic extracts were shown to possess antinociceptive activity and anti-inflammatory activity on carrageenan-induced hind paw edema model at 100 mg/kg dose. The same compound and the extract were also found to be highly active in (TPA)-induced mouse ear edema model whereas auriculasin and the H2O extract showed to be inactive in all of the assays.

Abstract: Flavonoids and isoflavonoids are well known for their beneficial effects on human health and their anti-insect and anti-microbial activities in plants. Osage orange fruit is rich in prenylated isoflavones and dihydrokaempferol and its glucoside. Four glycosyltransferases were identified from a collection of osage orange fruit expressed sequence tags. Biochemical characterization suggested that the glycosyltransferase UGT75L4 might be responsible for glucosylation of dihydrokaempferol in vivo, although this enzyme exhibited broad substrate recognition toward isoflavonoids and flavonoids in vitro. UGT88A4 was active on coumarin substrates. Identification of highly active phenylpropanoid glycosyltransferases will facilitate the metabolic engineering of glycosylated natural products in plants.

Abstract: The major constituents of fruits of Maclura pomifera are the prenylated isoflavones, osajin (1) and pomiferin (2). Since significant biological activities of extracts from the wood of M. pomifera were previously reported, the peroxynitrite scavenging activity, inhibition of lipid peroxidation, scavenging of DPPH and EROD activity of these two major substances were studied.

Abstract: This study investigated the isoflavone content in a unique nonedible tree fruit, Osage orange [Maclura pomifera (Raf.) Schneid], and methods for the extraction, identification, and quantification of the two major isoflavones, osajin and pomiferin, were developed. The ethyl acetate extract contained 25.7% osajin and 36.2% pomiferin, and the two isoflavones were at 9.5 g kg-1 of fresh Osage orange. Two model systems, FRAP and ¥-CLAMS, were used to measure the antioxidant activity of these two isoflavones. Pomiferin was found to be a strong antioxidant in both systems, comparable to the antioxidant vitamins C and E and the synthetic antioxidant BHT. Osajin and the two soybean isoflavones (genistein and daidzein) showed no antioxidant activity. Although the Osage orange fruit is not a food source, it is considered to be safe and, therefore, a potentially good source of an antioxidant nutraceutical and functional food ingredient.

Abstract: A reinvestigation of the constituents of the Osage orange (Maclura pomifera ) yielded, in addition to the previously reported triterpenes (lupeol, butyrospermol, and lupane-3¥,20-diol), the pigments osajin and pomiferin, and a previously unreported constituent. The structure of this new compound was investigated. On the basis of spectroscopic and chemical data, it appeared to be an epimer of lupeol and is referred to as 19-H-lupeol.

Lupeol as found in the osage orange has been shown of value in Pancreatic Cancer:

Abstract: Pancreatic cancer is an exceptionally aggressive disease, the treatment of which has largely been unsuccessful due to higher resistance offered by pancreatic cancer cells to conventional approaches such as surgery, radiation and/or chemotherapy. The aberration of Ras oncoprotein has been linked to the induction of multiple signaling pathways and to the resistance offered by pancreatic cancer cells to apoptosis. Therefore, there is a need for development of new and effective chemotherapeutic agents which can target multiple pathways to induce responsiveness of pancreatic cancer cells to death signals. In this study, human pancreatic adenocarcinoma cells AsPC-1 were used to investigate the effect of Lupeol on cell growth and its effects on the modulation of multiple Ras-induced signaling pathways. Lupeol caused a dose-dependent inhibition of cell growth as assessed by MTT assay and induction of apoptosis as assessed by flow cytometry, fluorescence microscopy and western blotting. Lupeol treatment to cells was found to significantly reduce the expression of Ras oncoprotein and modulate the protein expression of various signaling molecules involved in PKC¥a/ODC, PI3K/Akt and MAPKs pathways along with a significant reduction in the activation of NF¥?B signaling pathway. Our data suggest that Lupeol can adopt a multi-prong strategy to target multiple signaling pathways leading to induction of apoptosis and inhibition of growth of pancreatic cancer cells. Lupeol could be a potential agent against pancreatic cancer, however, further in-depth in vivo studies are warranted.

Researchers induced head and neck cancer in mice and then gave the mice doses of the chemical lupeol, a triterpene compound found in many fruits. The lupeol appeared to block the function of a protein called NFkB, which assists both healthy and malignant cells in the process of growth and repair. In addition, the growth and spread of the cancer appeared to be slowed. From the animal models, not only did lupeol suppress the spread, the tumor got smaller. Compared to conventional drugs, lupeol reduced the size of the tumor far faster," researcher Terence Lee said.

Abstract: In prostate cancer, a fine balance between cell proliferation and apoptotic death is lost, resulting in increased cellular mass and tumor progression. One approach to redress this imbalance and control this malignancy is its preventive intervention through the use of dietary natural agents. Here, we investigated the growth-inhibitory effect and associated mechanisms of Lupeol, a triterpene present in fruits and vegetables, in androgen-sensitive human prostate cancer cells. Lupeol treatment resulted in significant inhibition of cell viability in a dose-dependent manner and caused apoptotic death of prostate cancer cells. Lupeol was found to induce the cleavage of poly(ADP-ribose) polymerase protein and degradation of acinus protein with a significant increase in the expression of FADD protein. Among all death receptor targets examined, Lupeol specifically caused a significant increase in the expression of Fas receptor. The small interfering RNA-mediated silencing of the Fas gene and inhibition of caspase-6, caspase-8, and caspase-9 by their specific inhibitors confirmed that Lupeol specifically activates the Fas receptor-mediated apoptotic pathway in androgen-sensitive prostate cancer cells. The treatment of cells with a combination of anti-Fas monoclonal antibody and Lupeol resulted in higher cell death compared with the additive effect of the two compounds alone, suggesting a synergistic effect. Lupeol treatment resulted in a significant inhibition in growth of tumors with concomitant reduction in prostate-specific antigen secretion in athymic nude mice implanted with CWR22Rnu1 cells. Because early clinical prostate cancer growth is an androgen-dependent response, the results of the present study suggest that Lupeol may have a potential to be an effective agent against prostate cancer.

"Isoflavones from both soy and Osage orange recently have been demonstrated by other laboratories to protect brain cells, or neurons, from the toxic effect of amyloid beta peptide. Amyloid beta peptide is believed to be responsible for the degeneration of neurons in Alzheimer's patients. However, the precise mechanisms by which isoflavones block the toxicity of amyloid beta peptide are unknown. Soybeans contain less than one-tenth of one percent isoflavone compounds, Whaley said. By comparison, between 5 percent and 10 percent of the dry mass of a bois d'arc apple is composed of isoflavone compounds. This plant has somehow developed the ability to produce extraordinary high levels of isoflavone compounds," Whaley said. "There is no known dietary plant that contains this high level of isoflavones.Whaley's research is focused on determining the molecular structure of the unique kind of isoflavones found in the Osage orange and on identifying the enzymes, and the genes that encode them, that are responsible for producing these compounds. Once the genes are identified and cloned, it may be possible to insert these genes into other, more palatable dietary plants, or forage plants eaten by livestock, he said.There are different kinds of isoflavones, each with their own unique structure," Whaley said. "The ones in the Osage orange have a peculiar structure that makes them very water insoluble. But they could be used as precursors in the chemical synthesis of other ingestible isoflavones.Whaley is particularly interested in how isoflavones protect brain cells from the toxic effects of the amyloid beta protein, which travels through the blood stream by hitching a ride within low-density lipoprotein, or LDL, particles. Whaley's recent research has focused on how isoflavones get inside, or partition into, lipid structures called micelles. These micelles are simple lipid structures that are chemically similar to LDL particles. To study the mechanisms for how isoflavones get inside the LDL particles in the body, Whaley has used micelles as a simple model. He adds a special detergent to water that forms micelles, and by adding isoflavones to the mix, he can then observe how the isoflavones position and orient themselves within the micelle structure. The position and orientation of isoflavones is a key consideration in understanding how they protect lipid structures from reactive oxygen species, which are also called free radicals. It is the reactivity of free radicals that ultimately is responsible for the aging process. "The next step is to take the techniques we've developed for studying detergent micelles and translate those techniques into more complex models and eventually into cells," he said. "The mechanism by which isoflavones block neurotoxicity could be related to their ability to lodge into lipid membranes, because that is where the precursor to amyloid beta peptide is usually located. Understanding the basic structure of the isoflavones and their interaction with lipid structures hopefully will give us insight into how natural chemicals in fruits and vegetables may lower the risk for developing diseases such as Alzheimer's disease or atherosclerosis. "Coming up with a magic bullet to treat Alzheimer's disease is a long way off, but I think there's a chance of learning something through natural products such as the isoflavones produced by Osage orange. They probably will not provide an immediate drug, but understanding their biological activities may indicate an avenue of research for developing a new pharmaceutical."

Abstract: The fruit of the osage orange tree (Maclua pomifera [Raf.] Schneider) was shown to contain at least four pigments with antioxidant activity. Pomiferin was present as 3 to 4% of the dry fruit and, as a primary antioxidant, was responsible for 20-25% of the activity exhibited by methanol extracts. An unidentified substance was also present which reacted synergistically with pomiferin, increasing its contribution to the overall activity to approximately 75%. The three new pigments, totalling 0.5% to .75% of the dry fruit, all showed antioxidant activity exceeding that of pomiferin when tested at equal concentrations. A chromatographic procedure, employing anhydrous magnesium sulfate, was developed and applied successfully in separating mixtures of several iso-flavone pigments.


In Osage orange, osajin and pomiferin account for between 5 and 10 percent of the dry mass of the fruit tissue. This is an extraordinary amount of isoflavone and is much higher than the isoflavone content of soybean (approximately 0.1% by mass). Osajin and pomiferin were first isolated and characterized by researchers at The Ohio State University in the late 1930s. It appears from this study that a 90/10, v/v, methanol/water solvent was used to extract these isoflavones. O sajin and pomiferin have low solubility in solvent mixtures containing small amounts of water.

Abstract: The objective of the present 15-day study was to evaluate the cardioprotective potential of flavonoid pomiferin isolated from the infructences of Maclura pomifera , Moraceae, against ischemia-reperfusion induced injury in rat hearts as a model of antioxidant-based composite therapy. Studies were performed with isolated, modifi ed Langendorff-perfused rat hearts and ischemia of heart was initiated by stopping the coronary flow for 30 min, followed by 60 min of reperfusion (14 ml min-1 ). Wistar rats were divided into three groups. The treated group received pomiferin (5 mg/kg/day in 0.5% Avicel); the placebo group received only 0.5% Avicel; the intact group was left without any applications. Biochemical indicators of oxidative damage, lipid peroxidation product malondialdehyde, antioxidant enzymes (superoxide dismutase, glutathione peroxidase, total antioxidant activity in serum and myocardium has been evaluated. We also examined the effect of pomiferin on cardiac function (left ventricular end-diastolic pressure, left ventricular pressure, peak positive +dP/dt (rate of pressure development) after ischemia and reperfusion. Our results demonstrate that pomiferin attenuates the myocardial dysfunction provoked by ischemiareperfusion. This was confirmed by the increase in both the antioxidant enzyme values and the total antioxidant activity. The cardio-protection provided by pomiferin treatment results from the suppression of oxidative stress and correlates with the improved ventricular function.

Abstract: Recent findings that many human chronic diseases are associated with oxidative stresses have instigated the search for dietary antioxidants. Many phytochemicals, particularly phenolic compounds, have been found to possess strong antioxidant activity and reduce the risks of those diseases. Isoflavones, a special phenolic group found in soybean, have been found to act as antioxidants in some model systems. This study investigated the isoflavone content in a unique nonedible tree fruit, Osage orange [Maclura pomifera (Raf.) Schneid], and methods for the extraction, identification, and quantification of the two major isoflavones, osajin and pomiferin, were developed. The ethyl acetate extract contained 25.7% osajin and 36.2% pomiferin, and the two isoflavones were at 9.5 g kg(-1) of fresh Osage orange. Two model systems, FRAP and beta-CLAMS, were used to measure the antioxidant activity of these two isoflavones. Pomiferin was found to be a strong antioxidant in both systems, comparable to the antioxidant vitamins C and E and the synthetic antioxidant BHT. Osajin and the two soybean isoflavones (genistein and daidzein) showed no antioxidant activity. Although the Osage orange fruit is not a food source, it is considered to be safe and, therefore, a potentially good source of an antioxidant nutraceutical and functional food ingredient.

The fruits of Maclura pomifera extracts inhibits glioma stem-like cell growth and invasion.
Zhao D1, Yao C, Chen X, Xia H, Zhang L, Liu H, Jiang X, Dai Y, Liu J.

Abstract
Glioma is the most common primary intracranial tumour. Recently, growing evidence showed that glioma possesses stem-like cells, which are thought to be chemo- and radio-resistant and believed to contribute to the poor clinical outcomes of these tumours. In this study, we found that stem-like glioma cells (CD133+) were significantly increased in neurosphere cells, which are highly invasive and resistant to multiple chemotherapeutic agents. From our natural products library, we screened 48 natural products and found one compound, Pomiferin, which was of particular interest. Our results showed that Pomiferin could inhibit cell viability, CD133+ cell population, sphere formation, and invasion ability of glioma neurosphere cells. We also found that multiple stemness-associated genes (BIM1, Nestin, and Nanog) were down-regulated by Pomiferin treatment of glioma neurosphere cells. Taken together, our results suggest that Pomiferin could kill the cancer stem-like cells in glioma and may serve as a potential therapeutic agent in the future.

Summary:


I had origninally concluded that the Osage Orange fruit was probably of debatable medicinal value when compared to the inner bark and leaf. However, I can see as you can from the above research that the fruit may offer some unique compounds not found in other parts of this tree. Neverheless, I have to conclude that this tree's inner bark probably will offer the best source for the complete "soup" of all the medicinal compounds that this tree has to offer. After all, the inner bark is the pathway by which the leaf, the fruit and the heartwood obtains their unique components and it is the inner bark that has long been esteemed in herbal medicine as the most valuable part of any medicinal tree. The leaf and the fruits are the easiest components to obtain for medicine making. The heartwood, less so and the inner bark probably the most time consuming of all to collect. Prices for Osage Orange tincture will reflect this. You will pay higher prices for the inner bark and heartwood tincture as compared to the fruit and leaf tinctures.


1) Osage Orange Leaf tincture will likely contain five valuable flavonoids and anti-fungal extracts similar to its close relative, Maclura tinctoria. A study has shown the leaf to contain the triterpene Lupine which has shown to have antimicrobial effects, along with deactivating melonoma cells and other cytotoxic activities. Its leaf should be of use as a cheap safe means of fighting melonoma, antimicrobial & fungal infections.

2) Osage Orange Heartwood tincturecontains the stilbenes, Oxyresveratrol & Tetrahydroxystilbene (THS), plus the the flavanonol, dihydromorin and like the leaf should be invaluable in fighting fungal infections, has antiviral potential, neuroprotective effects, plus as a general immune stimulant. It appears that oxyresveratrol may be more desirable on several counts than the highly touted resveratrol!

3)Osage Orange fruit tincture has been shown to contain the isoflavones:scandenone, osajin, pomiferin and auriculasin, along with the triterpenes lupeol, butyrospermol, and lupane-3B,20-diol. Lupeol has been linked to induction of apoptosis and inhibition of growth of pancreatic and prostate cancer cells. One would surmise this tincture would be of value as an anti-microbial, anit-tumor and anti-cancer medicinal along with a cell and cardio protectorate.

4) Osage Orange inner bark tinctures should traditionally prove to have the best combination of all of the previously described phytochemicals in one tincture. Almost without exception in the herbal literature, the inner bark (cambium layer and phloem) contains the most powerful concentration of a plant's chemicals and is the primary location harvested in creating a medicinal extract.

For more in-depth information on the Osage Orange as a medicinal plant and how to process this plant in a home or clinic, purchase any of my three books. Click on the book images to the right to be taken to Amazon.